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Understanding the patterns of treatment response is critical for the treatment of patients with schizophrenia; one way to achieve this is through using a longitudinal dynamic process study design.
This study aims to explore the response trajectory of antipsychotics and compare the treatment responses of seven different antipsychotics over 6 weeks in patients with schizoprenia (trial registration: Chinese Clinical Trials Registry Identifier: ChiCTR-TRC-10000934).
Data were collected from a multicentre, randomised open-label clinical trial. Patients were evaluated with the Positive and Negative Syndrome Scale (PANSS) at baseline and follow-up at weeks 2, 4 and 6. Trajectory groups were classified by the method of k-means cluster modelling for longitudinal data. Trajectory analyses were also employed for the seven antipsychotic groups.
The early treatment response trajectories were classified into a high-trajectory group of better responders and a low-trajectory group of worse responders. The results of trajectory analysis showed differences compared with the classification method characterised by a 50% reduction in PANSS scores at week 6. A total of 349 patients were inconsistently grouped by the two methods, with a significant difference in the composition ratio of treatment response groups using these two methods (χ2 = 43.37, P < 0.001). There was no differential contribution of high- and low trajectories to different drugs (χ2 = 12.52, P = 0.051); olanzapine and risperidone, which had a larger proportion in the >50% reduction at week 6, performed better than aripiprazole, quetiapine, ziprasidone and perphenazine.
The trajectory analysis of treatment response to schizophrenia revealed two distinct trajectories. Comparing the treatment responses to different antipsychotics through longitudinal analysis may offer a new perspective for evaluating antipsychotics.
To reveal the thermal shock resistance of double-layer thermal barrier coatings (TBCs), two types of TBCs were prepared via atmospheric plasma spraying, i.e., Gd2Zr2O7/yttria-stabilized zirconia (GZ/YSZ) TBCs and La2Zr2O7 (LZ)/YSZ TBCs, respectively. Subsequently, thermal cycling tests of the two TBCs were conducted at 1100 °C and their thermal shock resistance and failure mechanism were comparatively investigated through experiments and the finite element method. The results showed that the thermal shock failure of the two TBCs occurred inside the top ceramic coating. However, the GZ/YSZ TBCs had longer thermal cycling life. It was the mechanical properties of the top ceramic coating, and the thermal stresses arising from the thermal mismatch between the top ceramic coating and the substrate that determined the thermal cycling life of the two TBCs together. Compared with the LZ layer in the LZ/YSZ TBCs, the GZ layer in the GZ/YSZ TBCs had smaller elastic modulus, larger fracture toughness, and smaller thermal stresses, which led to the higher crack propagation resistance and less spallation tendency of the GZ/YSZ TBCs. Therefore, the GZ/YSZ TBCs exhibited superior thermal shock resistance to the LZ/YSZ TBCs.
Isolated congenital tricuspid regurgitation other than Ebstein’s anomaly was rare especially for children. The objective of this study was to investigate the clinical characteristics and to assess the results of tricuspid valvuloplasty for children with isolated tricuspid regurgitation.
From January 2010 to June 2019, 10 consecutive patients with isolated tricuspid regurgitation who were unresponsive to drug therapy underwent tricuspid valvuloplasty in our hospital. Patients’ clinical data were analysed retrospectively.
Mean age at operation was 48.5 ± 31.0 (range: 9–106) months and mean weight at operation was 16.1 ± 6.9 (range: 8.6–33.0) kg. All patients presented severe isolated tricuspid regurgitation. According to pathological lesions, the main causes accounted for chordae tendinea rupture (3/10), leaflet cleft (2/10), mal-connected chordal tendinea to leaflets (2/10), elongated chordae (1/10) and chordae absent (1/10), and severe anterior leaflet dysplasia (1/10). Individualised tricuspid valvuloplasty was adapted to all of them successfully. Post-operative echocardiography showed no tricuspid regurgitation in two patients and mild regurgitation in eight patients. The cardiothoracic ratios on their chest roentgenograms decreased from 0.59 ± 0.05 to 0.54 ± 0.05. At the latest follow-up (50.4 ± 47.2 months), echocardiography showed that mild to moderate tricuspid regurgitation in seven patients, moderate tricuspid regurgitation in three patients, and no patient with severe tricuspid regurgitation. All patients were in NYHA functional class I.
For patients with isolated tricuspid regurgitation who were not well responsive to drug therapy, individualised tricuspid valve repair can achieve an excellent result.
Dolostones are widely developed in the middle Permian rocks of East Yunnan, China, mainly in the shoal-facies Maokou Formation. The previously reported dolostone formation mechanisms cannot explain the distribution and geochemical characteristics of these dolostones, in particular their strontium, magnesium and oxygen isotope signatures. To help predict the distribution of dolostone reservoirs and reduce the exploration risk and cost, this study proposes a new model of dolomitization: open thermal convection dolomitization. In this new dolomitization model, Mg2+ in dolomitizing fluids originates mostly from seawater, with a minor component coming from deep hydrothermal fluids. Elevated heat flux (in this case due to the nearby Emei mantle plume) causes spatial temperature variations in the fluid along the circulation flow pathways, resulting in fast and pervasive dolomitization of limestone. The proposed model not only explains the characteristics and distribution of dolostones in the study area but also serves as a reference for predicting the distribution of dolostones in other areas subjected to thermal convection.
The surface topology of biomaterial has a definite effect on the growth behavior of nerve cells for peripheral nerve regeneration. In this study, the silk fibroin (SF) film with different anisotropic microgroove/ridge was constructed by micropatterning technology. The effects of topologies width on the directional growth of dorsal root ganglion (DRG) neurons were evaluated. The results showed that the topological structure of the SF film with higher SF concentration was more clear and complete. The microtopography of the SF film with a concentration of 15% and a groove width of around 30 μm could effectively guide the directional growth of the nerve fibers of DRG. And nerve fibers could obviously form nerve fiber bundles which may have a certain pavement effect on the recovery of nerve function. The study indicated that the SF film with a specific width of the topological structure may have potential applications in the field of directional nerve regeneration.
The Arsenic (+3 oxidation state) methyltransferase (AS3MT) gene has been identified as a top risk gene for schizophrenia in several large-scale genome-wide association studies. A variable number tandem repeat (VNTR) of this gene is the most significant expression quantitative trait locus, but its role in brain activity in vivo is still unknown.
We first performed a functional magnetic resonance imaging (fMRI) scan of 101 healthy subjects during a memory span task, trained all subjects on an adaptive memory span task for 1 month, and finally performed another fMRI scan after the training. After excluding subjects with excessive head movements for one or more scanning sessions, data from 93 subjects were included in the final analyses.
The VNTR was significantly associated with both baseline brain activation and training-induced changes in multiple regions including the prefrontal cortex and the anterior and posterior cingulate cortex. Additionally, it was associated with baseline brain activation in the striatum and the parietal cortex. All these results were corrected based on the family-wise error rate method across the whole brain at the peak level.
This study sheds light on the role of AS3MT gene variants in neural plasticity related to memory span training.
Primary liver cancer is the third leading cause of cancer-related death worldwide. Most patients are diagnosed at late stages with poor prognosis; thus, identification of modifiable risk factors for primary prevention of liver cancer is urgently needed. The well-established risk factors of liver cancer include chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV), heavy alcohol consumption, metabolic diseases such as obesity and diabetes, and aflatoxin exposure. However, a large proportion of cancer cases worldwide cannot be explained by current known risk factors. Dietary factors have been suspected as important, but dietary aetiology of liver cancer remains poorly understood. In this review, we summarised and evaluated the observational studies of diet including single nutrients, food and food groups, as well as dietary patterns with the risk of developing liver cancer. Although there are large knowledge gaps between diet and liver cancer risk, current epidemiological evidence supports an important role of diet in liver cancer development. For example, exposure to aflatoxin, heavy alcohol drinking and possibly dairy product (not including yogurt) intake increase, while intake of coffee, fish and tea, light-to-moderate alcohol drinking and several healthy dietary patterns (e.g. Alternative Healthy Eating Index) may decrease liver cancer risk. Future studies with large sample size and accurate diet measurement are warranted and need to consider issues such as the possible aetiological heterogeneity between liver cancer subtypes, the influence of chronic HBV or HCV infection, the high-risk populations (e.g. cirrhosis) and a potential interplay with host gut microbiota or genetic variations.
Findings for the roles of dairy products, Ca and vitamin D on ovarian cancer risk remain controversial. We aimed to assess these associations by using an updated meta-analysis. Five electronic databases (e.g. PubMed and Embase) were searched from inception to 24 December 2019. Pooled relative risks (RR) with 95 % CI were calculated. A total of twenty-nine case–control or cohort studies were included. For comparisons of the highest v. lowest intakes, higher whole milk intake was associated with increased ovarian cancer risk (RR 1·35; 95 % CI 1·15, 1·59), whereas decreased risks were observed for higher intakes of low-fat milk (RR 0·84; 95 % CI 0·73, 0·96), dietary Ca (RR 0·71; 95 % CI 0·60, 0·84) and dietary vitamin D (RR 0·80; 95 % CI 0·67, 0·95). Additionally, for every 100 g/d increment, increased ovarian cancer risks were found for total dairy products (RR 1·03; 95 % CI 1·01, 1·04) and for whole milk (RR 1·07; 95 % CI 1·03, 1·11); however, decreased risks were found for 100 g/d increased intakes of low-fat milk (RR 0·95; 95 % CI 0·91, 0·99), cheese (RR 0·87; 95 % CI 0·76, 0·98), dietary Ca (RR 0·96; 95 % CI 0·95, 0·98), total Ca (RR 0·98; 95 % CI 0·97, 0·99), dietary vitamin D (RR 0·92; 95 % CI 0·87, 0·97) and increased levels of circulating vitamin D (RR 0·84; 95 % CI 0·72, 0·97). These results show that whole milk intake might contribute to a higher ovarian cancer risk, whereas low-fat milk, dietary Ca and dietary vitamin D might reduce the risk.
The aim of this study was to investigate the combined effect of n-3 fatty acids (EPA and DHA, at an EPA:DHA ratio of 150:500) and phytosterol esters (PS) on non-alcoholic fatty liver disease (NAFLD) patients. We conducted a randomised, double-blind, placebo-controlled trial. Ninety-six NAFLD subjects were randomly assigned to the following groups: the PS group (receiving 3·3 g/d PS); the FO group (receiving 450 mg EPA + 1500 mg DHA/d); the PS + FO combination group (receiving 3·3 g/d PS and 450 mg EPA + 1500 mg DHA/d) and the PO group (a placebo group). The baseline clinical characteristics of the four groups were similar. The primary outcome was liver:spleen attenuation ratio (L:S ratio). The percentage increase in liver–spleen attenuation (≤1) in the PS + FO group was 36 % (P = 0·083), higher than those in the other three groups (PS group, 11 %, P = 0·519; FO group, 18 %, P = 0·071; PO group, 15 %, P = 0·436). Compared with baseline, transforming growth factor-β (TGF-β) was significantly decreased in the three study groups at the end of the trial (PS, P = 0·000; FO, P = 0·002; PS + FO, P = 0·001) and TNF-α was significantly decreased in the FO group (P = 0·036), PS + FO group (P = 0·005) and PO group (P = 0·032) at the end of the intervention. Notably, TGF-β was reduced significantly more in the PS + FO group than in the PO group (P = 0·032). The TAG and total cholesterol levels of the PS + FO group were reduced by 11·57 and 9·55 %, respectively. In conclusion, co-supplementation of PS and EPA + DHA could increase the effectiveness of treatment for hepatic steatosis.
Maintenance haemodialysis (MHD) is the use of a machine to filter wastes, salts and fluid from blood for at least 3 months to prolong the life of patients with advanced kidney failure. Although low dietary energy intake (DEI) has been observed in MHD patients, few studies have related DEI to the risk of mortality. To explore this relationship, a study included 1039 MHD patients from eight centres was conducted. DEI was assessed by three 24-h diet recalls and was normalised to ideal body weight (IBW). All-cause mortality and CVD mortality were the primary and secondary outcomes, respectively. During a median follow-up of 28 months, a U-shaped relationship was observed between DEI and all-cause or CVD mortality. The risk of all-cause mortality decreased significantly with the increase of DEI in participants with DEI <167·4 kJ/kg IBW per d (hazard ratio (HR) 0·98; 95 % CI 0·96, 1·00) and increased significantly with the increase of DEI in those with DEI ≥167·4 kJ/kg IBW per d (HR 1·12; 95 % CI 1·04, 1·20). Similarly, the risk of CVD mortality decreased with the increase of DEI in participants with DEI <152·7 kJ/kg IBW per d (HR 0·96; 95 % CI 0·93, 0·99) and increased with the increase of DEI in participants with DEI ≥152·7 kJ/kg IBW per d (HR 1·11; 95 % CI 1·04, 1·18). In summary, there was a U-shaped association between DEI and all-cause or CVD mortality, with a turning point at about 167·4 and 152·7 kJ/kg IBW per d, respectively, in MHD patients.
The microbiota–gut–brain axis, especially the microbial tryptophan (Trp) biosynthesis and metabolism pathway (MiTBamp), may play a critical role in the pathogenesis of major depressive disorder (MDD). However, studies on the MiTBamp in MDD are lacking. The aim of the present study was to analyze the gut microbiota composition and the MiTBamp in MDD patients.
We performed shotgun metagenomic sequencing of stool samples from 26 MDD patients and 29 healthy controls (HCs). In addition to the microbiota community and the MiTBamp analyses, we also built a classification based on the Random Forests (RF) and Boruta algorithm to identify the gut microbiota as biomarkers for MDD.
The Bacteroidetes abundance was strongly reduced whereas that of Actinobacteria was significantly increased in the MDD patients compared with the abundance in the HCs. Most noteworthy, the MDD patients had increased levels of Bifidobacterium, which is commonly used as a probiotic. Four Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologies (KOs) (K01817, K11358, K01626, K01667) abundances in the MiTBamp were significantly lower in the MDD group. Furthermore, we found a negative correlation between the K01626 abundance and the HAMD scores in the MDD group. Finally, RF classification at the genus level can achieve an area under the receiver operating characteristic curve of 0.890.
The present findings enabled a better understanding of the changes in gut microbiota and the related Trp pathway in MDD. Alterations of the gut microbiota may have the potential as biomarkers for distinguishing MDD patients form HCs.
Echinococcus granulosus sensu stricto (s.s.), Echinococcus multilocularis and Echinococcus canadensis are the common causes of human echinococcosis in China. An accurate species identification tool for human echinococcosis is needed as the treatments and prognosis are different among species. The present work demonstrates a method for the simultaneous detection of these three Echinococcus species based on multiplex polymerase chain reaction (mPCR). Specific primers of this mPCR were designed based on the mitochondrial genes and determined by extensive tests. The method can successfully detect either separated or mixed target species, and generate expected amplicons of distinct size for each species. Sensitivity of the method was tested by serially diluted DNA, showing a detection threshold as less as 0.32 pg for both E. granulosus s.s. and E. canadensis, and 1.6 pg for E. multilocularis. Specificity assessed against 18 other parasites was found to be 100% except weakly cross-react with E. shiquicus. The assay was additionally applied to 69 echinococcosis patients and 38 healthy persons, confirming the high reliability of the method. Thus, the mPCR described here has high application potential for clinical identification purposes, and can further provide a useful tool for evaluation of serology and imaging method.
The East Kunlun Orogen (EKO) is the NW part of the Central China Orogenic Belt, which records the evolutionary history of the Proto- and Palaeo-Tethys Oceans from the Cambrian to the Triassic. An Early Palaeozoic eclogite belt has been recognized in recent years, which extends discontinuously for ∼500 km as three eclogite-bearing terranes. In this study, we report an integrated study of zircon grains from mica-schists accompanying the eclogites, in terms of mineral inclusions, U–Pb age systematics and P–T conditions. The presence of coesite is identified, as inclusions within the metamorphic domain of zircons, which provides unambiguous evidence for subducted terrigenous clastic rocks of the Proto-Tethys Ocean exhumed from coesite-forming depths. U–Pb dating of the metamorphic zircons yields a concordia age of 426.5 ± 0.88 Ma, which is likely to be the time of ultrahigh-pressure metamorphism in the Kehete terrane. P–T calculations suggest that metapelite may have experienced a clockwise P–T path with peak P/T conditions of 685 ± 41 °C and >28 kbar, and equilibrated at 482–566 °C and 5.6–8.9 kbar during subsequent exhumation. The high-pressure – ultrahigh-pressure (HP-UHP) metamorphic belt within the EKO may have formed by collision between the Qaidam Block and the South Kunlun Block, as a consequence of the closure of the Proto-Tethys Ocean.
A compact circularly polarized (CP) rectenna with low profile and high efficiency based on the artificial magnetic conductor (AMC) is proposed in this paper. The receiving CP antenna is a coplanar stripline fed dual rhombic loop with an AMC reflector. The proposed AMC reflector not only improves the antenna gain to 9.8 dBi but also decreases the profile to 0.1 λ0. The AMC reflector also makes the antenna have a harmonic suppression function so the low pass filter between the rectifying circuit and the antenna could be omitted and the rectenna has a compact structure. According to the measured results, the rectenna has the highest conversion efficiency of 76% on the load of 240 Ω with the received power of 117.5 mW. When the linearly polarized transmitting antenna is rotated, the conversion efficiency of the CP rectenna maintains a constant high conversion efficiency of 74%. The compact structure and CP operation of the rectenna made it a good candidate of the wireless battery for some electronic devices and far-distance microwave power transmission.
Underground Nuclear Astrophysics in China (JUNA) will take the advantage of the ultra-low background in Jinping underground lab. High current accelerator with an ECR source and detectors were commissioned. JUNA plans to study directly a number of nuclear reactions important to hydrostatic stellar evolution at their relevant stellar energies. At the first period, JUNA aims at the direct measurements of 25Mg(p,γ)26 Al, 19F(p,α) 16 O, 13C(α, n) 16O and 12C(α,γ) 16O near the Gamow window. The current progress of JUNA will be given.
Living Caprini are dominant bovids in the pan-Tibetan area that are strongly adapted to dry steppe and high-mountain meadow habitats. Some taxa with Holarctic distributions, e.g., Ovis Linnaeus, 1758, were thought to originate on the Tibetan Plateau and subsequently dispersed elsewhere, which was depicted as an ‘out of Tibet’ story. However, except for some information on a stem caprine assemblage from the Qaidam Basin, the early evolution of Caprini around the Tibetan Plateau is poorly known. Here, we report new material of Olonbulukia tsaidamensis Bohlin, 1937, which was a member of this stem caprine assemblage, from the Wuzhong region, northern China, confirming the similarity of the Wuzhong Fauna and ‘Qaidam Fauna.’ Based on a biometric study of horncores from the ‘Qaidam’ and Wuzhong faunas, we recognize six taxa from this stem caprine assemblage: O. tsaidamensis, O. sp., Qurliqnoria cheni Bohlin, 1937, Tossunnoria pseudibex Bohlin, 1937, ?Protoryx cf. P. enanus Köhler, 1987, and cf. Pachytragus sp. Among these taxa, Q. cheni and T. pseudibex are probably related to some extant Tibetan endemic species, e.g., the Tibetan antelope, Pantholops hodgsonii (Abel, 1826), and the Himalayan tahr, Hemitragus jemlahicus (Smith, 1826). Others might be ancestral to the Turolian caprine assemblages and even possibly gave rise to the extant Caprina. This work reveals an early radiation of stem caprines along the northern side of the rising Tibetan Plateau and indicates a mixed pattern of pan-Tibetan stem caprine evolution prior to their dispersal out of the Tibetan Plateau.
Healthcare-associated infections (HAIs) are a major worldwide public-health problem, but less data are available on the long-term trends of HAIs and antimicrobial use in Eastern China. This study describes the prevalence and long-term trends of HAIs and antimicrobial use in a tertiary care teaching hospital in Hefei, Anhui, China from 2010 to 2017 based on annual point-prevalence surveys. A total of 12 505 inpatients were included; 600 HAIs were recorded in 533 patients, with an overall prevalence of 4.26% and a frequency of 4.80%. No evidence was found for an increasing or decreasing trend in prevalence of HAI over 8 years (trend χ2 = 2.15, P = 0.143). However, significant differences in prevalence of HAI were evident between the surveys (χ2 = 21.14, P < 0.001). The intensive care unit had the highest frequency of HAIs (24.36%) and respiratory tract infections accounted for 62.50% of all cases; Escherichia coli was the most common pathogen (16.67%). A 44.13% prevalence of antimicrobial use with a gradually decreasing trend over time was recorded. More attention should be paid to potential high-risk clinical departments and HAI types with further enhancement of rational antimicrobial use.
Previous studies have indicated that some food items and nutrients are associated with uric acid metabolism in humans. However, little is known about the role of dietary patterns in hyperuricaemia. We designed this case–control study to evaluate the associations between dietary patterns and newly diagnosed hyperuricaemia in Chinese adults. A total of 1422 cases and 1422 controls were generated from 14 538 participants using the 1:1 ratio propensity score matching methods. Dietary intake was assessed using a validated self-administered FFQ. Dietary patterns were derived by factor analysis. Hyperuricaemia was defined as concentrations of serum uric acid higher than 7 mg/dl (416·5 μmol/l) for men and 6 mg/dl (357 μmol/l) for women. Three dietary patterns were derived by factor analysis: sweet pattern; vegetable pattern; animal foods pattern. The animal foods pattern characterised by higher intake of an animal organ, seafood and processed meat products was associated with higher prevalence of newly diagnosed hyperuricaemia (Pfor trend<0·01) after adjustment. Compared with the participants in the lowest quartile of the animal foods pattern, the OR of newly diagnosed hyperuricaemia in the highest quartile was 1·50 (95 % CI 1·20, 1·87). The other two dietary patterns were not associated with the prevalence of newly diagnosed hyperuricaemia after adjustment. In conclusion, a diet rich in animal organ, seafood and processed meat products is associated with higher prevalence of newly diagnosed hyperuricaemia in a Chinese population. Further cohort studies and randomised controlled trials are required to clarify these findings.
In this paper we present new petrological and whole-rock geochemical data for the Palaeoproterozoic metasedimentary rocks in the upper part of the Ji'an Group within the Jiao–Liao–Ji Belt, China, as well as zircon U–Pb age dates and in situ Lu–Hf isotope data. The new data improve our understanding of the original nature of the metasedimentary rocks, further providing insights into their tectonic setting and the evolutionary history of the northern segment of the Jiao–Liao–Ji Belt. The zircons can be divided into two groups, viz., one of magmatic origin and the other of metamorphic origin. Zircon U–Pb dating gave mean or statistical peak ages for the magmatic zircons at 2035, 2082, 2178, 2343–2421, 2451–2545, 2643–2814 and 2923–3446 Ma, and mean peak ages for the metamorphic zircons at 1855 and 1912 Ma, which indicate a maximum depositional age of 2.03 Ga and two-stage metamorphic events at c. 1.91 and 1.85 Ga for the metasedimentary rocks. Geochemical data show that (1) the protoliths of these rocks were mainly sandstones, greywackes and claystones, together with some shales; (2) the main sources of the sedimentary material were Palaeoproterozoic granites and acid volcanic rocks, with minor contributions from Archaean granitic rocks; and (3) the sediments were deposited in an active continental margin setting. Moreover, along the northeastern margin of the Eastern Block of the North China Craton there is evidence of ancient crustal materials as old as 3.76 Ga, and multiple crustal growth events at 3.23–3.05, 2.80–2.65, 2.54–2.45 and 2.28–2.08 Ga.
Disclosing the diagnosis of Alzheimer's disease (AD) to a patient is controversial. There is significant stigma associated with a diagnosis of AD or dementia in China, but the attitude of the society toward disclosure of such a diagnosis had not been formally evaluated prior to our study. Therefore, we aimed to evaluate the attitude toward disclosing an AD diagnosis to patients in China with cognitive impairment from their caregivers, and the factors that may affect their attitude.
We designed a 17-item questionnaire and administered this questionnaire to caregivers, who accompanied patients with cognitive impairment or dementia in three major hospitals in Shanghai, China. The caregiver's attitude toward disclosing the diagnosis of AD as evaluated by the questionnaire was compared to that of disclosing the diagnosis of terminal cancer.
A majority (95.7%) of the 175 interviewed participants (mean 14.2 years of education received) wished to know their own diagnosis if they were diagnosed with AD, and 97.6% preferred the doctor to tell their family members if they were diagnosed with AD. If a family member of the participants suffered from AD, 82.9% preferred to have the diagnosis disclosed to the patient. “Cognitive impairment” was the most accepted term by caregivers to disclose AD diagnosis in Chinese.
This study suggests most of the well-educated individuals in a Chinese urban area favored disclosing the diagnosis when they or their family members were diagnosed with AD.