Objective: To analyze the mediating effects of loneliness and depressive symptoms on family functioning and life satisfaction among rural elderly patients with chronic diseases, so as to provide the basis for improving the life satisfaction of this population. Methods: Rural elderly patients with chronic diseases aged ≥60 years in Qiannan Buyi and Miao Autonomous Prefecture, Guizhou Province were selected using a multi-stage stratified random cluster sampling method from June to September 2022. Basic information such as gender, age, and chronic diseases were collected. Family function, life satisfaction, loneliness and depressive symptoms were evaluated using Family Care Index Scale, the Satisfaction with Life Scale, the b-item Revised VCLA Loneliness Sale and the 15-item Geriatric Depression Scale, respectively. The structural equation model was constructed using Amos software to analyze the mediating effects of loneliness and depressive symptoms on the relationship between family function and life satisfaction. The Bootstrap method was employed to test the mediating effects. Results: A total of 1 145 rural elderly patients with chronic diseases were recruited, including 517 males (45.15%) and 628 females (54.85%). Among the participants, 657 individuals (57.38%) were aged 60-<71 years, and 540 individuals (47.16%) had three or more chronic diseases. The scores for family function, life satisfaction, loneliness, and depressive symptoms were (3.90±1.18), (18.88±5.25), (12.88±2.99), and (6.65±2.26), respectively. Mediating effect analysis showed that family function had a direct positive effect on life satisfaction (β=0.179, 95%CI: 0.126-0.231). It also indirectly positively influenced the life satisfaction of rural elderly patients with chronic diseases through the independent mediating effect of depressive symptoms (β=0.035, 95%CI: 0.021-0.054) and the chained mediating effect of loneliness and depressive symptoms (β=0.021, 95%CI: 0.013-0.030). The mediating effect of depressive symptoms accounted for 14.89% of the total effect, while the chained mediating effect of loneliness and depressive symptoms accounted for 8.94% of the total effect. Conclusion Good family function can directly enhance the life satisfaction of rural elderly patients with chronic diseases and can also indirectly improve their life satisfaction by reducing loneliness and depressive symptoms.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by the highest mortality among carcinomas. The pathogenesis of PDAC requires elevated autophagy, inhibition of which using hydroxychloroquine has shown promise. However, current realization is impeded by its suboptimal use and unpredictable toxicity. Attempts to identify novel autophagy-modulating agents from already approved drugs offer a rapid and accessible approach. Here, using a patient-derived organoid model, we performed a comparative analysis of therapeutic responses among various antimalarial/fungal/parasitic/viral agents, through which econazole (ECON), an antifungal compound, emerged as the top candidate. Further testing in cell-line and xenograft models of PDAC validated this activity, which occurred as a direct consequence of dysfunctional autophagy. More specifically, ECON boosted autophagy initiation but blocked lysosome biogenesis. RNA sequencing analysis revealed that this autophagic induction was largely attributed to the altered expression of activation transcription factor 3 (ATF3). Increased nuclear import of ATF3 and its transcriptional repression of inhibitor of differentiation-1 (ID-1) led to inactivation of the AKT/mammalian target of rapamycin (mTOR) pathway, thus giving rise to autophagosome accumulation in PDAC cells. The magnitude of the increase in autophagosomes was sufficient to elicit ER stress-mediated apoptosis. Furthermore, ECON, as an autophagy inhibitor, exhibited synergistic effects with trametinib on PDAC. This study provides direct preclinical and experimental evidence for the therapeutic efficacy of ECON in PDAC treatment and reveals a mechanism whereby ECON inhibits PDAC growth.

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.

Objective: To investigate the contamination status of Salmonella, Vibrio parahaemolyticus and Staphylococcus aureus and bacterial resistance in retail meat products in Taiyuan, Shanxi Province. Methods: In the epidemic season of diarrhea in 2017, poultry and meat product specimens were randomly collected from the farmer′s markets and supermarkets of 10 districts and counties of Taiyuan. Salmonella, Vibrio parahaemolyticus and Staphylococcus aureus were isolated form these specimens. Serotypes of Salmonella strains were analyzed. ELSIA was used to detect Staphylococcus aureus enterotoxin (A-E). Vibrio parahaemolyticus isolates were tested for the virulence genes encoding direct hemolysin (tdh) and indirect hemolysin (trh). Antibiotic resistance of the three food-borne pathogens were analyzed using microdilution methods. Results: A total of 38 food-borne pathogens were isolated from 123 poultry and livestock meat product specimens with a positive rate of 30.9%, of which mainly were Salmonella (26 strains, 21.1%), followed by Vibrio parahaemolyticus (8 strains, 6.5%) and Staphylococcus aureus (4 strains, 3.3%). The 26 strains of Salmonella belonged to 10 serotypes. The Salmonella strains isolated from pork specimens had diverse serotypes. Salmonella serovar Derby, Salmonella serovar Gold-coast and Salmonella serovar Liverpool were isolated from raw and cooked pork food for the first time in Taiyuan. All Salmonella strains isolated form chicken products were Salmonella enteritis. The enterotoxin types of the four Staphylococcus aureus strains were three E-type and one complex type (A/E). All Vibrio parahaemolyticus isolates were negative for tdh or trh gene. Ampicillin, chloramphenicol, streptomycin, sulphonamides and tetracyclines (ACSSuT) resistance was prevalent in multi-drug resistant (MDR) Salmonella strains, but there was high sensitivity to fluoroquinolones and cephalosporins. MDR Staphylococcus aureus accounted for 75%. No third-generation cephalosporin- or fluoroquinolone-resistant or MDR Vibrio parahaemolyticus strains were isolated. Conclusions There were food-borne multi-pathogenic bacteria contamination in retail raw and cooked meat products in Taiyuan. Salmonella strains had diverse serotypes and high MDR rate. It was suggested that the regulatory authorities should strengthen the management of antibiotic use in aquaculture and specialized laboratory-based monitoring of meat supply chain.

OBJECTIVETo observe the effect difference between acupuncture of "five needles therapy" and conventional acupuncture for asthma of latent cold phlegm-fluid in the lung.

METHODSTwo hundred and ten cases were randomly assigned into an observation group and a control group, 105 cases in each one. Finally 7 cases were dropped out in the observation group; 6 cases in the control group. Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12) were used in the observation group; conventional acupuncture was used in the control group, and the main acupoints were Feishu (BL 13), Zhongfu (LU 1), Tiantu (CV 22), Danzhong (CV 17), Kongzui (LU 6), Dingchuan (EX-B 1), Fenglong (ST 40), Fengmen (BL 12), Taiyuan (LU 9). The needles were retained for 30 min each time, once a day for continuous 12 days. The scores of the individual symptoms and signs were observed before treatment and on the 3rd, 6th, 9th, 12th days, including pant, cough, cough up phlegm, fullness and oppression in the chest and diaphragm, wheezing rale and shortness of breath. The clinical effects were compared between the two groups.

RESULTSThe scores of six individual symptoms and signs on the 3rd, 6th, 9th, and 12th days in the two groups were lower than those before treatment (all<0.05), except the score of wheezing rale in the control group on the 3rd day (>0.05). The scores of pant, wheezing rale, cough on the 3rd, 6th, 9th, and 12th days in the observation group were lower than those in the control group (all<0.05), except the score of wheezing rale score on the 3rd day (>0.05). There were no significant difference between the two groups about the scores of cough up phlegm, fullness and oppression in the chest and diaphragm and shortness of breath on the 3rd, 6th, 9th, and 12th days (all>0.05), except the score of fullness and oppression in the chest and diaphragm in the observation group was lower than that in the control group on the 12th day (<0.05). 46 cases were clinical cured, 39 cases were markedly effective, 10 cases were effective and 3 cases were ineffective in the observation group with the total effective rate of 96.9%. 23 cases were clinical cured, 43 cases were markedly effective, 24 cases were effective and 9 cases were ineffective in the control group with the total effective rate of 90.9%. The difference was statistical (<0.05).

CONCLUSION"Five needles therapy" has significant therapeutic effect for asthma of latent cold phlegm-fluid in the lung, which is better than conventional acupuncture.

Objective: o investigate the features of pathogenic bacteria for community-acquired bloodstream infection due to Gram-negative bacilli in patients with liver cirrhosis and optimal therapeutic strategy. Methods: A retrospective analysis was performed for the clinical data of patients with liver cirrhosis who were admitted to 302 Hospital of PLA due to community-acquired bloodstream infection from January 2010 to December 2015, and a statistical analysis was performed for their clinical features, pathogenic bacteria, and results of drug sensitivity test. The Pearson chi-square test was used for comparison of rates, and the Wilcoxon rank sum test was used for comparison of ranked data. Results: A total of 240 patients (including 178 male patients) with liver cirrhosis caused by various reasons were enrolled, with a mean age of 51.7 ± 11.1 years, an overall clinical remission rate of 80.42%, and an ineffective/mortality rate of 19.58%. The patients who used sensitive antibiotics within 12 hours after the onset of community-acquired bloodstream infection achieved a significantly higher improvement rate than those who used such drugs at more than 12 hours after onset (88.2% vs 58.1%, P < 0.001). The improvement rate achieved by the application of sensitive antibiotics at more than 12 hours after onset decreased with the increase in the Child-Pugh grade (P < 0.05). A total of 245 strains of Gram-negative bacilli were isolated, among which the six most common ones were 135 strains of Escherichia coli (55.1%), 62 strains of Klebsiella pneumoniae (25.3%), 16 strains of Aeromonas (6.5%), 4 strains of non-typhoidal Salmonella (1.6%), 3 strains of Enterobacter cloacae (1.2%), and 2 strains of Acinetobacter baumannii (0.8%). These Gram-negative bacilli had the highest sensitivity to meropenem (98.5%), followed by imipenem (97.9%), amikacin (97.5%), piperacillin/tazobactam (94.7%), cefmetazole (93.7%), and cefoperazone/sulbactam (93%). Different bacteria had different sensitivities to antibiotics. Conclusion Once community-acquired bloodstream infection occurs in patients with liver cirrhosis, highly sensitive antibiotics should be used as early as possible. Cefoperazone/sulbactam, piperacillin/tazobactam, imipenem, and meropenem can be used as first-line empirical antibiotics, and drug combination should be considered when necessary.

OBJECTIVETo investigate the accuracy and consistency of the detection of BCR-ABL tyrosine kinase domain point mutation among different laboratories.

METHODSEvery one of 6 laboratories prepared 10 cDNA samples from tyrosine kinase inhibitors resistant BCR-ABL (P210 or P190) positive patients'bone marrow or peripheral blood. Each cDNA sample was divided into 6 aliquots and delivered to the laboratories. All 6 laboratories tested BCR-ABL point mutations of 60 samples according to their own protocols. Peking University People's Hospital analyzed the comparison results based on both the reports and sequencing chromatogram from all laboratories.

RESULTSAll laboratories reported the same nucleotide and corresponding amino acid mutations in 37 samples (61.7%）. Of 60 samples, 53 had confirmed mutation types, and a total of 23 types were included; 1 had no mutation; mutation types of 6 samples could not be determined because of the big differences among chromatograms from different laboratories. Low percentages of mutants were significantly related to results inconsistency (P=0.008). Inconsistent result of one sample was caused by the unique chromatogram of the mutant L248V, and one by the non-coverage amplification of PCR product from different laboratories. Amplification was failed in 3 samples. Testing or sequencing mistakes occurred in 7 samples. The differences in the mutant percentages among laboratories were less than 20% in the 80.6% of samples with confirmed results. Low internal control gene copies (ABL<10 000) were significantly related to both failed amplification and big differences among chromatograms from different laboratories (P=0.005 and <0.001, respectively).

CONCLUSIONProblems in the clinical routine detection of BCR-ABL point mutation could be exposed and improvement could be achieved by sample exchange and comparison. Low percentage of mutant is the main reason which causes the discrepancy of BCR-ABL point mutation results among different laboratories.

Bone Marrow ; DNA Mutational Analysis ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Point Mutation ; Polymerase Chain Reaction

OBJECTIVETo revalidate the conversion factor（CF）for the conversion of BCR-ABL （P210）transcript levels to the international scale（BCR- ABLIS）in chronic myeloid leukemia（CML） which validated before.

METHODSPeking University People's Hospital（PKUPH）prepared the exchange samples for revalidation of CFs of 15 laboratories which validated nine or eighteen months ago. The fresh BCR-ABL（P210）（+）bone morrow or peripheral blood nucleated cells were diluted with BCR-ABL （P210）（-）cells to achieve different BCR- ABL levels, totally 16 sets and 24 samples per set were prepared. TRIzol reagent was added in each tube. Each laboratory tested BCR-ABL transcript levels of one set of samples. Agreement between BCR-ABLIS of each laboratory and PKUPH was assessed by the Bland- Altman method. For laboratories which did not meet the criteria of revalidation, linear regression equation was derived after the samples with maximum BCR-ABL deviation were removed until R²>0.98, then new CF was calculated.

RESULTS10 laboratories met the revalidation criteria with both bias within ±1.4 fold and 95% limits of agreement within ±6 folds, and their CFs still could be used for accurately conversion of BCR-ABLIS. New CFs were recalculated as of 1.8-6.3 folds of their previous CFs in 5 laboratories not met the criteria.

CONCLUSIONRevalidation of CF by sample exchange among laboratories was necessary for accurate and continuous application of BCR-ABLIS, which not only tested the validity of CF acquired before but also calculated new available CFs for those with invalid CFs.

Bone Marrow Cells ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; genetics

OBJECTIVETo investigate the roll of bone sialoprotein (BSP), a secreted glycoprotein, found in mineralized tissues in the development and progression of human esophageal squamous cell carcimoma (ESCC), and explore its association with clinicopathological characteristics and five-year survival of the patients.

METHODSThe expression of BSP was determined in 211 primary ESCC tumors and their paired nontumorous tissues using tissue-array, RT-PCR and immunohistochemistry.

RESULTSPrimary ESCC tissues showed a significantly higher expression rate of BSP mRNA than their paired nontumorous tissues (93.8% vs. 16.6%, P < 0.001), the same with BSP protein (56.9% vs. 31.3%, P < 0.001). The expression rate of BSP protein was correlated to lymph node metastasis and TNM stage (P < 0.05). The 5-year survival rate of BSP protein-positive ESCC patients was significantly lower than that of BSP protein-negative ESCC patients (P < 0.05). Multivariate analysis showed that tumor differentiation, TNM staging and BSP protein expression were independent factors affecting the prognosis of ESCC patients (P < 0.05).

CONCLUSIONSThe abnormal expression of BSP may play a significant role in the malignant progression and prognosis of ESCC, and BSP might be a marker reflecting the biologial behavior of ESCC.

Blotting, Western ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Esophageal Neoplasms ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Integrin-Binding Sialoprotein ; genetics ; metabolism ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; Survival Rate

OBJECTIVETo validate the conversion factor (CF) for the conversion of BCR-ABL (P210) transcript levels to the international scale in chronic myeloid leukemia (CML).

METHODSIn 2012, the international reference laboratory in Adelaide, Australia (IMVS) sent two batches of RNA samples, 30 samples per batch, to Peking University People's Hospital (PKUPH). By comparing BCRABL (P210) transcript levels reported by the two laboratories, CF of PKUPH was calculated and validated by IMVS. In 2013, PKUPH prepared the exchange samples for validation of CF of 9 hospitals who have calculated CFs before. The fresh BCR-ABL (P210) (+) cells were serially diluted by BCR-ABL (P210) (-) cells to prepare 22 kinds of samples with different BCR-ABL transcript levels, each kind had 10 parallel samples. Trizol reagent was added in each tube. Ten hospitals tested BCR-ABL transcript levels of one set of 22 samples. Agreement between BCR-ABL transcript levels of each laboratory and PKUPH was assessed by the Bland-Altman method.

RESULTSPKUPH successfully validated its CF with bias 1.1 fold and 95% limits of agreement between -4.7 and 4.9 fold. Of 9 hospitals whose validation performed by sample exchanges with PKUPH, 6 hospitals successfully validated their CF with bias ≤±1.4 fold and 95% limits of agreement within ±6 fold.

CONCLUSIONValidation of CF examined the stability of the detection of BCR-ABL (P210) transcript levels, which was necessary for the valid conversion of BCR-ABL (P210) transcript levels to the international scale in CML.

Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; standards ; Transcription, Genetic