ObjectiveThis study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial. MethodFrom October 13^th， 2021 to December 25^th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score. Result（1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）. ConclusionTibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.

Objective:To investigate the changes of regulatory B cells (Bregs) and lymphocyte subsets in children with primary immune thrombocytopenia (ITP), and to analyze their relationship with ITP duration and remission time.Methods:Proportion of different immune cell subsets were detected in the peripheral blood of 88 ITP children before treatmen, 84 ITP children after treatmen and 45 normal controls by flow cytometry. The treatments included glucocorticoids, intravenous IgG as first-line treatment. The changes of lymphocyte subsets in ITP children with different remission time after treatment were compared.Results:The Th, natural killer (NK), Breg cell counts and Th/Tc ratio in all children with ITP were significantly lower than those in the control group ( P<0.05), while the Tc and CD19 + B cells were significantly higher than those in the control group ( P<0.05). The Breg cell count in the persistent and chronic ITP groups was significantly lower than that in the newly diagnosed ITP ( P<0.05). A total of 84 children with ITP were relieved after treatment: 58 cases (69.1%) achieved long-term remission, and 26 cases (30.9%) had short-term remission. The Breg and lymphocyte subsets in the short-term remission ITP group after treatment were not significantly different from those before treatment, but there were still significant differences from the control group ( P<0.05); The Breg and lymphocyte subpopulations in the long-term remission ITP group after treatment were not significantly different from those in the control group, but compared with those before treatment, other cell subpopulations except Tc were significantly increased ( P<0.05); The NK cells and Breg cells in the long-term remission ITP group were significantly higher than those in the short-term remission ITP group ( P<0.05). Conclusions:The Bregs in peripheral blood of long-duration ITP children significantly decrease. After treatment, the Bregs in ITP children with long-term remission increase significantly, indicating that the restoration of the immune mechanism of Bregs may be related to the long-term remission of children with ITP.

The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²⁺ or 0.1 mmol/L Cu²⁺ treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.
Animals ; Cyclization ; Endo-1,4-beta Xylanases ; chemistry ; metabolism ; Enzyme Stability ; Industrial Microbiology ; methods ; Microbiota ; Protein Engineering ; Rumen ; enzymology ; microbiology ; Temperature

Objective:To investigate the anti-leukemia effect of icaritin on 697 cells of children relapsed acute lymphoblastic leukemia in vitro, and to analyze its possible mechanism. Methods:697 cells were cultured in vitro and divided into control group and icariin groups (5, 10, 20 μmol/L). After treated with icaritin, the cell proliferation was detected with cell counting kit-8 (CCK-8) method, and apoptosis was measured with Hoechst 33258 staining and Flow Cytometry Assay. Expression of Bcl-2, Cyt-C, Bax and Caspase3 proteins in 697 cell were detected by Western blot. Results:After treatment with various concentrations (5-20 μmol/L) of icaritin for different time-points (24, 48, 72 h), the growth inhibition rate exhibited a dose and time-dependent manner ( r=0.76, r=0.89); the apoptotic rate exhibited significant increase for 48 h ( P<0.05). Bcl-2 protein decreased significantly ( P<0.05), while Bax, Cyt-C and Caspase3 cleavage protein increased significantly in icaritin treated group for 48 h compared with control group ( P<0.05). Conclusions:Icaritin shows the effects of anti-leukemia by inducing apoptosis and inhibiting proliferation of 697 cell. Icaritin maybe induce apoptosis of 697 cell by down-regulation of Bcl-2 expression and up-regulation of the Bax, Cyt-C and cleaved Caspase 3 expression.

Objective To analyze MRI findings and pathological data of intracranial solitary fibroma tumor(SFT)to improve the understanding of this disease and the level of imaging diagnosis.Methods MRI images and clinical pathological data of 6 patients with intracranial SFT confirmed by surgery and pathology were analyzed retrospectively.Results All tumors were located in the supratentorium,in which 4 cases were irregularly lobulated,2 cases were oval.All lesions were clear boundaries,and size 3.0-8.6 cm (an average of 6.3 cm).The tumor was located in the right fronto-perietral area in 1 case,cross of the frontal midline in 2 cases,cross of the occipital midline in 2 cases and cross of the cerebellum in 1 case.On T1WI,6 cases of tumor parenchyma showed slight low signal.On T2WI,4 cases of tumors showed mixed signal with the"Yin and Yang"sign,and peritumoral edema was showed in 4 cases, tortuous flow void blood vessels were showed in 5 cases.On FLAIR,mixed signal was showed.On DWI,slightly higher signal was showed.On MRI enhanced study,tumor parenchyma was significantly increased,in which 3 cases of T2WI low signal areas were significantly enhanced,cystic necrosis and edema were not enhanced,and 3 cases showed"dural tail sign".On pathological study, tumor cells were spindle-shaped,bundle and braided distribution,rich in collagen fibers.Immunohistochemistry showed CD34(+), CD99(+),Vim(+).1 case was malignant,and 1 case was potentially malignant.Conclusion MRI features of intracranial SFT have certain characteristics,it is difficult to be characterized qualitatively.MRI has obvious advantages in the relationship of tumor location, shape,range and its relationship with surrounding tissue.It has important clinical application value for the clinical operation plan and the residual or recurrence of tumor after operation.

Objective To investigate the changes of T cells and dendritic cells (DCs) and their related factors in children with immune thrombocytopenia (ITP) before and after therapy,and to analyze their clinical significance.Methods T-cells and DCs subsets were determined by flow cytometry both in 64 children with ITP (ITP group) before and after therapy and the control group.The serum levels of interleukin (IL)-4,interferon-γ (IFN-γ),transforming growth factor beta 1 (TGF-β1),and IL-27 were detected by enzyme linked immunosorbent assay (ELISA).Results Treatments of glucocorticoid or IVIg were effective in 41 cases of 64 ITP children.Compared to the control group,helper T cells (Th),Th/suppressor T cells (Ts),T regulatory cells (Treg),plasmacytoid DC (pDC),pDC/myeloid DC (mDC),and TGF-β1 in ITP patient group before treatment were significantly lower,while IFN-γ and IFN-γ/IL-4 were significantly higher (P ＜0.05).In ITP group,Th,Th/Ts,Treg,pDC,pDC/mDC,TGF-β1,and IL-27 were significantly increased,while IFN-γ and IFN-γ/IL-4 were decreased in children with ITP after therapy and achieved response (P ＜ 0.05).However,there was no significant difference between before and after therapy in ITP children without treatment response (P ＞ 0.05).Conclusions T cells and DCs subsets disorder and abnormal cytokine levels are observed in children with ITP,which can be corrected by immunosuppressive therapy,indicating that Th1 overactivity and the decrease of Treg and pDC both in quantity and function may be related to the pathogenesis of ITP in children.

Objective To explore the safety of the human bone marrow‐derived mesenchymal stem cells (hBMSCs) after silencing of human leukocyte antigen A2 expression .Methods We divided the cells into three groups:normal cultured cells of the 8th passage served as control group , and hBMSCs after HLA A2 silencing expression of the 5th and 15th passage as experimental groups 1 and 2 ,respectively .The hBMSCs were recultured by sterile methods .The growth curve ,telomerase activation ,and expressions of P27 ,cyclin D2 and cyclin‐dependent kinase 4 (CDK4) were utilized to explore the safety of the hBMSCs induced by LV‐siRNA‐HLA A2 .The BMSCs were transplanted to the subcutaneous layer of nude mice .Tissue types were detected 24 weeks after transplantation . Results The cell curves had no obvious left or right shift in all the groups . The telomerease activation in experimental groups 1 and 2 did not significantly differ from those in control group . The expressions of anti‐oncogene P27 ,cyclin D2 and CDK4 had no obvious difference between the two experimental groups and control group , either . There was only ectopic osteogenesis 24 weeks after the BMSCs (HLA A2 gene silenced ) were transplanted to the subcutaneous layer of the nude mice .Conclusion There was no obvious evidence to support that hBMSCs had undergone change in safety after the silencing of HLA A 2 expression .

Objective To investigate the impact of negative perfectionism on negative emotion(anxiety,depression) and the moderating effects of positive perfectionism.Methods A study was designed and a sample of 380 college students completed questionnaires including PANPS,SAS and SDS.Results ① Correlation analysis indicated positive perfectionism was negatively correlated with anxiety and depression (r1 =-0.25,P1 ＜0.01,r2 =-0.29,P2＜0.01),while negative perfectionism was positively correlated with anxiety and depression(r1 =0.26,P1 ＜0.01,r2 =0.22,P2＜0.01).② Moderating effects analysis indicated that positive perfectionism significantly moderated the regulating effect of negative perfectionism upon negative emotions(anxiety,depression) (β 1 =-2.64,β2 =-7.67,P＜0.01).Conclusion These findings suggest that the higher level of positive perfectionism,the greater influence of perfectionism on depression and anxiety,and positive perfectionism could buffer the negative perfectionism on anxiety and depression.

Objective To measure the costs of primary health services in the community for developing government compensation standard and prediction of budget in 201 5.Methods Literature review and panel discussions were used to build the index database.A two-round Delphi expert consultation determined the work to do and steps for community healthcare standards.60 community health centers were sampled by stratified random sampling for survey,and the work hours,workload and service volume of respective services in 2013 were measured according to the service standards of primary care.Results The community primary care consists of the categories of outpatient services,nursing care, laboratory tests and drug management,totaling 20 services and 88 working procedures.The total work hours of primary healthcare service at 60 community health centers were 2 557 187.9 hours,which is adjusted to 2 959 21 5.1 hours based on workload coefficients.Based on the income standards of employees at the community health centers investigated,and the human cost price which was recommended by the experts,the total costs of primary healthcare of the 60 centers were 245 million,248 million and 318 million respectively,and the average cost was 71.0 yuan,71.8 yuan and 92.1 yuan per visit respectively.By such standards,the government should subsidize 38.9 ~ 60.0 yuan per visit to the centers.The total expenditure of primary healthcare of all communities in Shenzhen in 201 5 was predicted to be 314 to 407 million,for which the government is expected to subsidize 1.68~2.58 billion.Conclusion The subsidies for community primary healthcare fall short as the cost per visit runs up in 2013 to 71.0 yuan per visit or more,far above the current subsidy of 32.1 yuan per person.The subsidy per visit should be made 50.0 yuan in 201 5.

Objective The study was designed to investigate the serum levels of HE4 in patients with lung cancer,and explore its prognostic value.Methods Blood samples of 106 healthy adults and 191 patients with lung cancer before treatment were measured by means of ELISA for HE4 levels in different stages and pathological types,for analyzing prognostic effect of HE4.Results The serum levels of HE4 in patients with lung cancer (median level 91.63 pmol/L) were significantly higher than control group (median level 56.42 pmol/L) (U =3 081.00,P ＜ 0.05).AUC of serum HE4 was 0.85,with a cut-off value of 82.70 pmol/L (specificity =95.31％,sensitivity =62.32％).HE4 expression was not statistically related to clinical stage and pathological types of lung cancer.The median overall survival (mOS) was 36.87 months in the HE4-low group and 30.43 months in the HE4-high group (P ＜0.05),respectively.HE4 level was an independent prognostic factor for overall survival (HR =2.15,95％ CI 1.49-3.12,P ＜ 0.05).Conclusions The prognosis of patients with high HE4 expression is poor.Therefore,it might be necessary for patients with high level of HE4 to receive more aggressive adjuvant therapy.