Objective To explore the risk factors of progressive cerebral infraction.Methods 112 cases with progressive cerebral infarction and 112 cases of acute cerebral infarction were selected.Patients were observed and recorded blood pressure,temperature,WBC,cholesterol,GLU and fibrinogen.Results The differences between blood pressure,temperature,WBC,cholesterol,GLU,fibrinogen and artery stenosis were statistically significant.Conclusion Infections,systolic blood pressure decreased,hyperglycemia,hyperlipidemia,and large artery stenosis were related to progressive cerebral infarction.

Objective To evaluate immunogenicity of the recombinant protein GST-p55/570 of Pneumocystis carinii.Methods The fusion protein GST-p55/570 was expressed from the prokaryotic expression plasmid pGEX-570,and purified by using glutathione-agarose.The expressed product was analyzed by SDS-PAGE.Thirty-three mice were randomly divided into three groups,immunized with GST-p55/570,GST and PBS,respectively.Each group was immunized for four times at 2 week intervals.At the 7th day after final immunization,spleen was removed to obtain single cell suspension.Proliferation ability of lymphocytes was determined by MTT.Serum samples were collected at pre-immunizaton and two weeks after each immunization.Antibody level in sera of mice was determined by ELISA.The immune response to the recombinant GST-p55/570 recognized by sera of immunized mice was examined by Western blotting.Results The expressed fusion protein GST-p55/570 showed a Mr 47 000.Compared with GST group(1.134 5?0.073 5) or PBS group(1.124 8?0.041 6),a higher stimulation index(2.063 0?0.160 2) was revealed in GST-p55/570-immunized mice(P

A method for the determination of methylmercury in seafood has been developed using dispersive liquid-liquid microextraction followed by direct mercury analyzer. Total mercury was detected by direct mercury analyzer, and inorganic mercury was calculated by the difference. The parameters affecting the extraction efficiency, including the selection of extractant and dispersant, their volume ratio, concentration of HCl and NaCl have been optimized in this study. The results showed that CH2 Cl2 as extractant, ethanol as dispersant, Volume ration of 1:5, 1 mol/L HCl and 120 g/L NaCl were chosen. The detection limit and the dynamic liner range were 0. 10 μg/L and 0. 2-20 μg/L, respectively. The relative standard deviation was 6. 0% for eleven replicates at the spiked level of 2. 0 μg/L. The enrichment factor was 8. For total Hg determination, the detection limit and the dynamic liner range for methylmercury were 0. 10 μg/kg and 0. 2-50 μg/kg, respectively. The relative standard deviation was 2. 4%. The method was simple, fast and a little solvent needed. Some certified reference materials were analyzed to validate the accuracy of the proposed method, and the results were in good agreement with the reference value. Besides, the method was applied to the real samples with satisfactory results.

To construct the eukaryotic expression plasmid containing the p55 gene fragment of Pneumocystis and to investigate the efficient expression in COS-7 cells, the gene fragment conaining the whole length of p55 gene was used as template to amplify this fragment with PCR and the amplified fragment was then cloned to vector pGEM-T. After enzyme digestion, p55 gene was cloned to the eukaryotic expression vector pcDNA3.1(+) to construct the plasmid pcDNA3.1(+)-582. This plasmid was then transfected to the eukaryotic expression cells COS-7 and PCR and SDS-PAGE assays were used to confirm the presence of target protein in these cells. In these ways, the eukaryotic expression vector for the p55 gene of Pneumocystis of rats was successfully constructed and expressed in COS-7 cells, thus providing the basis for further studies on the nucleic acid vaccine.

This study examined endogenous cannabinoid (ECB)-anandamide (AEA) and its cannabinoid receptors (CBR) in mice liver with the development of schistosoma japonicum. Mice were infected with schistosoma by means of pasting the cercaria onto their abdomens. Liver fibrosis was pathologically confirmed nine weeks after the infection. High performance liquid chromatography (HPLC) was employed to determine the concentration of AEA in the plasma of mice. Immunofluorescence was used to detect the expression of CBR1 and CBR2 in liver tissue. Morphological examination showed typical pathological changes, with worm tubercles of schistosoma deposited in the liver tissue, fibrosis around the worm tubercles and infiltration or soakage of inflammatory cells. Also, CBR1 and CBR2 were present in hepatocytes and hepatic sinusoids of the two groups, but they were obviously enhanced in the schistosoma-infected mice. However, the average optical density of CBR1 in the negative control and fibrosis group was 13.28+/-7.32 and 30.55+/-7.78, and CBR2 were 28.13+/-6.42 and 52.29+/-4.24 (P<0.05). The levels of AEA in the fibrosis group were significantly increased as compared with those of the control group. The concentrations of AEA were (0.37+/-0.07) and (5.67+/-1.34) ng/mL (P<0.05). It is concluded that the expression of endocannabinoids AEA and its cannabinoid receptor CBR were significantly increased in schistosoma-infected mice. Endogenous endocannabinoids may be involved in the development of schistosoma-induced liver fibrosis.
Arachidonic Acids/*metabolism ; Endocannabinoids/*metabolism ; Liver Cirrhosis/etiology ; Liver Cirrhosis/*metabolism ; Liver Cirrhosis/parasitology ; Polyunsaturated Alkamides/*metabolism ; Random Allocation ; Receptor, Cannabinoid, CB1/*metabolism ; Receptor, Cannabinoid, CB2/*metabolism ; Schistosomiasis japonica/*complications ; Schistosomiasis japonica/metabolism

Objective To investigate the changes and clinical correlation of IL-4 in patients with HBV infection.Methods Sixty cases with chronic asymptomatic HBV carriers,60 cases with chronic hepatitis B,60 cases with liver cirrhosis,60 cases with hepato-cellular carcinoma and 50 healthy controls were collected for serum.ELISA was used for detection of cytokine IL-4 levels;The a-mount of HBV DNA was measured by fluorescence quantitative PCR;Liver function was tested by automatic biochemical analyzer. Results Compared with the healthy controls[(1.64±0.17)ng/mL],IL-4 levels of patients with CHB,LC and HCC were signifi-cantly increased[(4.18±0.48),(4.71±0.42),(3.62±0.31)ng/mL,P <0.05].LC group have the highest IL-4 levels,while ASC was the lowest.Compared with the ASC group,IL-4 levels of patients with CHB,LC and HCC were significantly increased(P <0.05).Compared with the HCC group,IL-4 levels of patients with LC were significantly increased(P <0.05).IL-4 level was posi-tively correlated with TBIL levels in LC patients(r=0.529,P <0.01),while the IL-4 level was positively correlated with ALT and TBIL level in HCC patients(r=0.263,0.323,P <0.05).Conclusion IL-4 may play an important role in chronic HBV infection, the levels of IL-4 can be used as an important indicator to assess the severity of chronic hepatitis B.

Objective To study the effects of medicated serum with total saponins from Rhizoma Dioscreae Nipponicae (RDN) on VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α. To investigate the mechanism about total saponin from RDN inhibition of angiogenesis. Methods Medicated serum of total saponins from RDN and tripterygium (positive control) were prepared. Rat synovial cells RSC-364 were divided into four groups: the blank control,IL-17+TNF-α model,tripterygium medicated serum,and total saponins medicated serum groups. After one hour of incubation,all groups except for the blank control were incubated with both IL-17(10 μg·L-1 ) and TNF-α(10 μg·L-1 ) for 24 hours. VEGF mRNA expression in RSC-364 was detected by PrimeScriptTM real-time quantitative PCR (RT-PCR) detection kit,and the AP-1 DNA-binding activity was detected by electrophoretic mobility shift assay (EMSA). Results Compared with the control blank group,both of the VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α increased remarkably (P<0. 05,P<0.01). The VEGF mRNA expression and AP-1 activity in tripterygium medicated serum group and total saponins medicated serum group were remarkably lower than those of the model control group (P<0.05). There was no significant difference between the two medicated serum groups. Conclusion Serum medicated with total saponins from RDN can remarkably decrease VEGF mRNA expression and AP-1 activity,indicating that the total saponins from RDN could influence VEGF secretion by inhibiting the AP-1 signal transduction pathway,VEGF is the key factor of angiogenesis,thereby to restrain angiogenesis.

Objective To evaluate the outcome of living donor liver transplantation(LDLT)for hepatocellular carcinoma(HCC).Methods We retrospectively analyzed the clinical data of 180 patients,who had received LDLT(n=34)or deceased donor liver transplantation(DDLT,n=146)for HCC,compared overall and recurrence-free survival between LDLT and DDLT,and identified the risk factors of tumor recurrence and prognosis by univariate and multivariate analysis.Results The 5-year overall survival and recurrence-free survival rate were 53 % and 58 %,respectively,in DDLT group,and 60 % and 60 %,respectively,in LDLT group.There was no significant difference in overall (P=0.85)and recurrence-free(P=0.89)survival between these two groups.The tumor recurrence rate was 26.5 % in LDLT group,and 17.8 % in DDLT group,respectively(P=0.25).Multivariate COX regression model analysis identified vascular invasion(relative risk 2.118,95 % confidential interval 1.201-4.353,P=0.032)and tumor beyond UCSF criteria(relative risk 3.490,95 % confidential interval 1.862-8.207,P=0.015)as independent risk factors of tumor recurrence,and tumor beyond UCSF criteria(relative risk 8.573,95 % confidential interval 3.016-18.261,P=0.006)as independent predictors of prognosis.Conclusion LDLT is a safe and effective procedure for patients with HCC,but further studies are required for selection criteria of recipients and higher HCC recurrence rate after LDLT.

Objective To provide the practical experience of association of liver partition and portal vein ligation for staged hepatectomy(ALPPS) procedure in portal vein tumor thrombosis(PVT) cases,and to explore its value in PVTT therapy.Methods Three cases of ALPPS were applied to PVTT in Department of Hepatobiliary Surgery of PLA General Hospital from 2015 to 2016.The patients data were retrieved and analyzed retrospectively,including the basic information,preoperative PVTT classification,preoperative Child-Pugh classification,ICG test results,future liver remnant (FLR),FLR growth rate between 2 phase operation,operation time,bleeding volume,postoperative complications,postoperative survival etc.We discussed the detail technology and discuss the surgical procedure combine our experience and the published papers.Results ALPPS was performed successfully in all 3 patients.According to the Cheng's Classification of PVTT,they were classified as type Ⅱ,1 case and Ⅲ type,2 cases.Preoperative liver function was Child-Pugh A grade,average ICG R15 was 7.3％ (4.2％-11.0％),and average FLR was 387 ml (333-484 ml).The mean time interval between 2 phases surgery was 24.7 days (9-50 days) and the average FLR growth rate was 50.3％ (24.4％-82.3％).Morbidity of Clavien-Dindo Ⅲ or more was recorded in 1 case,but no mortality occurred.During follow-up period,2 patients were treated with TACE for tumor recurrence.All patients survived with acceptable life quality till now.The portal vein tumor thrombosis necrosis was observed in all 3 specimens.Conclusions ALPPS is a valuable surgery for effective control of tumor thrombus and radical resection rate in well selected PVVT type Ⅱ and type Ⅲ patients.It is expected to improve the therapeutic effect in combination with TACE and other treatment methods.

Objective To analyze the clinical characteristics and prognosis of patients with thalamic infarction.Methods The clinical data of 47 patients with simple thalamic infarction hospitalized in Beijing Shuili Hospital from January 2016 to January 2022 were retrospectively analyzed.Cranial magnetic resonance imaging and diffusion weighted imaging were used to determine the infarct area of the thalamus,and the clinical characteristics and prognosis of patients with different thalamic infarction regions were observed.Results Among 47 patients with simple thalamic infarction,there were 4 cases of anterior nucleus infarction,22 cases of lateral nucleus infarction,13 cases of medial nucleus infarction and 8 cases of posterior nucleus infarction.The most common clinical manifestations were contralateral hemiplegia,hemidysesthesia and memory impairment.The clinical prognosis of patients with lateral nucleus,anterior nucleus and posterior nucleus infarcts was better,and the prognosis of patients with medial nucleus infarcts was worst.At 90 days of follow-up,the scores of the modified Barthel index were significantly higher than those at admission(P<0.05).Conclusion Patients with thalamic infarction were prone to clinical manifestations such as hemiplegia,hemidysesthesia and memory impairment,but the overall prognosis is good.