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 evaluate the application of abbreviated protocol (AP) of magnetic resonance imaging (MRI) in screening of breast cancer with dense breast tissue.Methods Total 478 patients with dense breast tissue,who had negative X-ray mammography (MG),underwent dynamic contrast-enhanced MRI (DCE-MRI).The AP MRI,which consisted of first postcontrast subtracted (FAST) and maximum-intensity projection (MIP),and full diagnostic protocol (FDP) MRI images were analyzed by Breast Imaging Reporting and Data System (BI-RADS).Among 478 patients the histopathological diagnosis was available in 39 cases (41 breast lesions).With pathological diagnosis as gold standard,the sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV) of AP and FDP in breast cancer detection were evaluated.Results In 41 breast lesions there were 16 malignant lesions and 25 benign lesions.FDP detected all 16 malignant lesions,including 9 cases of ductal carcinoma in situ,6 cases of invasive ductal carcinoma and 1 case of mucinous carcinoma;AP detected 15 out of 16 cases of breast cancer.The sensitivity of FDP and AP in detection of breast malignant lesions was 16/16 and 15/16,respectively (x2 =0.725,P =0.224).The specificity of FDP and AP was 92％ (23/25) and 76％ (19/25),respectively (x2 =6.327,P =0.012).Conclusion For women with dense breast tissue AP of MRI can be used in early screening of breast cancer.

SUMMARY Thehumanembryonicstemcells(hESCs)serveasaself-renewable,genetically-healthy, pluripotent and single source of all body cells,tissues and organs.Therefore,it is considered as the good standard for all human stem cells by US,Europe and international authorities.In this study,the standard and healthy human mesenchymal progenitors,ligament tissues,cardiomyocytes,keratinocytes,primary neurons,fibroblasts,and salivary serous cells were differentiated from hESCs.The human cellular health-safety of NaF,retinoic acid,5-fluorouracil,dexamethasone,penicillin G,adriamycin,lead ace-tate PbAc,bisphenol A-biglycidyl methacrylate (Bis-GMA)were evaluated selectively on the standar-dized platforms of hESCs,hESCs-derived cardiomyocytes,keratinocytes,primary neurons,and fibro-blasts.The evaluations were compared with those on the currently most adopted cellular platforms.Parti-cularly,the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endo-thelial cells (HUVECs)were evaluated.The results showed that the standardized hESCs cellular plat-forms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Objective To discuss the treatment method and opportunity for patients with gallbladder stones and extrahepatic bile duct stones who failed endoscopic removal of common bile duct stones by endoscopic retrograde cholangiopancreaticography (ERCP). Methods 12 patients, with gallbladder stones and extrahepatic bile duct stones, failed endoscopic stone extraction (ESE), underwent emergency one-stage laparoscopic cholecystectomy (LC) and Laparoscopic common bile duct exploration (LCBDE). Results All of the patients were successfully completed LC +LCBDE and stones were completely removed. Hyperamylasemia occurred in 3 cases and there was no bile leakage, intestinal leakage, cholangitis, pancreatitis, biliary bleeding and other complications. Conclusions Emergency LCBDE has been shown to be a safe and effective salvage procedure for failed ESE.

Objective To explore the multi-modal MRI characteristics of breast cancers in dense breasts.Methods 120 patients with breast cancer shown on mammography underwent breast MRI,which were solitary and confirmed by pathological examination. According to the BI-RADS classification of breast,the 120 cases were divided into two groups including dense type breast and non-dense type one.The differences in morphological features,ADC values (b=1 000 s/mm2 )and time-signal intensity curve (TIC)of the lesions between two groups were analyzed and compared.Statistical analysis was performed using SPSS1 6.0.Results The breast cancers in dense breast were vulnerable to have a spiculated margin (44/68 in the dense breast group vs.1 6/52 in the non-dense breast group,P =0.000).The size of the lesion in dense breast (1.83 ±0.98)cm was bigger than that in non-dense breast (1.40±0.46)cm (P =0.005).The non-mass-like enhancement of the lesion in dense breast was much more than that in non-dense breast (P =0.000).In addition,the average ADC values of the lesion in dense breast (0.89±0.12)×10 -3 mm2/s were lower than in non-dense breast (0.95±0.10)×10 -3 mm2/s(P =0.01 6).Conclusion The breast cancer in the dense breast has different MRI findings in comparison with non-dense breast.

Objective To explore the method for early diagnosis of masked polycythemia vera (mPV) by comparing and analyzing the features of mPV and polycythemia vera (PV). Methods A total of 200 newly diagnosed male patients were collected based on diagnostic criteria, including 100 cases of PV and 100 cases of mPV. Erythropoietin (EPO), neutrophil alkaline phosphatase (NAP) score, bone marrow biopsy (BMB), and JAK2 V617F mutation were evaluated in all cases. After 6 months, hemoglobin (Hb) and JAK2 V617F mutation load were detected in patients without special treatment in two groups during follow-up. Results EPO, NAP score, BMB hematopoietic volume and the number of megakaryocyte had no statistical differences between PV group and mPV group [(3.4 ± 0.7) U/ml vs. (3.2±0.6) U/ml, (276±20) score vs. (278±21) score, (78±10) % vs. (76±9) %, (53±6) vs. (51±5), respectively], while JAK2 V617F mutation load in PV group was higher than that in mPV group[(89.2±9.4) % vs. (78.1±8.6) %, P<0.05]. In mPV patients without special treatment, Hb ≥185 g/L was found in 37 patients after 6 months, and the level of Hb and JAK2 V617F mutation load in these 37 patients reached (194±8) g/L and (90.7±9.1) %, respectively. Conclusions There is no significant difference in EPO, NAP score and BMB between PV and mPV, but the JAK2 V617F mutation load is different. In mPV patients without special treatment, Hb level can reach the typical PV diagnostic criteria after 6 months, meanwhile, JAK2 V617F mutation load is also increased.

Objective To evaluate the value of proton magnetic resonance spectrum (1H-MRS) in the differential diagnosis of Alzheimer's disease (AD) and vascular dementia (VD) patients with white matter lesions (WMLs).Methods Multi voxel 1H-MRS were performed to 30 patients with AD,30 patients with VD,and 30 normal control volunteers.The levels of the N-acetylaspartate (NAA),myo-inositol (mI),glutamate (Glu),and creatine (Cr) were measured in the white matter adjacent to the lateral ventricles.The ratios of NAA/Cr,mI/Cr and Glu/Cr were calculated and compared among the group.Comparisons of the 1H-MRS indexes among AD,VD and NC group were conducted by one-way ANOVA.Results NAA/Cr ratios of the region of interesting in AD (1.59±0.15)and VD(1.53±0.12) group were significantly decreased compared with NC (1.79±0.22)group (P＜0.05),and Glu/Cr ratios of the region of interesting in AD(0.41±0.03) and VD(0.49±0.04) group were significantly decreased compared with NC(0.57±0.05) group (P＜0.05).However,there were not significant differences for NAA/Cr ratios of the paraventricular white matter region between AD and VD group (P＞0.05),and Glu/Cr ratios in AD and VD groups were significantly different (P＜0.05).A significantly increased mI/Cr was found in AD (0.68±0.05) group than in VD(0.57±0.04) and NC(0.55±0.03) groups (P＜0.05),and there were no significant changes in mI/Cr ratios between VD and NC groups (P＞0.05).Conciusion Multi-voxel 1H-MRS is an effective method in the differential diagnosis of dementia,and changes of the mI/Cr and Glu/Cr ratios of the white matter adjacent to the lateral ventricles may help to identify the AD and VD patients.

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on type 2 diabetic mice model and to provide mechanistic insights into its therapeutic effect. Type 2 diabetic animal model was established with high calorie fat diet and low dose streptozotocin (STZ) injection. Mice were then randomized into 5 groups: model control, FGF21 0.25 and 0.05 μmol x kg(-1) x d(-1) groups, insulin treatment group. Ten age-matched normal KM mouse administered with saline were used as normal controls. Serum glucose, insulin, lipid products and the change of serum and liver tissue inflammation factor levels between five groups of mouse were determined. The results showed that blood glucose, insulin, free fatty acids (FFAs), triglycerides, and inflammatory factor average FGF-21 of type 2 diabetes model group and normal control group were significantly higher (P < 0.01), while compared with insulin group, no difference was significant. Average blood glucose, insulin, blood lipid and inflammatory factor of FGF-21 treatment group compared with type 2 diabetes group was significantly lower (P < 0.01) and insulin group has no difference with the model control group. The results of OGTT and HOMA-IR showed that insulin resistance state was significantly relieved in a dose-dependent manner. Thus, this study demonstrates that FGF-21 significantly remits type 2 diabetic mice model's insulin resistance state and participates in the regulation of inflammatory factor levels and type 2 diabetes metabolic disorders.

Objective To construct recombinant MVA vaccine encoding codon-optimized HIV-1 CRF01_AE gp145 gene and evaluate its immunogenicity in mice.Methods The intracellular region of codon-modified HIV-1 CRF01_AE consensus env gene obtained in our preliminary study was removed to get gp145 gene.The modified gp145 gene was cloned into shuttle plasmid pSC11.BHK-21 cells infected with wild-type MVA in advance were transfected with the recombinant shuttle plasmid.Then the recombinant MVA vaccine expressing gp145 rMVA-AEgp145 was obtained by homologous recombination.After expression of Gp145 was confirmed by WB assay,BALB/c mice were immunized with the recombinant MVA vaccine and the cellular immune responses were evaluated.Results Recombinant MVA vaccine expressed Gp145 protein efficiently and induced high level of cellular immune responses in immunized BALB/c mice.Conclusion The recombinant MVA vaccine encoding HIV-1 CRF01_AE gp145 gene was constructed successfully.This vaccine could induce strong env-specific CTL responses in mice.

Objective To evaluate target gene IL15 expression by lentiviral vector in mammalian cells,we constructed a recombinant lentivirus expressing human interleukin 15 to be used as vaccine adjuvant or in gene immunotherapy.Methods The IL15 gene was cloned from human peripheral blood mononuclear cell (PBMC) and constructed into plasmid plvx-IL15-Ires-gfp.Recombinant lentivirus was produced in 293T cells and immature dendritic cells were infected in vitro.Results The target gene expression was detected,cell lines stably expressing IL15 were obtained by antibiotic screening.Immature dendritic cells can also be infected and express IL15.Conclusion We successfully constructed lentivirus vector that was able to express ILl5 in dendritic cells without inducing maturation,it could be used in gene immunotherapy or as a vaccine adjuvant.