Objective To construct the stably over-expressing hB7-H3 gene human pancreatic cancer cell line PANC1,and provide tools for further investigating the function of hB7-H3.Methods hB7-H3 gene fragment was inserted into lentiviral plasmid GV287 carrying GFP to construct recombinant hB7-H3-GV287 plasmid vector.293T cells were transfected,and the GFP expression was evaluated under fluorescence microscopy.Western Blot wasused to detect the expression of hB7-H3 protein.Lentiviral vectors were packaged and the titer was determined.The recombinant hB7-H3 expressed lentivirus was used to infect PANC1 cell.Flow cytometry was applied to detecte the percentage of GFP and hB7-H3 positive cells.Real-time PCR and Western Blot was used to verify the mRNA and protein expression of hB7-H3.Self-cyclizing GV287 plasmid served as negative control (NC).Results PCR amplified fragment of recombinant plasmid was around 1 368 bp,and no amplified production of NC plasmid was observed.The DNA sequencing of recombinant plasmid was completely consistent with the designed fragment,indicating that hB7 H3-GV287 plasmid was successfully constructed.293T cells transfected with recombinant plasmid expressed hB7-H3 protein,while those cells transfected with NC plasmid did not express hB7-H3 protein.The virus titer of lentiviral packaged recombinant hB7-H3 plasmid was 2 × 108 TU/ml.The percentage of hB7-H3 positive cells,hB7-H3 mRNA and protein expression in PANC1 cells infected with cells infected with hB7-H3 lentivirus was 94.3％,5.09 ± 0.24 and 2.85 ± 0.27,respectively,which was obviously higher than 18.5％,1.28 ± 0.53 and 0.44 ± 0.69 in cells infected with NC lentivirus,and the differences were statistically significant (P value ＜0.01).Conclusions A human pancreatic cancer cell line PANC1 stably over-expressing hB7-H3 was successfully constructed.

Objective To discuss the influence of intravenous anesthesia with procaine on the release of central nervous Ach. Methods 50 samples of plasma were collected from healthy people, and 30 samples of plasma from cases prior to surgical operation, during anesthesia and after operation respectively. The activity of Ach in the plasma was measured. Then observe the dropping speed and concentration of procaine at different time during anesthesia, and the subsequent changes of the activity of plasma Ach and the slowdown of its activity. Results The data showed that the activity of plasma Ach prior to operation was considerably lower than that of the control group (P

Objective To compare the performance of two commercially available ELISA kits for detection of diphtheria IgG antibody to provide data for the proper kit choice. Methods Thirty-three serum samples from staff of the capital airdrome and two standard sera were tested for the evaluation of the kits.Commercial ELISAs were compared with respect to their reproducibility(intra-assay variation and inter-assay variation)and accuracy.Results For the detection of standard serum,intra-assay variation(Coefficient of Variation,CV)was 13.5507% for IBL kit and 5.9541% for Virion kit,the inter-assay variation(CV)was 9.9890% for IBL and 2.4728% for Virion kit.For the serum sample,intra-assay variation(CV)was 14.5270% for IBL and 11.0612% for Virion,the inter-assay variation(CV)11.0039% for IBL and 8.1506% for Virion kit.For the detection of standard serum,inter-assay variation(CV)was 9.9890% for IBL and 2.4728% for Virion.The accuracy was analyzed by variation between the actual concentration and measured concentration of standard serum,the CV was 17.6889% for IBL and 1.3966% for Virion.Conclusion The intra-assay and inter-assay variation of the two kits are generally acceptable(CV

Objective To observe the inhibition effect of selective cyclooxygenase-2 inhibitor (celecoxib) on the experimental choroidal neovascularization (CNV).Methods Thirty 8-10 weeks old healthy male Brown-Norway(BN) rats were randomly divided into the control,laser and celecoxib group,with 10 rats in each group.At the dosage of 50 mg/kg,celecoxib was garaged twice per day.After 7 days,experimental CNV was induced by Krypon laser on laser group and celecoxib group.Fundus fluorescein angiography (FFA) was performed on days 3,7,14,21,30 after laser photocoagulation.On days 21 after photocoagulation,5 rats in each group were sacrificed and the relative thickness of CNV membranes,the expression of COX-2,vascular endothelial growth factor(VEGF) and matrix metalloproteinase-2 (MMP-2) Were studied by histopathologic or immunohistochemistry examination.Results On days 21 after photocoagulation,the incidence of CNV in the celecoxib group is significantly lower than that in the laser group (X~2=7.1068,P=0v0077);the relative thickness of the CNV membranes in the celecoxib group is reduced 41.38% compared to the laser group,the difference is statistically significant (t=16.7600,P=0.0000).COX-2,VEGF and MMP-2 expression in the CNV membrane of celecoxib group were significantly lower than in control group (t=5.7100,5.8400,8.0200;P=0.0000);the COX-2,VEGF and MMP-2 expressions in choroid and retina of control group were weak.Conclusion Prophylactic celecoxib can reduce the expression of VEGF and MMP-2 by inhibiting COX-2,and prevent the CNV induced by laser photocoagulation.

Objective To evaluate the effect of azithromycin on mycoplasmal pneumonia (MP).Methods We divided 90 MP cases into azithromycin and erythromycin treatment groups. In azithromycinThe pyretolysis time, cough improvement time, the disappearing time and the mean length of hospitalization of azithromycin group were shorter than that of erythrornycin group. The local ache, stomach and intestinal tract adverse reaction, and damage of hepar function were less than these in erythromycin group. ConclusionAzithromycin is an effective and safe drug to MP.

Objective To observe the clinical effects of different doses of Xuebijing injection in the treatment of sepsis. Methods Eighty patients with sepsis from First Center Clinical College of Tianjin Medical University were selected and randomly divided into four groups:the control group,Xuebijing small,middle and large dose groups(each, 20 patients). All patients in the four groups were given Western medicine conventional treatment,whereas in the Xuebijing small,middle and large dose groups,additionally they were intravenously dripped with Xuebijing injection 50 mL, 2 times a day,100 mL,2 times a day and 100 mL,4 times daily respectively. The patients' vital signs,blood routine, interleukin-18(IL-18),procalcitonin(PCT)and C- reactive protein(CRP)were detected before treatment(0 d)and 3,7 and 14 days after Xuebijing injection,28-day and 60-day death rate were observed. Results With prolonged treatment,the body temperature,heart rate(HR),respiratory rate(RR),white blood cell count(WBC),IL-18,PCT, CRP levels were all decreased in each group(all P<0.05),and on 14 days they reached the valley;when compared with middle,small dose Xuebijing groups and control group,in the Xuebijing large dose group they were decreased the most significantly over the same period〔temperature (℃):36.22±0.55 vs. 36.55±0.99, 36.87±1.05, 36.93±1.07,HR(beats/min):82.11±8.99 vs. 93.45±10.02,109.02±11.44,113.13±10.34,RR(breaths/min):15.48±6.45 vs. 17.33±6.94,20.35±7.19,22.34±6.67,WBC(×109/L):5.35±1.37 vs. 8.35±2.98,10.54±3.22, 13.33±4.78,IL-18(ng/L):107.36±55.89 vs. 136.55±80.19,177.78±102.32,193.93±98.07,PCT(μg/L):0.33±0.19 vs. 0.66±0.22,1.62±0.44,2.78±0.34,CRP(mg/L):15.40±1.37 vs. 48.34±8.98,100.45±30.67, 131.92±49.53〕,the differences all being statistically significant(all P<0.05). The 28-day and 60-day death rate in Xuebijing large dose group were significantly lower than those in control group,Xuebijing small,middle dose groups (28-day:25% vs. 40%,40%,45%;60-day:30% vs. 45%,40%,50%). Conclusion The time for onset of therapeutic effect in large dose Xuebijing group is short,and reduce the levels of inflammatory mediators significantly, and improve the prognosis of patients with sepsis,therefore it can be used clinically.

Objective To improve the diagnosis and treatment of von H ippel-Lindau (VHL) disease.Methods The clinic data of one kindred with VHL disease type Ⅱ B was reviewed including clinical manifestation,imaging,pathology and therapy.A 40-year-old male complained of the left upper extremity numbness for 6 months,and a variety of visceral tumors were found 3 months ago.Contrast-enhanced MR imaging showed multiple brain tumors.CT showed left kidney tumor,pancreatic tumor,pars-aortic tumor and left adrenal tumor.Fundoscopy showed multiple retinal hemangioblastoma.PET-CT discovered abdominal multiple tumors.And pedigree analysis was to determine the family medical history,and 6 members got genetic screening.Results In the kindred with VHL disease type Ⅱ B,9 members (30％) out of 40members in 4 generations got the disease.Six members got genetic screening,and the result showed 5 (5/6) members had mutation.Three (3/5) members with 3 sites genetic mutation showed clinical manifestation,1 (1/5) members with 3 sites genetic mutation without clinical manifestation found brain tumors by MRI,and 1 ( 1/5 ) member with 1 site genetic mutation did not find disease by comprehensive checkup.The main mutation located at exon 1 in chromosome 3p25 of VHL gene.All mutation was hetcrozygous mutation.The 295,337 and 337 nucleotide thymine of the VHL gene were substituted by cytosine,cytosine and adenine,which made the 98th,112th and 112th tyrosine substituted by histidine and asparagines.One member with 1 site mutation had 98th tyrosine substituted by cytosine.The first operation was to remove brain tumor,and the second operation was to remove adrenal tumor,para-aortic tumor and renal tumor.Pathology of the brain tumors showed hemangioblastoma,and the retroperitoneal tumors were clear cell carcinoma,paraganglioma and pheochromocytoma.Followed up for 6 months after operation,no relapse occurred.Conclusions VHL disease is a relatively rare autosomal dominant disorder.Comprehensive management of patients should also include genetic counseling and screening for other manifestations of the disease process.Genetic testing might be helpful in early detection of asymptomatic VHL patients.Members having gene mutation should be followed up strictly.Surgical management of VHL disease should be decided on the base of comprehensive assess.Multiple lesions could be cut off in one operation.For patients with pheochromocytoma,pheochromocytoma shuld be handled first.

Objective: To observe the clinical efficacy of combined acupuncture and speech rehabilitation in treating postapoplectic aphasia. Methods: Sixty subjects were randomized into two groups. 30 cases in the treatment group were treated by both speech rehabilitation and acupuncture therapy, while 30 cases in the control group were treated by speech rehabilitation only. After 30-day treatments, the clinical efficacy was observed. Results: The total effective rate and speech score were higher in the treatment group than in the control group (P<0.05). Conclusion: Combined acupuncture and speech rehabilitation is quite effective for postapoplectic aphasia.

Based on the high solubility of azodicarbonamide (ADC) in hot solution and its electrostatic interaction with Nafion film,a new electroanalytical method was developed for the determination of ADC by using Nafion film electrode.The effect of temperature on the solubility of ADC and the mechanism of the reduction reaction of ADC on Nafion film electrode were investigated.Under the experimental conditions such as water bath at a constant temperature of 80 ℃,pH 6.0 and optimal test parameters,the differential pulse voltammetric response was proportional to the concentration of ADC in the range of 0.93-10.5 μg/L,and the detection limit was estimated to 0.58 μg/L.The relative standard deviation was less than 5.86 ％ and the recovery was 95.8％-104.0％ for the determination of the ADC in flour samples.The semicarbazide and nitrofurazone did not interfere with the determination of ADC.

Recently,the new term lamellar hole-associated epiretinal proliferation was introduced to describe a specific epiretinal proliferation.Different from conventional epimacular retinal membrane,lamellar hole-associated epiretinal proliferation (LHEP) can be found around the edge of lamellar macular holes and part of full thickness macular holes.It is defined as a thick homogenous layer of yellowish material without any contractive properties on the epiretinal surface with medium reflectivity on optical coherence tomography images.Immunocytochemical analysis showed the presence of glial cells,fibroblasts,hyalocytes and collagen type Ⅱ.Electron microscopy revealed fibroblasts and hyalocytes as predominant cell types,densely packed in cell agglomerations.LHEP is a secondary event in lamellar macular hole formation and may represent a repair process after large and deep retinal defect.Further studies on its clinical features and clinical significance are still required.