Objective:To offset the shortage of traditional large-scale population surveillance and provide early-warning signals in the early stage of the outbreak of COVID-19, people in Chongqing had carried out severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) monitoring since 2023.Methods:After COVID-19 was managed with measures against Class B infectious diseases, we selected five sewage treatment plants with automatic sample collection facilities in four districts of the main city. Two samples of sewage from each sewage treatment plant were collected every week. Then SARS-CoV-2 from these samples was concentrated by aluminum hydroxide adsorption-precipitation, detected and analyzed by multiple real-time fluorescent RT-PCR.Results:From January 16 to December 31 of 2023, a total of 496 sewage samples were monitored, of which 285 samples were positive by SARS-CoV-2 nucleic acid assay, with a total detection rate of 57.46%. The detection rate of SARS-CoV-2 in weeks 3-5, 18-21 and 40-47 was 100.00%. The daily mean nucleic acid concentration of SARS-CoV-2 in sewage peaked in the 18th week, and then began to decline, entering a low level and fluctuated in epidemic period. The variable trend of daily mean concentration of SARS-CoV-2 nucleic acid was basically consistent with daily number of SARS-CoV-2 infected patients or SARS-CoV-2 positive rate in fever clinic counted by infectious disease monitoring system.Conclusions:The detection rate of SARS-CoV-2 in sewage of Chongqing is relatively high, especially in April to May, and sewage monitoring can indirectly reflect the status of COVID-19 infection.

Objective To investigate the molecular mechanism of translocation of heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1 )to the cytoplasm after glucose starvation and the effects of increased cytoplasmic translocation of hnRNPA2B1 on the survival of prostate cancer PC3 cells. Methods Human prostate cancer PC3 cells were divided into normal control group (cultured conventionally with glucose-containing medium,RPMI 1640 Medium)and glucose starvation group (cultured with glucose-free medium,RPMI 1640 Medium).The 2 types of cells were treated with deacetylase inhibitor,trichostatin A (TSA ) combined with nicotinamide (NAM),AKT inhibitor BEZ235,si-NC transfection,and si-hnRNPA2B1 transfection,respectively.Cytoplasmic and nuclear protein separation,immunoprecipitation and Western blotting were used to detect changes in hnRNPA2B1 acetylation,total AKT protein and its phosphorylation level,and expression levels of hnRNPA2B1 in the cytoplasm and nucleus.CCK-8 assay was employed to observe cell survival in each group.Results After 3～5 h of glucose starvation treatment,the acetylation of hnRNPA2B1 protein was reduced (P<0.01 ),and its cytoplasmic translocation was increased in PC3 cells (P<0.01 ),which was accompanied by enhanced AKT phosphorylation and activation of the AKT signaling pathway.TSA/NAM treatment,BEZ235 treatment,and si-hnRNPA2B1 transfection all resulted in obvious increase in acetylation of hnRNPA2B1 protein when compared with glucose starvation treated cells (P<0.01 ),which could inhibit the glucose starvation-mediated cytoplasmic translocation of hnRNPA2B1,suppress AKT phosphorylation,and consequently decrease the cell survival rate after glucose starvation (P<0.01).Conclusion Glucose starvation can maintain the survival of PC3 cells by inducing the activation of the Ac-hnRNPA2B1-AKT signaling pathway.

Objective:To investigate the optimal experimental conditions (including antigen retrieval time, antibody titers and antibody incubation time) for reliable detection of programmed death-ligand 1 (PD-L1) expression using PD-L1 (22C3) antibody concentrate, and to establish a laboratory developed test for PD-L1 detection.Methods:Using Dako PD-L1 IHC 22C3 pharmDX staining procedure and scoring guidelines as the standard reference (group A), the PD-L1 expression in 25 tissue specimens (including 15 lung cancer tissues, 5 tonsil tissues and 5 placenta tissues) was detected with Flex+/HRP detection kit (EnVision) under 8 different experimental conditions (groups B1 to B8). The staining results were then compared to those in group A.Results:In group B1, 3 tissue samples showed the percentages of PD-L1 positive tumor cells were similar to those in group A, while the percentages of PD-L1 positive tumor cells were lower than those in group A in the other samples. In group B7, two case showed a positive rate higher than that in group A that was also above the positive cut-off value, and the rest of the samples had a percentage of PD-L1 positive tumor cells slightly higher than that in group A, but still below the positive cut-off value. The staining results of group B8 were the closest to those of group A compared with the other groups. Although the percentages of PD-L1 positive tumor cells in the B2 to B6 groups were decreased in various degrees as compared with group A, they were still concordant with group A′s classification (positive vs. negative) and would not change the choice of clinical treatments.Conclusions:The experimental conditions are associated with detection rate of PD-L1 expression using 22C3 antibody. In the present study, the most-suitable alterative conditions in the PD-L1 detection using 22C3 antibody concentrate are those applied in the group B8 (including antigen retrieval in Dako PT Link tank at 97 ℃, pH 6.0 for 40 min and incubation with 22C3 antibodies (1∶100 dilution) at room temperature for 60 min, incubation with EnVision Flex+Linker at room temperature for 30 min, incubation with EnVision/HRP at room temperature for 30 min and DAB staining for 5 min), which could provide reliable results at minimum costs.

In the original publication the grant number is incorrectly published. The correct grant number should be read as "17140901600". The corrected contents are provided in this correction article. This work was partially supported by grants from the National Natural Science Foundation of China (Nos. 81670470 and 81600149), a grant from the Shanghai Municipal Commission for Science and Technology (17140901600, 18411953500 and 15JC1400201) and a grant from National Key Research and Development Program (2016YFC0905100).

Adenosine ; genetics ; Adenosine Deaminase ; genetics ; metabolism ; Animals ; CRISPR-Associated Protein 9 ; genetics ; metabolism ; CRISPR-Cas Systems ; genetics ; Embryo, Mammalian ; metabolism ; Gene Editing ; Genetic Variation ; genetics ; Mice ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of miR-132-3p on the proliferation of endothelial progenitor cells and its regulatory mechanism in order to provide a new theoretical basis for the treatment of deep venous thrombosis. Methods Real-time quantitative PCR (qPCR) was used to detect the expression of miR-132-3p in the plasma and endothelial progenitor cells of 27 healthy volunteers and 22 thrombus patients, and in endothelial progenitor cells under normoxic and hypoxic conditions. The miR-132-3p analogue and the specific siRNA were transferred into endothelial progenitor cells by the electroporation method. The effect of miR-132-3p on the proliferation of endothelial progenitor cells was detected using MMT and Cell Counting Kit-8 (CCK-8) methods. The effects of miR-132-3p on the expression of FOXO1 in endothelial cells were analyzed using the luciferase assay and western blots. Results The expression of miR-132-3p in clinical patients with thrombosis was significantly decreased to 0.45 ± 0.05 times of that of the healthy volunteers (P＜0.05). The expression of miR-132-3p in endothelial progenitor cells under hypoxia was down-regulated to (0.23 ± 0.13) times of that of under normoxia (P＜0.05). The expression of miR-132-3p of experiment group under hypoxia was up-regulated to (15.72 ± 2.06) times of that of control group (P＜0.05). MMT assay showed that the proliferation of cells in the experimental group under hypoxic condition was up-regulated to (7.79 ± 1.37) times of that in the control group (P＜0.01). CCK-8 assay showed that cell proliferation in experimental group was up-regulated to (6.46 ± 0.38) times of that in the control group (P＜0.01). Software analysis showed that FOXO1 was a direct target of miR-132-3p. Luciferase activity of miR-132-3p mimics transfected endothelial progenitor cells under hypoxic conditions were 0.47 times of that in siRNA treatment group. Western blot showed that the expression of FOXO1 protein in endothelial progenitor cells transfected with miR-132-3p mimics in hypoxia was 0.18 times of that in siRNA treatment group. Conclusions Compared with healthy volunteers, miR-132-3p expression in the blood of patients with thrombosis was significantly reduced that can promote transcription of the FOXO1 gene (and protein expression) and inhibit the proliferation of endothelial progenitor cells. It could be closely related to the formation of venous thrombosis.

Objective: To study the clinicopathologic characteristics, immunophenotype, pathologic diagnosis and differential diagnosis of myxoid adrenocortical adenomas. Methods: The clinical data, histological features and immunohistochemical results of 4 cases of myxoid adrenocortical adenomas were analyzed, which were collected from January 2014 to December 2016 at Guangdong General Hospital, with review of literature. Results: Four cases of myxoid adrenocortical adenomas were presented. The patients ages ranged from 26 to 45 years (mean =35 years). Microscopically, it showed a typical morphology, characterized by small-sized tumor cell cords or pseudo-glands embedded in an abundant extracellular myxoid matrix. Immunohistochemical staining showed tumor cells were strongly positive for Melan A, vimentin and focally for α-inhibin, one case showed strong and diffuse positivity for CAM5.2, and two cases showed diffuse positivity for synaptophysin, while negative for CgA, S-100 protein, epithelial antigen, CK7, CK20 and CKpan. Conclusions Myxoid adrenocortical adenomas are extremely rare, which may cause confusion with metastatic well-differentiated neuroendocrine tumours, sex cord-stromal tumoursor metanephric adenoma. Recognition of this entity would be beneficial for pathologists to avoid misdiagnosis, and unnecessary treatment.

Objective To investigate the effects of recombinant human growth hormone on prognosis,pulmonary function and immune function in the patients with pulmonary exogenous respiratory distress syndrome (ARDS).Methods Eighty-four cases of ARDS were selected and divided into the observation group (n=42) and control group (n=42) according to the random number table.The control group was treated with the routine therapy according to the ARDS Diagnosis and Treatment Guidelines,while on this basis the observation group used recombinant human growth hormone for conducting treatment.The treatment time lasted for 7 d.The clinical effects and improvement situation of immune function before and after treatment were recorded in the two groups.Results The Murray acute lung injury score,and acute physiology and chronic health status score (APACHE Ⅱ) after treatment in the observation group were lower than those in the control group (P<0.05).The mechanical ventilation time and ICU stay of observation group were shorter than those of the control group(P<0.05).The mortality rate of the observation group was lower than that of the control group(P<0.05).The levels of vital capacity (VC),total lung volume (TLC),forced expiratory vital capacity (FVC),forced expiratory volume of vital capacity (FEV1),forced expiratory vital capacity 1 second (FEV1/FVC) and carbon monoxide (DLCO) after treatment in the observation group were significantly higher than those in the control group (P<0.05).The levels of CD3+,CD4+ and CD4+/CD8+ after treatment in the observation group were higher than those in the control group (P<0.05).The levels of interleukin-6(IL-6),interleukin-8 (IL-8) and tumor necrosis factor-a (TNF-α) after treatment in the observation group were lower than those in the control group (P<0.05).Conclusion Recombinant human growth hormone can effectively improve the pulmonary function in the patients with pulmonary exogenous ARDS,improves the immune function and is conducive to the prognosis of patients.

Objective To systematically evaluate the influence of acupuncture-moxibustion on the peroneal nerve conduction velocity in type 2 diabetic peripheral neuropathy (DPN), and to provide clinical references for acupuncture-moxibustion treatment for DPN.Method By searching the CBM, CNKI, VIP, Wanfang, Pubmed, Springer and Medline databases, randomized controlled trials (RCT) of acupuncture-moxibustion for the type 2 DPN published from January 2000 to January 2014 were retrieved and the relevant data of the peroneal nerve conduction velocity were collected for methodological evaluation. RevMan 5.1 software was adopted to conduct the meta-analysis.Result Totally 10 RCTs were recruited with 685 cases involved, including 355 cases in the treatment group and 330 cases in the control group. The meta-analysis results indicated that acupuncture-moxibustion can produce a better effect in improving the motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) of peroneal nerve in type 2 DPN than the treatments used in the control group, and the differences were statistically significant between the two groups [MD=3.55, 95%CI (0.79, 6.31); MD=4.10, 95%CI (0.22, 7.99)].Conclusion Acupuncture-moxibustion can improve the peroneal nerve conduction velocity in type 2 DPN, and thus is worth application in clinic. Due to the limitation of the included studies, such as small sample size and low quality of the articles and high probability of bias, RCTs of large sample size and high quality are required to confirm the above conclusions.

Objective To analyze the resistant profile and genotypes ,as well as susceptibility to tigecycline in carbapenemase‐producing K lebsiella pneumoniae for better clinical management of such infections .Methods Nine strains of carbapenemase‐producing K .pneumoniae were isolated from clinical specimens during the period from February 2012 to May 2013 in our hospital .Bacterial identification and antimicrobial susceptibility testing were carried out with VITEK 2 Compact automatic microbiological assay systems .The phenotype of carbapenemase‐producing K . pneumoniae was detected by modified Hodges test .The genotype of carbapenemase was identified by PCR method .The susceptibility to tigecycline was tested by E‐test . Results All the 9 strains of carbapenemase‐producing K . pneumoniae were resistant to the 19 antibiotics tested .Modified Hodges test was positive for 7 strains (77 .8% ) .Target band of carbapenemase gene was identified in all the 9 strains of K . pneumoniae ,and all were confirmed as KPC‐2 gene .All the 9 strains were susceptible to tigecycline .Conclusions The resistance of carbapenemase‐producing K .pneumoniae is still serious .Tigecycline has shown good in vitro activity against such strains .