OBJECTIVE:To determine residual citrinin in Zhibituo tablet by solid phase extraction-UPLC-MS/MS. METH-ODS:The sample was processed preliminary by solid extraction. The determination was performed on Zorbax Eclipse Plus C18 col-umn with mobile phase consisting of water (0.1% formic acid)-acetonitrile (70:30,V/V,gradient elution) at the follow rate of 0.30 mL/min. The column temperature was 40 ℃,and analysis time was 7 min,the sample size was 2 μL. Ionization mode was positive ion mode. The ion source temperature was 150 ℃,drying gas velocity was 15 L/min,sheath gas temperature was 350 ℃, sheath gas flow rate was 12 L/min,capillary voltage was 3500 V,atomization device pressure was 40 psi,atomization voltage was 0 V. The acquisition mode was MRM mode. RESULTS:The linear range of citrinin was 0.1-20 ng/mL(r=0.9994);the limits of quantitation and detection were 0.05 and 0.01 ng/mL. RSDs of precision,stability and repeatability tests were all lower than 2.0%;recoveries were 98.868%-103.160%(RSD=1.5%,n=6). CONCLUSIONS:The method is rapid,sensitive,accurate and suitable for the determination of residual citrinin in Zhibituo tablet.

Wnt signaling pathway has a critical role in differentiation, proliferation, supersession and metabolism of osteocyte. Although the principal components are relatively simple, however, the numerous components consisted of Wnt signaling participate in the reaction, including several LRPs ( low density lipoprotein receptor-related protein,LRP) , 19 different Wnt proteins and 19 different FZDs( frizzled receptor, FZD) , and non-canonical Wnt signaling and multiple inhibitors of Wnt signaling.The Wnts linked to mem-brane coreceptor and activated downstream of canonical Wnt signaling or non-canonical Wnt signaling, In addition, different Wnts had different roles in differentiation, bone formation and bone absorption of osteocyte.This review aims to understand the relationship be-tween Wnt signaling and osteocyte metabolism and bone diseases, by analyzing a variety of factors and the process of Wnt protein and Wnt signaling pathway in activity of osteocyte.

Objective:To investigate the feasibility of real-time fluorescence quantitative PCR in the identification of Fritillariae Cirrhosae Bulbus.Methods:The DNA of samples was extracted by magnetic beads ,the primers were amplified by real-time fluores-cence quantitative PCR , and the Cq value and amplification curve were used to determine the samples .Results:The Cq values for five batches of Fritillariae Cirrhosae Bulbus were lower than 30, and the curve had obvious growth period .No Cq value was shown for Fritil-lariae Ussuriensis Bulbus , Fritillariae Thunbergii Bulbus and Bolbostemmatis Rhizoma with straight line curves .Conclusion:The meth-od is simple,feasible and effective in the identification of Bulbus Fritillariae Cirrhosae with high accuracy and good reproducibility .

Objective With the development of minimally invasion and the popularization of arthroscopy the technology of ar-throscopy has been widely applied for the treatment of acute and the safety of this has been validated.To analyze the clinical effect of ar-throscopically assisted double Endobutton plate reconstruction of coracoclavicular ligament in the treatment of acute acromioclavicular joint dislocation. Methods From November 2013 to December 2014, a total of 23 patients with Rockwood type III,Ⅳ,Ⅴshoulder joint dis-location were included, all cases was treated by arthroscopic double Endobutton fixaton technique.Patients were followed up for 6 to 18 months.The changes of shoulder joint forward elevation and external rotation, Japanese Orthopaedic Association score(JOA), and Visual Analogue score( VAS) of acromioclavicular joint in both perioperative and postoperative were recorded. Results The mean follow-up time was (9.3 ±2.4) months ( range from 6 to 18 months),and mean time from injury to surgery was(10.1 ±3.2)days.The therapeutic effects were achieved in all patients after the arthroscopically assisted double Endobutton plate reconstruction.All the measurements, in-cluding the mean forward elevation JOA scores before and after fixation (71.0 ±5.4 vs 93.5 ±3.6), and mean VAS scores (4.6 ±1.0 vs 1.9 ±0.9), mean forward elevation [(102.5 ±8.6) degrees vs (144.3 ±6.3) degrees], and mean external rotation[(107.1 ±6.2) de-grees vs (149.6 ±7.5) degrees] improved significantly (P<0.05) after the treatment. Conclusion Arthroscopic double Endobutton fixaton technique could provide an effective immobilization of shoulder and acromioclavicular joint, thus improving the function of the joint and achieving an excellent effect in the treatment of acute acromiocla-vicular joint dislocation.

Objective: To obtain monoclonal antibodies against putative protein X4 of SARS-CoV for further study of the structure and function of protein X4. Methods: Balb/c mice were immunized with recombinant Protein X4, hybridoma cell lines secreting monoclonal antibodies against protein X4 were screened by regular cell fusion and subcloning approach. The specificities of these monoclonal antibodies were determined by Western blotting and Immunofluorescecence assay. Results: Three hybridoma cell lines (8A5, 8H6 and 4A2) stable in secreting specific monoclonal antibodies were successfully obtained. They could bind specifically to protein X4 proved by Western blotting. All of them belonged to the IgG2a isotype proved by antigen mediated ELISA. Indirect Immunofluorescence assay indicated that they could specifically bind to protein X4 expressed in SARS-CoV infected Vero E6 cells. Conclusion: Monoclonal antibodies of high specificity against protein X4 have been successfully prepared, which laid the foundation for the further study of protein X4.

Objective To investigate the effects of early intensive therapy on P cell function and long-term glycemic control in newly diagnosed type 2 diabetic patients with different recruiting fasting plasma glucose (FPG) levels.Methods A total of 382 newly diagnosed type 2 diabetic patients with FPG 7.0-16.7 mmol/L were randomly assigned to therapy with insulin in the form of continuous subcutaneous insulin infusion (CSII) or multiple daily injection (MDI) or oral hypoglycemic agents (OHA, by using gliclazide and/or metformin) for initial rapid correction of hyperglycemia.The treatments were stopped after euglycemia had been maintained for 2 weeks.The patients were followed longitudinally on diet alone for 1 year.Intravenous glucose tolerances tests (IVCTTs) were performed and blood glucose, insulin and proinsulin were measured before and after therapy as well as at 1-year follow-up.Homeostasis model assessment ( HOMA) of β cell function and insulin resistance index ( HOMA-β and HOMA-IR ) were calculated.All the patients were stratified on the recruiting FPG: stratum A (7.0 mmol/L≤ FPG < 11.1 mmol/L) , stratum B (11.1 mmol/L≤ FPG ≤ 16.7 mmol/L).Results More patients in stratum A achieved target glycemic control (94.4% vs 89.8% ) and in shorter time [(5.9 ±3.8)d vs(6.9 ±3.6)d, P <0.05] as compared with those in stratum B.B cell function represented by HOMA-β and acute insulin response ( AIR) improved significantly after intensive interventions in both stratum A and B patients.However, the remission rate at 1 year was significantly higher in stratum A patients (47.8% ) than those in stratum B (35.7%, P < 0.05).The patients treated with insulin (especially with CSII) had higher remission rates and better improvement of AIR at 1 year follow-up irrespective of the recruiting FPG (CSII or MDI vs OHA: 57.1% , 51.8% vs 32.8% in stratum A, P <0.05; 44.4% , 38.7% vs 18.6% in stratum B, P <0.05).Conclusions Compared with OHA, early short time intensive insulin treatment had more favorable outcomes on maintaining AIR and prolonged glycemic remission in newly diagnosed type 2 diabetic patients irrespective of the recruiting FPG levels.

Objective:To investigate the influence of curative-intent resection with textbook outcomes of liver surgery (TOLS) on long-term prognosis of gallbladder carcinoma (GBC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 824 patients with GBC in the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, who were admitted to 15 medical centers from January 2014 to January 2021, were collected. There were 285 males and 539 females, aged (62±11)years. According to the evalua-tion criteria of TOLS, patients were divided into those who achieved TOLS and those who did not achieve TOLS. Measurement data with normal distribution were represented as Mean± SD, and com-parison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data were conduc-ted using the Mann-Whitney U test. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and the Log-rank test was used for survival analysis. The COX stepwise regression model with backward Wald method was used for univariate and multivariate analyses. Results:(1) Achievement of TOLS. Of the 824 patients undergoing curative-intent resection for GBC, there were 510 cases achieving TOLS and 314 cases not achieving TOLS. (2) Follow-up. Of the 824 patients undergoing curative-intent resection for GBC, after excluding 112 deaths within 90 days after discharge, 712 cases were included for the survival analysis. The median follow-up time, median overall survival time and 5-year overall survival rate of the 510 patients achieving TOLS were 22.1(11.4,30.1)months, 47.6(30.6,64.6)months and 47.5%. The median follow-up time, median overall survival time and 5-year overall survival rate of the 202 patients not achieving TOLS were 14.0(6.8,25.5)months, 24.3(20.0,28.6)months and 21.0%. There was a significant difference in overall survival between patients achieving TOLS and patients not achieving TOLS ( χ2=58.491, P<0.05). (3) Analysis of factors influencing prognosis of patients. Results of multivariate analysis showed that TOLS, carcinoembryonic antigen (CEA), CA19-9, poorly differentiation of tumor, T2 stage of eighth edition of American Joint Committee on Cancer (AJCC) staging, T3 and T4 stage of eighth edition of AJCC staging, N1 stage of the eighth edition of AJCC staging, N2 stage of the eighth edition of AJCC staging, adjuvant therapy were independent factors influencing overall survival time of patients undergoing curative-intent resection for GBC ( hazard ratio=0.452, 1.479, 1.373, 1.612, 1.455, 1.481, 1.835, 1.978, 0.538, 95% c onfidence interval as 0.352-0.581, 1.141-1.964, 1.052-1.791, 1.259-2.063, 1.102-1.920, 1.022-2.147, 1.380-2.441, 1.342-2.915, 0.382-0.758, P<0.05). Conclusion:Patients under-going curative-intent resection for GBC with TOLS can achieve better long-term prognosis.