Objective To establish a microbial limit test for levofloxacin hydrochloride gel. Methods Ca2+ was added to neutralize the antibacterial activity of quinolone,and to form the insoluble precipitate by binding with carbomer, therefore destroying the stability of macromolecules in the gel. The microbial limit test for levofloxacin hydrochloride gel was confirmed combining with membrane filtration and plate methods. Results The normal plate counting was used in detecting the mold and yeast colony in the gel, the recovery was more than 70%.Simultaneously neutralization centrifugation and membrane filtration were used in bacterial count,with recovery less than 70%,and controlling bacteria test. Conclusion The method is accurate and practical for microbial limit test of levofloxacin hydrochloride gel, but is risky for bacterial counting and needs further risk assessment.

OBJECTIVE: To observe the clinical effect of cinobufacini injection in treating moderate and advanced primary liver cancer (PLC). METHODS: One hundred patients with moderate and advanced PLC were randomly divided into cino-treated group (50 patients) and control group (50 patients). The quality of life, tumor size, some changes of laboratory tests, and survival time were observed. RESULTS: The progressive rate of cino-treated group (18%) was lower than that of the control group (32%). The quality of life of the cino-treated group (80%) was better than that of the control group (72%), but without statistical significance. The survival rate of >12 months of the cino-treated group (30%) was higher than that of the control group (18%). The patients' liver function such as serum total bilirubin and ALT decreased obviously in the cino-treated group while increased a lot in the control group. The level of AFP increased after treatment with statistical significance in the control group while there was no statistical significance in the cino-treated group. CONCLUSION: Cinobufacini injection can not only inhibit the proliferation of cancer, but also protect liver function, improve quality of life and prolong survival time.

Objective To evaluate the correlation between left ventricular function and arterial stiffness of left femoral artery in patients with lower extremity atherosclerotic disease (LEAD).Methods Thirty-three patients with LEAD and 37 healthy subjects (control group) were enrolled in this study.The intima-media thickness (IMT),diameter and parameters of arterial stiffness [dispensability coefficient (DC),compliance coefficient (CC),stiffness α,stiffness β,pulse wave velocity (PWVβ) ]were measured by ultrasonography with the technology of QIMT and QAS.The thickness of the interventricular septum (IVSd),end-diastolic left ventricular diameter (LVDd) and left ventricular mass (LVM),and parameters of the left ventriculsr function (EF,E/A,E'/A',E/E' and Tei index) were measured by echocardiography.These parameters were compared between two groups.Correlations between the parameters of the arterial stiffness and those of the cardiac function were evaluated by Pearson correlative analysis.Results ①The IVSd,LVM and E/E' ratio were significantly higher in LEAD group than those in control group ( P ＜0.05).There were no significant differences in EF,E/A,E'/A',and Tei index between two groups ( P ＞0.05).②The IMT,α,β,PWVβ of left femoral artery were significantly higher in LEAD group than those in control group,while DC and CC were significantly lower in LEAD group than those in control group ( P ＜0.05).③The E/E' ratio,one of the parameters representing the left ventricular diastolic function,was correlated negatively with CC and positively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).The E'/A' ratio was correlated positively with DC and CC,and negatively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).Both EF and Tei index were not significantly correlated with the above parameters of arterial stiffness ( P ＞0.05).Conclusions Patients with LEAD have thickened femoral IMT,higher arterial stiffness of left femoral artery,as well as impaired left ventricular function.There is a close correlation between the atherosclerosis of the femoral artery and the early left vcntricular dysfunction.

Objective To evaluate the association between the left carotid arterial stiffness and left ventricular diastolic function in patients with lower extremity atherosclerosis (AS).Methods ①A total of 32 patients with AS and 34 control objects were enrolled.The carotid arterial stiffness parameters:compliance coefficient (CC),distensibility coefficient (DC),stiffness parameter (α,β),pulse wave velocity β (PWVβ) were measured by using quality arterial stiffness(QAS) technology.And the values were compared between the two groups.②The parameters of left ventricular (LV) structure and function:LV end-diastolic interventricular septal thickness (IVSd),LV end-diastolic diameter (LVDd),LV end-diastolic wall thickness (PWd),LV ejection fraction (EF),systolic velocity (s'),early-diastolic velocity (e'),Tei index and E/e' ratio were measured by using two-dimensional echocardiography and tissue Doppler.These parameters were compared between the two groups.The association between the carotid arterial stiffness parameters and LV function parameters were analyzed by correlative analysis.Results ①Compared with the control group,the DC and CC were lower,and α,β,PWVβ,IMT were higer than the control group,with statistically significant differences(P ＜0.05).②The IVSd,Tei index and E/e'was significantly higher in the AS group than those in the control group.And the PWd,s',e' were lower than those in the control group (P ＜ 0.05).There was no significant difference in EF between the two groups (P ＞0.05).③The e' was correlated positively with DC and CC (r =0.39,0.36,P ＜0.01),and negatively with α,β,and PWVβ (r =-0.42,-0.42,-0.49,P ＜0.01).Tei index was correlated negatively with DC and CC (r =-0.50,-0.52,P ＜0.01),and positively with α,β,and PWVβ (r =0.58,0.58,0.62,P ＜0.01).The E/e' was correlated regatively with CC (r =-0.27,P ＜0.05),and positively with PWVβ (r =0.28,P ＜0.05).There were no significant correlation between s',EF and the stiffness parameters of carotid artery (P＞0.05).Conclusions In patients with AS,the left carotid artery stiffness increases and left ventricular systolic and diastolic function are impaired.The carotid artery stiffness and left ventricular diastolic function is correlated.Changes in carotid artery stiffness reflect the change in left ventricular diastolic function.

Animals ; Anticholesteremic Agents ; pharmacology ; Cholesterol ; blood ; Drug Evaluation, Preclinical ; methods ; Male ; Rats

Objective To summarize the technique and managements of complications in endovascular embolization on intracranial aneurysm with Guglielmi detachable coils(GDC),and to evaluate the effect of the treatment.Methods One hundred and thirty six cases with Aneurizym were treated using GDC to embolize the aneurismal sac via femoral artery approach.Results One hundred and thirty six aneurysms were cured.Of them 132 cases recovered clinically,4 patients died.The mortality was 2.9%. The sac of 123 aneurysms were embolizied at 100%,8 cases with 95% embolization,5 with 90% embolization.3 aneurysms reptured during the embolization,cerebral vasospasm happened in 7 eases. microcoil escaped in 2 case.Three recurring cases were cured after second GDC embolization.The technique-related complications occured in 13 cases.No re-bleeding occurred during the 6 to 54-month follow-up.Conclusion Endovascular embolization of intracranial aneurysm with coils is a safe and effective treatment for intracranial aneurysm.Advances in Techniques and treating the complications correctly would decrease the complications and improve future outcomes.

Objective To construct recombinant expression vector TAT-SOX2 and express the fusion protein in E.coli BL21.Methods SOX2 fragment were amplified by PCR.The product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contains protein transduction domain of TAT.The recombinant vector was transformed into E.coli BL and induced with IPTG.The expressed fusion protein was analyzed by using SDS-PAGE method.The highly expressed product was purified by affinity chromatography.Western blotting was used to identify the specificity of the fusion protein.The immunofluorescence was used to detect the efficiency of fusion protein transduced to HSF cells.Results The recombinant expression vector TAT-SOX2 was correctly constructed and fusion protein TAT-SOX2 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein was transduced into HSF cells quickly.Conclusion This study provides the material basis for the further induced pluripotent stem cells by protein transduction.

Objective To construct recombinant expression vector TAT-KLF4 and express the Fusion Protein in E.coli BL21.Methods The open reading frame(ORF) of KLF4 gene was amplified by PCR.The PCR product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contained protein transduct domain of TAT.The recombinant vector was transfected into E.coli BL21 and the expression of the TAT-KLF4 fusion protein was induced with IPTG.The fusion protein was analyzed by using SDS-PAGE and purified by the affinity chromatography.Western Blot was used to identify the specificity of the fusion protein.The immunofluorescence was used to detection the efficiency of fusion protein transducted to HSF cells.Results The recombinant expression vector TAT-KLF4 was correctly constructed and fusion Proteion TAT-KLF4 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein transduced into HSF cells quickly.Conclusion This study provides the material basis for further induction of the induced pluripotent stem cells by protein transduction.

OBJECTIVETo study the effect of geniposide-acid(GA) on the anti-inflammatory action for adjuvant-induced arthritis (AA) rats and the proliferation of synoviocytes in AA rats and the feasible mechanism of apoptosis in vitro.

METHODForty-eight health male Wistar rats were divided randomly into six groups and were administered respectively with 200, 100, 50 mg x kg(-1) GA and 0.75 mg x kg(-1) MTX and normal sodium (normal or model control group) for four weeks when right posterior paw pads of rats excluding normal control group were injected intrademally with complete Freund's adjuvant after 19 days. The left posterior paws swelling degree, swelling inhibition ratio and arthritis index of secondary inflamation were detected. The TNF-alpha and IL-1beta proteins in serum of rats were assayed by enzyme linked immunosorbent assay (ELISA) kits. The synovial fibroblasts of AA rats were exposed to 1-4 micromol x L(-1) GA or 4 micromol x L(-1) MTX. The effect of GA on the proliferation of synoviocytes was detected by MTT assay. The morphologic change of apoptosis cells was observed by Hoechst/PI double stainning and fluorescence microscope. The rate of apoptosis cells was analyzed by flow cytometry. The mRNA expresstion of Bcl-2 and Bax gene was detected by reverse transcription PCR (RT-PCR).

RESULT200 mg kg(-1) or 100 mg kg(-1) GA could decrease significantly the paw swelling degree, arthritis index and the level of TNF-alpha and IL-1beta proteins in serum of AA rats (P < 0.05 or P < 0.01) with 25.4%, 21.37% of the swelling inhibition ratio respectivly, 34.61%, 28% of protein inhibition ratio of TNF-alpha and 29.05%, 21.65% of that of IL-1beta. GA(1-4 micromol x L(-1)) inhibitated significantly the proliferation of synoviocytes culcured for 5 days. Flow cytometry showed that 1, 2, 4 micromol x L(-1) GA increased obviously the rate of apoptosis cells, the apoptosis ratios were 15.8%, 24.3%, 40.7% respectivly (P < 0.01). RT-PCR showed GA could decrease the expression level of Bcl-2 gene but increase that of Bax gene (P < 0.05 or P < 0.01).

CONCLUSIONGA could inhibit the secondary inflamation of AA rats and decrease the level of TNF-alpha and IL-1beta protein in the AA rats serum. GA could inhibit the proliferation of AA rat synoviocytes in vitro and induce apoptosis which mechanism was concerned with down-regulating the mRNA expression of Bcl-2 and up-regulating that of Bax.

Animals ; Anti-Inflammatory Agents ; administration & dosage ; Apoptosis ; drug effects ; Arthritis, Experimental ; chemically induced ; drug therapy ; immunology ; physiopathology ; Cytokines ; immunology ; Disease Models, Animal ; Freund's Adjuvant ; adverse effects ; Glucosides ; administration & dosage ; Humans ; Iridoid Glucosides ; Iridoids ; administration & dosage ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Synovial Fluid ; cytology ; drug effects ; immunology

Aim To observe the effect of laminarin L01 on the expression of eNOS and iNOS in aorta of rats with chronic inflammation induced by LPS. Methods Chronic inflammatory rat models were prepared by tail vein injection low dose LPS(0.4 mg·kg-1) once a week for four weeks. The rats were randomly divided into five groups. After the first injection of LPS, the DXM group was intraperitoneally injected with dexam-ethasone (10 mg·kg-1). L01 high,medium and low dose groups were intraperitoneally injected with L01 (50,30,10 mg·kg-1). The LPS group was injected intraperitoneally with equal volume of normal saline once a day. Another control group, only injection of normal saline, a total of four weeks. After the last administration,the number of whole white blood cells (WBC) was counted. ELISA was used to measure the hs-CRP in serum. The expressions of eNOS,iNOS and COX-2 mRNA were detected by RT-PCR. Results After four weeks of administration of L01, the number of WBC and the level of serum hs-CRP in chronic in-flammatory rats were significantly decreased. The ex-pression of eNOS was up-regulated, and iNOS and COX-2 expressions were down-regulated. Conclusions Laminarin L01 may regulate the expression and re-lease of endothelium-derived relaxing factor stimulated by LPS,and improve the endothelium-dependent dias-tolic function of aorta, thus protecting the damage of vascular endothelium.