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 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.

Past decade,plants as production systems for recombinant proteins has been actively investigated,and is one of the most active research fields in life sciences. Plants are attractive as protein factories because they with advantages of cost,scale,and safety,and emerge capability to produce pharmaceutical production with bioactivity proved by previously studies. The plant-made pharmaceuticals possess enormous viability for market and wide prospects for application. Based on those reasons,the rise of plant bioreactor was reviewed,and research advance on the expression systems and plant-made recombinant proteins was introduced. Bottleneck problems and adopted strategies using plant system to produce recombinant proteins and research hotspot in the near future were summarized,and prospect of plant bioreactor in future was described. Synchronously,some opinions including research progress of plant bioreactor,differences to foreign and strategies in the future were discussed.

Sleep plays an important role in the modulation of endocrine function and energy metabolism;hormone levels are markedly different during a sleepless night compared with a night of normal sleep.Sleep deprivation may change the levels of cortisol,growth hormone(GH)and thyrotropin(TSH)at night.In addition,sleep curtailment disturbs the balance of anorexi- genic(leptin)and orexigenic(ghrelin)factors,and does harmful to glucose tolerance.

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 assess the factors affecting survival of malignant pheochromocytoma/malignant paraganglioma.Methods The clinical data of 85 malignant pheochromocytoma/malignant paraganglioma patients were analyzed retrospectively.All the patients were followed up,the duration from 2 months to 283 months,the median time was 48 months.The overall survivals of 5 years and 10 years were calculated.The possible affecting factors,such as gender,age,tumor function,metastatic sites,metastatic fields,incipience or recurrence,the time to recur,were analyzed.Survival differences between groups were compared with the log rank test.Multivariate analysis was performed by using Cox regression analysis to detect variables independently associated with survival.All P values were 2 tailed with P ＜ 0.05 considered statistically significant.All statistical analysis was done by SPSS 17.0 software package.Results The overall median survival time was 62 months,5-year survival rate was 44.7％,10-year survival rate was 11.8％.Gender (P =0.649),age (P =0.228),incipience or recurrence (P =0.217) had no significant effect on the survival time.Significance was found in tumor function (P =0.034),metastatic sites,metastatic fields (P =0.009),the time to recur (P =0.003).The median survival time of patients with nonfunctioning tumor and functioning tumor was 90 months and 37 months.The median survival time of patients with multiple system metastases and single system metastasis was 37 months and 117 months.For the patients with single system invaded,there were 15 cases (26.8％) of bone metastasis,10 cases (17.9％) of lymph node metastasis,8 (14.3％) of liver metastasis,7 (12.5％) of lung metastasis,6 (10.7％) of renal metastasis,6 (10.7％) of intestine metastasis and 4 (7.1％) of brain metastasis.The median survival time was 110,77,28,14,26,8 and 19 months.The median survival time of patients with lung,liver,intestine metastasis were shorter than the other patients (P ＜ 0.05).Fifty-five patients were found recurrence after operation,recurrent time was from 4 to 65 months,and the median time was 22 months.The survival time of patients with recurrence within 2 years was shorter than patients above 2 years (P =0.003).Conclusions The prognosis of malignant pheochromocytoma/malignant paraganglioma is poor.Gender,age,incipience or recurrence could not affect the overall survival time.The survival time has relationship with the function of tumor,metastatic field,metastatic sites and time to recurrence.The patients with nonfunctioning tumor have longer survival time than those with functioning tumors.The survival time of patients with single system invaded is longer than ones with multiple systems invaded.The common metastatic sites are bone,lymph node,liver,lung,kidney,intestine and brain.Patients with intestine metastasis have worst prognosis.The patients with only bone and lymph node metastasis have best prognosis.Patients with recurrence above 2 years have a better prognosis than those within 2 years.

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.

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.

Objective To investigate the effect of has-miR-663 on proliferation and apoptosis of human adrenal cortical cell line SW-13.Methods Based on the first stage study,in adrenocorticotropinindependent macronodular adrenal hyperplasia (AIMAH),hsa-miR-663 was up-regulated.The synthetic hsa-miR-663 mimics and inhibitors were transfected the SW-13 cell line.Real-time quantitative PCR (RT-PCR) tested the effect of the transfection.According to the results of RT-PCR,cell line was divided into five groups,including blank group,mimics NC group,inhibitor NC group,mimics group,inhibitor group.Then proliferating of the cell was tested by MTT and the apoptosis by Annexin V-FITC/PI.Result By RT-PCR,mimics group showed that has-miR-663 was over-expressed,and inhibitor group were knocked down.MTTshowed that,compared with NC group and Blank group,SW-13 cell line proliferated faster in mimics group and slower in inhibitor group.Annexin V-FITC/PI showed that apoptosis of SW-13 cell line was not different in all the five groups.Conclusion Up-regulating has-miR-663 could accelerate proliferation of adrenal cortical cells.The microRNA might play some role in the pathogenesis of AIMAH.