Objective To establish quality standard for Liangsang Diandiye. Method Radix et Rhizoma Notoginseng, Flos Carthami and Radix Salviae Miltiorrhiizae were identified by TLC. Ginsenoside Rg1 was determined by dual-wavelength TLC-scanning with 525 nm and 700 nm as the detective wavelength. Results Radix et Rhizoma Notoginseng, Flos Carthami and Radix Salviae Miltiorrhiizae can be identified by TLC. A good line relationship was noted at the range of 1.53～7.65 ?g Ginsenoside Rg1 (r =0.9991). The average recovery was 99.10%, and RSD was 0.27%. Conclusion The established method can be used for the quality control of Liangsang Diandiye.

Objective To investigate the protective effect of butylphthalide for focal cerebral ischemiareperfusion injury and its possible mechanism.Methods A total of 60 healthy and clean adult Sprague-Dawley rats were randomly divided into sham operation,saline control,low-dose butylphthalide and high-dose butylphthalide groups (n =15 in each group).A focal cerebral ischemia-reperfusion model was induced by the suture method.At the beginning of reperfusion,100 rng/kg and 400 mg/kg butylphthalide injection were injected intraperitoneally in the rats of the low-dose butylphthalide and high-dose butylphthalide groups; 0.5 ml/kg saline was injected intraperitoneally in rats of the sham operation and saline control groups.All the rats were sacrificed after 24 h of ischemia-reperfusion.Results The degree of neurological deficit in the low-dose butylphthalide (t =1.488,P =0.000) and high-dose butylphthalide (t =2.362,P =0.000) groups were significantly improved compared to the saline control group,in which the high-dose butylphthalide group was improved more significantly than the low-dose butylphthalide group (t =-0.873,P =0.000).The infarct volume in the low-dose butylphthalide (t =18.589,P =0.000) and high-dose butylphthalide (t =36.963,P =0.000) groups were reduced significantly compared to the saline control group,in which the infarct volume of the high-dose butylphthalide group was reduce more significantly than that of the low-dose butylphthalide group (t =-18.374,P =0.000).HE staining showed that neurons were sparse,there were a large number of degeneration and necrosis,cell space became larger,and the intercellular substances showed vacuolar changes.In the butylphthalide group,the neuronal degeneration and necrosis reduced significantly,the survival of nerve cells increased,and the improvement of the high-dose butylphthalide group was more remarkable.SOD activity of the low-dose and high-dose butylphthalide groups were increased significantly compared to the sham operation and saline control groups (all P ＜0.05),in which the SOD activity in the high-dose butylphthalide group was significantly higher than that in the low-mall dose butylphthalide group (t =80.199,P =0.000); The MDA levels in the low-dose and high-dose butylphthalide groups were decreased significantly compared to the sham operation and saline control groups (all P ＜ 0.05),in which the MDA level in the high-dose butylphthalide group was significantly lower than that in the low-dose butylphthalide group (t =-1.308,P=0.000).Conclusions The protective effect of butylphthalide on ischemia-reperfusion injury may be associated with the increased antioxidant activity.

OBJECTIVE To improve efficiency and quality of antibiotic drug usage in order to assure patients′ security of antibiotic drug usage. METHODS Inducted by strengthening security quality of medical treatment,we are bring antibiotic drug usage into medical quality management by using information technique,training medical workers,putting antibiotic drug into different classifications and surveillancing the usage of antibiotic drug.Moreover,we develop significant basic study for clinic to improve the level of rational use of drug. RESULTS Frequency and number of days for drug usage were reduced by putting measures into effect and inspecting numbers and sorts of drug usage.At the same time,we instituted individualized drug using scheme to critical patients,thereby many critical patients were retrieved. CONCLUSIONS Rational use of drug is a system project which must redeploy all enthusiasm and take comprehensive measures to fight for a safe,effective,economical goal in our work.

Aim Myocardial cells of neonatal rats were cultured in vitro to study aim of morphine on serum hungry-induced apoptosis in cardiac myocytes and its mechanism.Methods Myocardial cells of neonatalrats were cultured in vitro.48 hours later,different agents were added to cardiac myocytes.The cellular survival was determined with MTT colormeteric assay;apoptosis rates were determined by Annexin V-FITC/PI;cell cycle was determined by flow cytometry;Caspase-3 and PKC were investigated by Western blot.Results Free-serum induced apoptosis in cardiac myocytes was shown after 48 hours;morphine(1 ?mol?L-1)could inhibit apoptosis in cardiac myocytes,manifestation,apoptosis rates were decreased,Caspase-3 experssion was decaeased Naloxone at 10 ?mol?L-1 inhibited the promoting effects of morphine;GF109203X at 10 ?mol?L-1 or Staurosporine at 1 ?mol?L-1 could inhibit the promoting effects of morphine,manifestation,apoptosis rates were increased,Caspase-3 experssion was incaeased,PKC expression was decreased.Conclusion morphine inhibited serum hungry-induced apoptosis in cardiac myocytes via PKC signal transduction pathway.

Aim To investigate the possible mechanism of ?-opioid receptor inhibiting serum deprivation-induced apoptosis of neonatal cardiac myocytes.Methods Myocardial cells of neonatal rats were cultured in vitro,after 48 h,the medium was changed to serum-free DMEM.The experimental groups were:A:Normal;B: Model;C: Model+DADLE(0.1 ?mol?L-1);D:Model+DADLE(0.1 ?mol?L-1)+naltrindole(10 ?mol?L-1);E:Model+DADLE(0.1 ?mol?L-1)+GF109203X(10 ?mol?L-1);F:Model+DADLE(0.1 ?mol?L-1)+staurosporine(1 ?mol?L-1).The cell viability was determined with MTT colormeteric assay;apoptosis index and the percentage of G0 in cell cycle were determined by flow cytometry;Caspase-3 and PKC signal pathway was investigated by Western blot.Results ?-opioid receptor agonist [D-Ala2,D-Leu5]-enkephalin(DADLE) could significantly inhibit serum deprivationinduced apoptosis of neonatal cardiac myocytes,which increased the survival index of cardiac myocyte,percentage of G2/M in cell cycle and the expression of PKC,decreased the apoptotic index of cardiac myocyte,percentage of G0/G1 in cell cycle and the expression of activate caspase-3.The protective effect of DADLE was obviously blocked by ?-opioid receptor antagonist Naltrindole at 10 ?mol?L-1,the PKC inhibitor GF109203X at 10 ?mol?L-1 and staurosporine at 1 ?mol?L-1,which decreased the survival index of cardiac myocyte,percentage of G2/M in cell cycle and the expression of PKC,increased the apoptotic index of cardiac myocyte,percentage of G0/G1 in cell cycle and the expression of caspase-3.Conclusion These findings suggested that ?-opioid receptor activation might be a potential survival factor against serum deprivation-induced myocardial cell apoptosis and this cardioprotective effect might be via PKC pathway.

Objective To investigate the therapeutic effects of recombinant adeno-associated virual gene serotype 1(rAAV1) mediated transfer of sarcoplasmic reticulum Ca2+ ATPase(SERCA2a) on beagle dogs with heart failure(HF).Methods The chronic HF model was reproduced in beagle dogs by giving rapid right ventricular pacing(230 beats/min) for 30 days.A reduced rate(180 beats/min)was continued for another 30 days.Sixteen beagle dogs were divided into four groups(4 each): control group,HF group,HF+EGFP group and HF+SERCA2a group.After rapid pacing for 30 days,rAAV1-EGFP(1?1012vg/ml) and rAAV1-SERCA2a(1?1012vg/ml) were respectively delivered via intramyocardial routes,while no treatment was given to the animals in both control and HF groups.At the end of the study,haemodynamics,echocardiography and the protein expression of SRCA2a were measured respectively.The apoptosis index of cardiac myocyte was evaluated by TUNEL.Bax expression was assessed by immunohistochemistry.Results After gene transfer of SERCA2a in HF beagle dogs for 30 days,the heart function was improved along with an increase in SERCA2a expression.Left ventricular systolic function was significantly increased,including the left ventricular systolic pressure(LVSP),left ventricular maximal rate of pressure rise(LV+dp/dtmax),left ventricular maximal rate of pressure decline(LV-dp/dtmax,P

Objective To determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of chemokines and nuclear factor-κB (NF-κB) signaling pathway. Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. The cells were divided into control group, LPS challenge group, 1 kU/L or 10 kU/L UFH+LPS group, and NF-κB inhibitor N-tosyl-L-lysyl chloromethyl-ketone (TLCK) group (TLCK+LPS group). HPMECs in LPS challenge group were treated with 10 mg/L LPS. UFH pretreatment with different dosages groups were treated with 1 kU/L or 10 kU/L UFH 15 minutes before LPS challenge. Cells in the TLCK+LPS group were treated with 10 μmol/L of TLCK 30 minutes before the addition of LPS, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The cells were harvested 1 hour after LPS challenge, and the nuclear translocation of NF-κB was determined by immunofluorescence assay to detect the effect of UFH on NF-κB activation. The levels of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 3 hours and 6 hours after LPS challenge to detect the effect of UFH on LPS induced expression of chemokines and its mechanism of effect on NF-κB signaling pathway in HPMECs. Results ① In the control group, NF-κB was mostly located in the cytosol as shown by immunofluorescence. Treatment of HPMECs with LPS significantly increased the translocation of NF-κB from the cytosol to nucleus. UFH suppressed LPS-induced NF-κB activation both in 1 kU/L and 10 kU/L dosages, and 10 kU/L UFH gave even better results. ② Compared with control group, the levels of IL-8 and MCP-1 in the supernatants in LPS challenge group were significantly increased at 3 hours and 6 hours after LPS challenge [IL-8 (ng/L): 387.1±26.4 vs. 23.8±8.1 at 3 hours, 645.5±69.6 vs. 125.7±18.7 at 6 hours; MCP-1 (ng/L): 3 654.9±467.9 vs. 721.6±61.3 at 3 hours, 8 178.5±792.6 vs. 1 324.7±148.7 at 6 hours, all P < 0.05]. Compared with that of LPS challenge group, in 1 kU/L and 10 kU/L UFH pretreatment groups, the levels of IL-8 and MCP-1 were significantly decreased [IL-8 (ng/L): 315.3±24.8, 275.8±31.1 vs. 387.1±26.4 at 3 hours, 557.8±43.3, 496.9±38.7 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 2 924.1±267.9, 2 668.3±522.6 vs. 3 654.9±467.9 at 3 hours, 7 121.7±557.2, 6 563.9±576.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. The results indicated that 10 kU/L UFH yielded better results. However, inhibition study using the known NF-κB inhibitor TLCK could decrease LPS-induced increase in IL-8 and MCP-1 levels [IL-8 (ng/L): 162.4±21.3 vs. 387.1±26.4 at 3 hours, 274.1±22.6 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 1 478.2±138.5 vs. 3 654.9±467.9 at 3 hours; 3 667.6±259.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. Conclusions The levels of IL-8 and MCP-1 were increased obviously in LPS treated HPMECs. UFH might suppress LPS-activated NF-κB signaling pathway, contributing to the inhibitory effects of chemokines in HPMECs.

Objective To evaluate the detection competence of HbA2 and HbF in Guangxi medical laboratories.Methods The external quality assessment(EQA) of HbA2 and HbF was conducted twice a year and five samples was detected each time during 2012 to 2016.The laboratories participated in EQA completed the samples' detection and submitted the detection results at specified time according to the requirements of EQA.The distribution of each detection system,the qualification rate of each laboratory,the variation degrees of each detection system and each detection method,and the variations of results for different levels of quality control(QC) materials during 5 years were analyzed based on the returned results.Results The application of high performance liquid chromatography (HPLC) and capillary electrophoresis(CE) increased year by year and their usage rates in 2016 reached up to 46.1％ (82/178) and 18.0％ (32/178),respectively.The qualification rates of HbA2 and HbF increased from 51.5％ (34/66) and 60.6％ (40/66) in 2012 to 93.3％ (166/178) and 92.1％ (164/178) in 2016,respectively.The average coefficient of variation(CV) of each detection system decreased year by year.There were good CVs for the results of high,medium and low levels of HbA2 QC materials detected by the Bio-Rad Variant Ⅱ and Sebia CAPILLARYS 2 systems,and they were less than 6.0％.HPLC and CE could quantitatively detect the HbA2 and HbF levels,and their total detection competence was superior to that of agarose gel electrophoresis.Conclusion EQA can assess the abihty of one laboratory detecting HbA2 and HbF,and quantitatively analyze the levels of HbA2 and HbF,which may provide the quality assurance and data support for the screening and prevention of thalassemia.

BACKGROUND: Bone morphogenetic protein (BMP) is a protein possesses potential activity, which can increase and enhance its activity when the bone issues are damaged, so it can be used to repair the bone defects when combined with carrier. However,there are few reports concerning it as gene therapy.OBJECTIVE: To construct recombinant retroviral vector expressing human BMP2 gene and to discuss its biological function in ostecblasts.METHODS: BMP2-specific primers were designed and synthesized according gene sequence of human BMP2 gene in Genbank, then BMP2 gena was amplified by Hifi PCR, which was recombined with cloning vector pDNR-CMV homologousiy into pDNR-CMV-BMP2 plasmid identified by BMP2 PCR and enzyme digestion of Sail and EcoRI as well as gene sequencing; recombinant plasmid pDNR-CMV-BMP2 and retroviral plasmid pLP-LNCX were recombined homologously in IoxP sites into pLP-LNCX-BMP2 plasmid transferred into packing cell line PT67 and the supernatant was collected to assay viral titre. Human osteoblast was infected with retrovirus, then the growth of cells were observed by MTT, and the expression of BMP2 protein was detected by Western blotting at 48 hours transfectionRESULTS AND CONCLUSION: Digestion, BMP2 PCR and gene sequencing of pDNR-CMV-BMP2 were coincided with expected. After transfected with plasmid pLP-LNCX-BMP2, PT67 cells could be screened with G418 only to get stably integrated in BMP2, of whose supemanant viral titra amounted to 5×10~8 pfu/mL. MTT assay showed that there had no evident difference in cellular inhibition between the normal and retrovirus groups at 72 hours after transfection (P ＞ 0.05); Western blotting showed that the BMP2 was strong expressed at 48 hours after transfection. It demonstrated that BMP2 gene was successful cloned and its retrovirus vector was constructed.

Objective:To study the effects of cyclosporine A coadministrated with azathioprine,mycophenolate, mizorihine,rapamycin and/or prednisone on liver function in renal transplant recipients.Method:The drug history records of 600 renal transplant recipients in 1995 to 2005 were retrospectively investigated.Biochemical indexes before and after the treatment with cyclosporine A coadministrated with other immunosuppressants were analyzed.Result:The liver damage was found in 109 cases(18.2%)among 600 cases.The blood concentrations of cyclosporine A in the group with abnormal liver functions were significantly higher than those in the group with normal liver functions(P