BACKGROUND:Repeated injections or nasal spray of large doses of calcitonin can effectively prevent postmenopausal osteoporosis. Calcitonin should be taken for a long time.But the use of calcitonin is limited by the need for repeated protein administration, costly production methods and antigenicity. Gene therapy can provide effective economic therapeutic regimen for osteoporosis,and reduce side effect of drugs.OBJECTIVE:To describe the expression of human calcitonin produced in myoblasts and determine the effects of the recombinant protein on murine osteoblast cells.DESIGN:A gene-based controlled observational experiment.SETTING:Institute of Radiation Medicine,Fudan University.MATERIALS: The experiment was carried out at the Institute of Radiation Medicine, Fudan University from December 2005 to June 2006. Ten healthy SD fetal rats were selected from Institute of Radiation Medicine, Fudan University,Human Calcitoninsas monoclonal antibody was purchased from American Santa Cruz Biotechnology Company. L6 myoblast line was provided by Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences. METHODS:The pcDNA3.0.-hCT liposome transfection mixture (transfection group) and empty vector pcDNA3.0 liposome mixture (control group) were added in the L6 myoblast medium, respectively.The expression and secretion of human calcitonin by myoblast cells were confirmed by enzyme-linked immunosorbent assay (ELISA),Western blot analysis and immunohistochemical analysis.1×10-14,1×10-13,1×10-12 mol/L recombinant human calcitonin and MEM were respectively added in myoblast medium. MAIN OUTCOME MEASURES:The proliferation and differentiation of rat myoblasts were observed by MTT and alkaline phosphatase (ALP).RESULTS: Human calcitonin was found by ELISA in the supematant of cell culture. Western blot and immunohistochemical analysis verified that human calcitonin could be expressed stably in myoblasts after transfection. Osteoblast proliferation and ALP activity were higher when recombinant human calcitonin was 1×10-14 and 1×10-13 mol/L than the control group (P＞0.05).It was significantly higher when the concentration was 1×10-12 mol/L than the control group (P＜0.05).CONCLUSION:The stable synthesis and secretion of biologically active human calcitonin can be achieved in myoblasts by gene transfection.Recombinant human calcitonin can enhance proliferation and differentiation of osteoblasts.

Objective To explore influence of valsartan on MAU of patients with DN at early stage.Methods 163 patients with DN at early stage were randomly divided into experimental group and control group.Two groups were given routine therapy, and experimental group were added the therapy of valsartan.Period of treatment was 24 weeks, and compared the UAER, Cr, BUN of two groups.Results After treatment of 24 weeks, UAER, Cr, BUN of experimental groups was (142.4 ± 15.6) μg/min, ( 68.7 ±9.4) μmol/L, (3.5 ± 1.2 ) mmol/L, compared with (195.8±23.7)(μg/min,(93.8 ± 13.6) μmol/L, (6.3 ±1.5) mmol/L before treatment, had significant difference (t = 13.675,11.287,2.469,all P<0.05).Compared with( 199.6 ±24.7)μg/min,(87.7 ±11.3)μmol/L、(6.2 ± 1.3)mmol/L of control group after treatment, declined more significantly (t = 13.246,10.312,2.518,all P<0.05).There was no serious complications occurred in two groups.Conclusion Valsartan used in patients with DN at early stage,can significantly improve the function of kidney and enhance clinical effect,decline the side reaction.

Objective To study the effects of resin absorptive devices combined with hemodialyzer in series treatment for patients with uremia.Methods Sixty maintenance hemodialysis patients with uremia were randomly divided into two groups: the experimental group and control group. The patients in experimental group were given the treatment for traditional hemodialysis two or three times a week,combined with the treatment for resin absorption twice a month.The patients in the control group were only given the treatment for traditional hemodialysis two or three times a week.The middle molecular substance(MMS),beta(2) microglobulin(?_2-MG),blood urea nitrogen,creatinine,albumin,globulin,the counts of red blood,hemoglobin,the change of blood pressure and clinical manifestation of the patients were observed before and three months after the treatment respectively.Results The rates of clearance of MMS and ?_2-MG were significantly higher in the experimental group than those of the control group(P

Objective To investigate the effect of Bushenzhuanggutang combined with salmon calcitonin see calcimar in the treatment of renal osteodystrophy,and explore the mechanism of treatment.Methods 48 patients with renal osteodystrophy were divided into two groups:combined treatment group and control group.The patients were treated with Bushenzhuanggutang and salmon calcitonin see calcimar in combined treatment group.The patients were treated with salmon calcitonin see calcimar in the other group.Then the levels of serum calcium、phosphorus,PTH and BUN、Scr were detected before and after the treatment.Results The level of serum calcium increased significantly in two groups,while the level of serum phosphorus and PTH decreased(P

0.05) .It was significantly higher when the concentration was 1?10-12 mol/L than the control group(P

OBJECTIVE:To observe clinical efficacy and safety of salvianolate in the treatment of acute exacerbation of chron-ic obstructive pulmonary disease(AECOPD). METHODS:80 AECOPD patients were selected and randomly divided into observa-tion group and control group,with 40 cases in each group. Both groups received mechanical ventilation. Control group was given routine treatment of theophyllinum,bronchodilators and glucocorticoid;treatment group was additionally given Salvianolate injec-tion 200 mg,qd,ivgtt,on the basis of control group. Clinical efficacy was observed in 2 groups,blood hemorheology indexes were also observed before and after treatment. The mechanical ventilation time,ICU residence time and ADR were recorded in 2 groups. RESULTS:Clinical total effective rate of observation group was 95.00%,which was significantly higher than 70.00% of control group,with statistical significance(P<0.05). Before treatment and 2 d after treatment,blood rheology indexes of treatment group were improved significantly(all P>0.05),and whole blood reduction viscosity of control group was improved significantly(P<0.05). Mechanical ventilation time and ICU residence time of treatment group was significantly shorter than that of control group (P<0.05). No ADR was found in 2 groups. CONCLUSIONS:Salvianolate can significantly improve the blood coagulation status of AECOPD patients receiving invasive mechanical ventilation,and has the advantages of good clinical efficacy and low cost of medical treatment.

OBJECTIVE:To optimize the synthesis technology of 4-biphenylacetic acid,and to provide technology support to meet the market demand of 4-biphenylacetic acid raw material. METHODS:The synthesis route of 4-biphenylacetic acid from biphenyl by Friedel-Crafts reaction,ketal reaction,rearrangement reaction and hydrolysis was optimized and improved. The effects of different Friedel-Crafts reaction solvents (petroleum ether,dichloromethane,1,2-dichloroethane),ketal reaction ethanol (ethanediol, 1,3-propanediol, dimethyltrimethylene glycol, pentaerythritol), rearrangement reaction catalyzer (zinc biphenyl acetate,zinc oxide,zinc caprylate,zinc acetate) and refined solvent (isopropanol,95% isopropanol,ethanol,acetone,ethyl acetate) on synthesis technology were investigated. The optimal synthesis technology was screened with the purity and yield of intermediate or 4-biphenylacetic acid. RESULTS:The petroleum ether as Friedel-Crafts reaction solvent,ethylene glycol as ketal reaction ethanol,zinc diphenylacetate as rearrangement catalyst and 95% isopropanol as refining solvent were used for the preparation to obtain better effects. The yield of key intermediate 2′-chloroacetophenone reached above 95%. The purity of refined 4-biphenylacetic acid reached 99.9%;the content of single impurity was less than 0.1%;the total yield reached over 70%. CONCLUSIONS:The optimal synthesis technology of 4-biphenylacetic acid has the advantages of simple operation,mild and controllable reaction conditions,low cost,greener reagents and higher safety,and is suitable for industrial production. The purity and yield of the products are in high level and in line with the standards of European Pharmacopoeia and the Japanese Pharmacopoeia.

Objective: To explore the role of iTr35 cells in the pathogenesis of children with pulmonary artery hypertension (PAH) in children, and the percentage of iTr35 cells and serum interleukin(IL)-35 levels in peripheral blood of patients with PAH were investigated.Their inhibitory effects on the expression of vascular cell adhesion molecule-1 (VCAM-1) on vascular endothelial cells were also analyzed. Methods: After 3 mL peripheral blood of 30 congenital heart disease (CHD) patients with PAH, 22 CHD patients without PAH and 30 age or gender matched healthy controls (HC) were collected, the percentage of iTr35 cells were detected by flow cytometry and the concentrations of serum IL-35 were detected by Luminex, as well as soluble VCAM-1 (sVCAM-1). Human pulmonary artery endothelial cells (HPAECs) were cultured in vitro and divided into control group, tumor necrosis factor (TNF)-α group and IL-35+ TNF-α group.The expression of VCAM-1 and nuclear factor(NF)-κB P65 protein of each group were detected by flow cytometry and Western blot.The adhesion of peripheral blood mononuclear cells (PBMCs) to HPAECs was observed by fluorescence microscope. Results: Compared to CHD patients without PAH, the percentage of iTr35 cells[0.86(0.45-1.63)% vs.1.14(0.46-2.11)%](H=20.52, P<0.05) in peripheral blood and serum IL-35 levels[2.43(1.76-2.85) μg/L vs.3.17(2.92-5.66) μg/L](H=119.56, P<0.05) were significantly decreased in CHD patients with PAH.However, serum sVCAM-1 concentration[923.1(892.6-1 118.7) μg/L vs.776.1(743.5-932.3) μg/L ] in CHD patients with PAH were significant increased (H=65.65, P<0.05). In addition, the concentration of serum IL-35 were negatively correlated with sVCAM-1 in PAH patients (r=-0.374 P=0.042). In vitro, the positive rate of VCAM-1 on HPAECs was significant decreased in IL-35+ TNF-α group as compared to the TNF-α group [(2.07±0.82)% vs.(5.83±1.34)%, F=1 197.18, P<0.05]. In addition, the protein expression of NF-κB P65 in HPAECs was significantly decreased in TNF-α+ IL-35 group as compared to TNF-α group, as well as the adhesion of PBMCs to HPAECs (F=212.04, 2 533.51, all P<0.05). Conclusions The percentages of iTr35 and levels of IL-35 are reduced in the peripheral blood of patients with PAH.Through in vitro experiments, IL-35 is found to reduce PBMCs adhesion by inhibiting VCAM-1 expression in HPAECs.

Objective:To evaluate the effectiveness and prognosis of emergent transcatheter aortic valve replacement (TAVR) and to provide standardized procedural suggestion for the development of emergent TAVR in China.Methods:From January 2020 to April 2021, 12 patients who underwent emergent or salvage TAVR in the Second Affiliated Hospital Zhejiang University School of Medicine were retrospectively enrolled from the TORCH registry (Transcatheter Aortic Valve Replacement Single Center Registry in Chinese Population, a prospective cohort study; NCT02803294). Baseline, periprocedural and 30-day follow up data were collected. Post-operative data were compared with pre-operative data using Paired-Samples test.Results:Patients’ median Society of Thoracic Surgeons score (STS score) was 15.432%. There was a significant decrease of mean gradient after emergent TAVR procedure (1.69 m/s vs. 4.90 m/s, P<0.01). During the 30-day follow up, there were 1 patient (8.3%) died and 2 patients received permanent pacemaker implantation. No disabling stroke, acute kidney injury, major vascular complication occurred during the first month after emergent TAVR. Among the survival patients, there was a significant releasing of heart failure symptoms to New York Heart Association function stage Ⅰ/Ⅱ in 81.8% patients at 30-day follow up. Left ventricular ejection fraction also improved significantly from (47.4±9.5)% to 58.8±8.0% ( P= 0.026). The mean gradient were (1.57±0.30) cm 2 and no patients had a moderate or severe paravalvular leakage. Besides, a significant decrease of pro-B-type natriuretic peptide (1 089.9 pg/mL vs. 12 215.5 pg/mL , P=0.001) and troponin T (0.020 ng/mL vs. 0.337 ng/mL, P=0.003) were found at 30 days after emergent TAVR. Conclusions:For patients with severe aortic stenosis and acute cardiac decompensated, emergent TAVR is a safe and effective rescue treatment.

The incidence of intrahepatic cholangiocarcinoma (ICC) has been increasing year by year. For most patients, surgical resection is not suitable when they are diagnosed as ICC. Conventional chemotherapy and radiotherapy are not effective for the long-term survival rate of ICC patients and lead to the poor overall prognosis. In recent years, with the deepening understanding about the molecular mechanism of biliary malignant tumors, some key genes and signaling pathways related to the pathogenesis of ICC have been identified, providing new ideas for the targeted therapy. In this paper, major molecular mechanisms and targeted therapies of ICC are reviewed.