Objective To explore whether the effect of urine-derived stem cells(USCs)on chondro-cytes in osteoarthritis(OA)is related to the PI3K/AKT pathway.Methods Human urine was collected aseptically,and primary USCs were isolated and cultured through specific steps.Meanwhile,chondro-cytes were extracted from the knee joints of Sprague-Dawley rats to establish the OA model.USCs were co-cultured with OA chondrocytes by using Transwell co-culture system,and the transcriptome changes of chondrocytes before and after co-culture were analyzed by using the high-throughput RNA sequencing.Moreover,the phosphorylation levels of key proteins of the PI3K/AKT/mTOR pathway were detected using protein immunoblotting,and the PI3K agonist 740 Y-P was employed to further vali-date the relationship between the protective effect of USCs on OA chondrocytes,as well as its possi-ble relationship to the PI3K/AKT pathway and the possible mechanism.Results Transcriptomic analysis showed that the protective effect of USCs on OA chondrocytes might be related to the'PI3K-Akt sig-naling pathway'.Protein level analysis confirmed that USCs significantly inhibited the PI3K/AKT/mTOR pathway in OA chondrocytes.Moreover,protein immunoblotting and immunofluorescence showed that USCs improved the expression of OA-related proteins and autophagy-related proteins in OA chondro-cytes.What's more,the inhibitory effect of USCs on the PI3K/AKT/mTOR pathway was partially re-versed by using the PI3K agonist 740 Y-P,while the protective and autophagy-promoting effects of USCs on OA chondrocytes were down-regulated.Conclusion This study reveals the potential protective effect of USCs on OA chondrocytes through inhibition of the PI3K/AKT/mTOR signalling pathway and its close correlation with improved autophagy.Therefore,it provides a research basis and theoretical support for the future application of USCs in the treatment of OA.

Structural and functional explorations on bio-soft matter such as micelles,vesicles,nanoparticles,ag-gregates or polymers derived from traditional Chinese medicine(TCM)has emerged as a new topic in the field of TCM.The discovery of such cross-scaled bio-soft matter may provide a unique perspective for unraveling the new effective material basis of TCM as well as developing innovative medicine and biomaterials.Despite the rapid rise of TCM-derived bio-soft matter,their hierarchical structure and as-sembly mechanism must be unambiguously probed for a further in-depth understanding of their pharmacological activity.In this review,the current emerged TCM-derived bio-soft matter assembled from either small molecules or macromolecules is introduced,and particularly the unambiguous elucidation of their hierarchical structure and assembly mechanism with combined electron microscopic and spectroscopic techniques is depicted.The pros and cons of each technique are also discussed.The future challenges and perspective of TCM-derived bio-soft matter are outlined,particularly the requirement for their precise in situ structural determination is highlighted.

Objective To explore whether the effect of urine-derived stem cells(USCs)on chondro-cytes in osteoarthritis(OA)is related to the PI3K/AKT pathway.Methods Human urine was collected aseptically,and primary USCs were isolated and cultured through specific steps.Meanwhile,chondro-cytes were extracted from the knee joints of Sprague-Dawley rats to establish the OA model.USCs were co-cultured with OA chondrocytes by using Transwell co-culture system,and the transcriptome changes of chondrocytes before and after co-culture were analyzed by using the high-throughput RNA sequencing.Moreover,the phosphorylation levels of key proteins of the PI3K/AKT/mTOR pathway were detected using protein immunoblotting,and the PI3K agonist 740 Y-P was employed to further vali-date the relationship between the protective effect of USCs on OA chondrocytes,as well as its possi-ble relationship to the PI3K/AKT pathway and the possible mechanism.Results Transcriptomic analysis showed that the protective effect of USCs on OA chondrocytes might be related to the'PI3K-Akt sig-naling pathway'.Protein level analysis confirmed that USCs significantly inhibited the PI3K/AKT/mTOR pathway in OA chondrocytes.Moreover,protein immunoblotting and immunofluorescence showed that USCs improved the expression of OA-related proteins and autophagy-related proteins in OA chondro-cytes.What's more,the inhibitory effect of USCs on the PI3K/AKT/mTOR pathway was partially re-versed by using the PI3K agonist 740 Y-P,while the protective and autophagy-promoting effects of USCs on OA chondrocytes were down-regulated.Conclusion This study reveals the potential protective effect of USCs on OA chondrocytes through inhibition of the PI3K/AKT/mTOR signalling pathway and its close correlation with improved autophagy.Therefore,it provides a research basis and theoretical support for the future application of USCs in the treatment of OA.

Objective:To investigate the effect of ubiquitin binding enzyme 2T (UBE2T) on the radiosensitivity of lung adenocarcinoma and unravel its possible mechanism.Methods:A total of 45 patients pathologically diagnosed with different stages of lung adenocarcinoma and treated with radiotherapy in the Second Affiliated Hospital of Zunyi Medical University from March, 2019 to December, 2021 were enrolled, and the efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST1.1). All patients were divided into radiosensitive group ( n=25) and radioresistant group ( n=20). Radiosensitive group was complete remission (CR)+partial remission (PR), and radioresistant group was stable disease (SD) + progression disease (PD). Immunohistochemistry (IHC) was used to calculate the score based on the staining intensity and the number of positive cells. Chi-square test was combined to analyze the correlation between the expression level of UBE2T in paraffin specimens of lung adenocarcinoma patients and the radiosensitivity of patients. Lentivirus UBE2T-interfered (UBE2Tsh) A549 and UBE2T-overexpressed SPC-A-1 lung adenocarcinoma cells and their respective controls were constructed for irradiation and colony formation assay. The survivor fraction curve was fitted by single-hit multi-target model. The DNA double-strand break (DSB) marker γH2AX foci were detected by immunofluorescence (IF). The expression levels of UBE2T, γH 2AX and Rad51 proteins were detected by Western blot. Cell cycle and apoptosis rate of A549 were determined by flow cytometry. Binary variables were statistically analyzed by Fisher's exact probability method and measurement data were assessed by t-test. Results:High-expression level of UBE2T was correlated with the radiosensitivity of lung adenocarcinoma patients ( P<0.05). UBE2Tsh improved the radiosensitivity of A549 lung adenocarcinoma cells, and the sensitizing enhancement ratio (SER) was 1.795. UBE2T overexpression decreased the radiosensitivity of SPC-A-1 lung adenocarcinoma cells with an SER of 0.293. γH2AX foci number per cell were significantly increased in UBE2Tsh A549 cells after irradiation ( P<0.01) . Compared with the control group, the expression level of γH2AX protein was up-regulated ( P<0.01)and that of Rad51 protein was down-regulated in UBE2Tsh A549 cells after radiation ( P<0.001). Compared with the control group, the expression level of γH2AX protein was down-regulated ( P<0.05) and that of Rad51 protein was up-regulated in UBE2T overexpressed SPC-A-1 cells ( P<0.001). The proportion of UBE2Tsh A549 cells in G 2 phase was decreased ( P<0.01) and cell apoptosis was increased ( P<0.001). Conclusions:UBE2T might promote the radioresistance of lung adenocarcinoma cells by enhancing DNA DSB repair induced by radiotherapy, inducing cell cycle G 2 phase arrest, and reducing cell apoptosis.

Objective:To compare the efficacy, safety, and influencing factors among children with hepatitis B virus e antigen (HBeAg)-positive chronic hepatitis B (CHB) who received short-term therapy with pegylated interferon alfa-2a (Peg-IFNα-2a) or continuous therapy with entecavir (ETV).Methods:Quantitative data were compared using analysis of variance to compare the differences between groups. Enumeration data were compared by χ2 test (or Fisher's exact test). Univariate and multivariate logistic regressions were used to analyze the influencing factors. Results:Peg-IFNα-2a, ETV, and untreated group had HBsAg clearance rates of 46.2%, 5.3%, and 0 after 52 weeks of therapy, respectively. HBsAg clearance in the patients' group with Peg-IFNα-2a and ETV was all accompanied by anti-HBS positive conversion, and the difference was statistically significant ( χ2=13.616, P=0.001). Peg-IFNα-2a group was followed-up for 104 weeks. Peg-IFNα-2a, ETV, and the untreated group had HBsAg clearance rates of 46.2%, 10.5%, and 0%, respectively, and the differences were statistically significant ( χ2=11.056, P=0.004). Only one of the two children with HBsAg clearance in the ETV group had achieved anti-HBs antibodies, and the difference was statistically significant ( χ2=13.616, P=0.001). Univariate and multivariate logistic regression analysis showed that HBsAg clearance was associated with age and antiviral therapy. During treatment, adverse events such as fever ( n=4, 30.8%), rash ( n=4, 30.8%), fatigue ( n=1, 7.7%), leukopenia ( n=7, 53.8%), arthritis ( n=1, 7.7%), and alopecia ( n=3, 23.1%) were observed in the Peg-IFNα-2a group, while none were observed in the ETV group. Conclusion:Peg-IFNα-2a antiviral therapy produced higher HBsAg clearance than ETV in five-year-old and younger children with HBeAg-positive CHB, while ETV had fewer adverse events and was safer than Peg-IFNα-2a.

Objective:To analyze and summarize the characteristics of liver pathology and their relation to clinical markers and further explore noninvasive markers of liver fibrosis in children with chronic hepatitis B.Methods:Data of 80 hospitalized children with chronic hepatitis B who underwent liver biopsy without antiviral treatment from 2011 to 2020 were retrospectively analyzed. Inflammation and liver fibrosis characteristics were analyzed in children of different ages and genders. Variables with good correlation with liver fibrosis stage were selected to establish a non-invasive diagnostic score of liver fibrosis in children. Measurement data was used to compare the t-test or rank sum test. Mantel-Haenszel χ2 test was used for bidirectional ordered grouping data. Spearman’s rank correlation test was used for rank correlation analysis. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic value of the newly established diagnostic score in children with liver fibrosis. Results:The median age of the children was 6.4 years. HBV DNA level was high (P50 = 7.6 log 10 IU/ml), and serum alanine aminotransferase (ALT) in P50 was 171 U/L (< ULN: 5 cases, ULN-2ULN: 10 cases, > 2 ULN: 65 cases). Pathological analysis showed that the incidence of liver tissue inflammation was 97.5%, and the proportion of patients with G≥2 was 42.5%, while S≥2 was 36.3%. The incidence rate of liver fibrosis and liver cirrhosis was 81.3%, and 1.3%, respectively. The changes in liver tissue inflammation and fibrosis were gradually aggravated with the increase of age, and the proportion of high-grade inflammation and liver fibrosis in male children was higher than that in female children. Serum levels of glutamyl transpeptidase (GGT), γ-glutamyltransferase/platelet ratio (GPR) and HBeAg had a good correlation with fibrosis stage ( rs = 0.397, 0.389, and - 0.311) in children with chronic hepatitis B. The combination of GGT, GPR and HBeAg can establish a non-invasive diagnostic score for evaluating liver fibrosis in children. When the score is less than 1.5, it can be diagnosed as S0, and 1.5 ≤ score < 3.5, it can be diagnosed as S1; 3.5 ≤ score < 5.5, the diagnosis of fibrosis is S2; score≥ 5.5, the diagnosis of fibrosis is S≥3. The sensitivity and specificity were 80%, 83%, 86%, and 53%, 55%, 67%, respectively. Conclusion:The incidence of liver tissue inflammation in children with chronic hepatitis B with elevated and fluctuating transaminase levels is high, and the pathological changes of liver tissue aggravate with the age of the children. GGT, GPR and HBeAg have a good correlation with liver fibrosis in children with chronic hepatitis B. Therefore, combining the above-mentioned markers to establish a new noninvasive diagnostic score has certain diagnostic value for liver fibrosis stage S0-S3 in children with chronic hepatitis B.

Objective:To compare the baseline difference in the quantitative hepatitis B core antibody levels (qAnti-HBc) between non-response and response group in children with HBeAg-positive chronic hepatitis B (CHB) who received antiviral therapy, and further explore the proportion and functional activity of CD8 + memory T lymphocyte subsets with different qAnti-HBC levels in peripheral blood of children.Methods:The baseline anti-HBc quantification (qAnti-HBc) levels of 85 children with HBeAg-positive CHB who visited the Department of Infectious Diseases, Children's Hospital of Chongqing Medical University from June 2018 to December 2020 were detected retrospectively. The relationship between the baseline qAnti-HBc level and HBeAg serological response in 37 children who received antiviral therapy was analyzed. The proportion of CD8 + memory T lymphocyte subsets and the secretion levels of interferon (IFN) γ, and tumor necrosis factor (TNF) α in peripheral blood of 59 children at baseline were detected by flow cytometry. The relationship between qAnti-HBc level and the proportion and functional activity of CD8 + memory T lymphocyte subsets was analyzed. Pearson’s Chi-square test was used to compare the count data. Mann-Whitney U test or Kruskal-Wallis test was used to compare measurement data between two or more groups, and Spearman’s rank correlation analysis was used for the correlation between continuous variables. Results:Among 37 children who received entecavir (ETV, 21/37 cases) or pegylated interferon (Peg-IFN, 16/37 cases), 18 cases had developed HBeAg seroconversion (10/ 21 cases in the ETV group, 8/16 cases in the Peg-IFN group). The baseline qAnti-HBc level was significantly higher in the response group [4.71 (4.64~4.81) log 10IU/ml] than the non-response group children [4.54 (4.45~4.64) log 10IU/ml, Z = -3.316, P = 0.001]. The proportion of CD8 + Tem, CD38 +CD8 + Tem, CD38 +CD8 + Temra cells and the levels of IFNγ and TNFα secreted by CD8 + T lymphocytes were significantly higher in the high-qAnti-HBc group than the low-qAnti-HBc group ( P < 0.05). The proportion of CD8 + Tem, CD38 +CD8 + Tem and CD38 +CD8 + Temra cells was significantly higher in ALT > 1× upper limit of normal value (ULN) group than ALT≤1×ULN group ( P < 0.05). However, there were no significant differences in the levels of IFNγ and TNFα secreted by CD8 + T lymphocytes between the two groups ( P > 0.05). Spearman’s correlation analysis showed that qAnti-HBc was positively correlated with the proportion of CD8 + Tem, CD38 +CD8 + Tem, CD38 +CD8 + Temra cells and the level of IFNγ secreted by CD8 +T lymphocytes ( P < 0.05). Additionally, ALT was only positively correlated with the proportion of CD38 +CD8 + TEM and CD38 + CD8 + Temra cells ( P < 0.05). Conclusion:Raised baseline qAnti-HBc level is related to the HBeAg serological response to antiviral therapy in children with CHB. Peripheral blood effector CD8+ T lymphocytes of CHB children with higher qAnti-HBc show stronger phenotype and functional activation characteristics, which may shed some light on the underlying immune mechanism related to antiviral therapy efficacy in children with CHB.

A patient with thymoma associated immunodeficiency syndrome (Good's syndrome) and bronchiectasis was retrospectively analyzed. Good's syndrome is a rare condition of immunodeficiency that is characterized by thymoma and hypogammaglobulinemia. It is important to bear in mind that Good's syndrome should be included in the differential diagnosis When patients repeatedly visited for bronchiectasis or infection, we should alert to their immune state and history of thymoma. Early screening of immunological status and aggressive correction of immune deficiency are beneficial to improving the prognosis to patients with Good's syndrome.
Agammaglobulinemia/complications* ; Bronchiectasis/complications* ; Humans ; Retrospective Studies ; Thymoma/complications* ; Thymus Neoplasms/complications*

Objective: To understand the attitude of college students in Zhejiang towards e cigarette and its influencing factors, so as to provide basis for making prevention and control strategies. Methods: In September 2020, 10 colleges and universities in Zhejiang Province were selected to conduct an online survey by a combination of typical sampling method and convenience sampling method. A multivariable Logistic regression model was used to analyze the influencing factors. Results: Among 884 subjects, 93 (10.52%) were positive about e-cigarettes, 310 (35.07%) thought e-cigarettes were harmless, 252 (28.51%) thought e-cigarettes were not addictive and 67 (7.58%) of respondents were using e-cigarettes. Multivariate Logistic regression analysis showed that gender, grade, cost of living, whether or not smoking e-cigarettes were harmful to college students attitudes towards e-cigarettes( OR =0.59, 0.47, 1.87, 0.34, 0.54, P <0.05). Conclusion College students in Zhejiang Province have a positive attitude towards electronics and are not active in avoiding the dangers of smoking. To make full use of the work of concentrated trainees in tobacco control, efforts should be made to break the positive image of e-cigarettes. Junior college students should be the prioritized population for intervention, and female students should not be neglected.

Objective To analyze the drug resistance phenotype and genetic background of New Delhi metallo-β-lactamase (NDM)-producing bacterial strains in Lishui area of Zhejiang province.Methods The imipenem-resistant Enterobacteriaceae strains were isolated from January 2012 to December 2016 in Lishui Municipal Central Hospital of Zhejiang Province.Mrieux Vitek 2 Compact system was used to identified strains and PCR was used to screen for blaNDMgene.Susceptibility was detected by K-B method and MICs were obtained by Vitek 2 with GN13 cards.Plasmids typing was carried out by DNA sequencing of the replication initiator with the transconjugates as templates.The blaNDMgenetic contexts were detected by PCR and complete genome sequencing.Results A total of 102 strains of carbapenem-resistant enterobacteria (CRE), mainly Escherichia coli and Enterobacter cloacae, were isolated, of which 15 were positive for blaNDMwith a positive detection rate of 14.7%.The resistance rate to β-lactam antibiotics was 100%, and the resistance rates to aztreonam, compound sulfamethoxazole , tobramycin and gentamicin were all ＞80%;and the resistance rate to quinolones was ＞50%.Among the 15 NDM-producing strains , 12 strains were positive for Hodge test, and 2 strains of Enterobacter cloacae and 1 strain of Escherichia coli were negative. There were 11 strains of blaNDM-1and 4 strains of blaNDM-1.A variety of plasmid types such as IncX 3, IncFIIγ and IncA/C were detected, and 4 blaNDM-5genes were located in the IncX3 plasmid.Four blaNDMsurrounding gene structures were found, of which 8 blaNDM-1genes were located in ISAb125-hyp-blaNDM-1-bleMBL-TrpF-DsbC-IS26, while blaNDM-5was located in IS3000-IS5-blaNDM-5-bleMBL-TrpF-DsbC-IS26 structure, and the former was reported for the first time in China.Conclusion NDM-producing bacterial strains in Lishui area are prevalent at a low level and have high sensitivity to fosfomycin and polymyxin .The blaNDMgene may have multiple sources, but IncX3 plasmid is still the main source for gene transfer.Some new types of blaNDM-1 gene structure have been also found in this study.