Objective To study the inhibition effects of various gastrin-shNAs on gastrin expression in gastric cancer cell line BGC-823. Methods Four nucleotide sequences of shRNA were designed corresponding to various sites of gastrin gene.Four shRNAs were synthesized by in vitro transcription and transfected into gastric cancer cell line BGC-823 at the final concentration of 10nmol/L,20nmol/L,40nmol/L and 80nmol/L respectively.In situ hybridization and immunohistochemistry techniques were applied to investigate the inhibition of gastrin expression and screen the most effective shRNA.The inhibitory effect on gastrin mRNA of screened shRNA was further identified by RT-PCR.MTT assay was used to determine the inhibitory effect of 4 shRNAs at various final concentrations on the growth of BGC-823 cells. Results The gastrin mRNA and protein exression were suppressed distinctly 24,48,and 72hours after transfection,and exhibited time-and concentration-dependent tendency.The highest suppression efficiency on both mRNA(54.27?0.042)% and protein(41.69?0.038)% level occurred 72 hours later in the cells transfected with shRNAs.The RT-PCR result showed that the inhibitory ratio of shRNA3 on gastrin mRNA of BGC-823 was 48.1%.MTT displayed a proliferative inhibition of the BGC-823 cells after transfection of shRNAs with a concentration-denpendent tendency except the shRNA4 treated cells.Conclusion Four gastrin-shRNAs showed a significant inhibition effect on gastrin expression of gastric cancer cell BGC-823 on mRNA and protein level.shRNAs might be the most effective gastrin-shRNA.Inhibited gastrin expression by shRNAs resulted in a significant decrease of proliferative ability of BGC-823 cells.

According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

Objective To study the clinicopathologic features of brain tumors in patients with medically intractable epilepsy. Methods The clinical, radiologic and pathologic features of brain tumors in thirty-six patients with intractable epilepsy encountered during the period from 2008 to 2014 in the Epilepsy Center of Haidian Hospital were retrospectively reviewed. Results There were 18 males and 18 females in thirty-six patients. The mean age of seizure onset and disease duration were (14.05 ± 1.67) years and (10.04 ± 1.19) years respectively. The histological types of brain tumors included ganglioglioma (12/36, WHO gradeⅠ,1/36, WHO gradeⅡ), dysembryeplastic neuroepithelial tumor (2/36, WHO gradeⅠ), pleomorphic xanthoastrocytoma (1/36, WHO gradeⅡ), angiocentric glioma (1/36, WHO gradeⅠ), astrocytoma (4/36, WHO gradeⅡ), oligoastrocytoma (1/36, WHO gradeⅡ, 2/36, WHO gradeⅠ-Ⅱ), oligodendroglioma (1/36, WHO gradeⅠ-Ⅱ,1/36, WHO grade Ⅱ), cavernous hemangioma (4/36) and Sturge-Weber syndrome (1/36). Most of these tumors were located in temporal lobe (25/36, 69.4%). Patients were followed up for 0.5-7 years after operation. One patient was lost for follow up. Seizure outcome after the epilepsy operation revealed that 28 patients (77.8%) had Engel gradeⅠ, 4 patients (11.1%) had Engel gradeⅡ,2 patients (5.6%) had Engel gradeⅢ,1 patient (2.8%) had Engel gradeⅣ. Conclusion Brain tumors in patients with medically intractable epilepsy are almost low grade tumors of the nervous system. Focal cortical dysplasia is existed in most brain tissues from the epilepsy operation. Low grade tumors of the nervous system have close relation with focal cortical dysplasia in patients with medically intractable epilepsy. It is possible that the classifications of pathology diagnosis has connection with prognosis.

Objective:To evalte a novel laparoscopic splenic artery ligation plus devascularization (LSALD) vs. laparoscopic splenectomy and devascularization (LSD) for the treatment of portal hypertention. Methods:From Jan 2014 to Dec 2019, 50 patients undergoing LSALD and 30 patients receiving LSD . We compared the safety and feasibility between LSALD and LSD groups by analyzing the patients′ blood routine, liver function before and after operation, intraoperative condition, postoperative recovery and prognosis.Results:The operation time[(181±72)min vs.(284±72)min , t=-6.205, P<0.01], intraoperative blood loss[(100±50)ml vs.( 700±86 ml), t=-5.166, P<0.01]and blood transfusion rate (28% vs.67%, χ 2=11.471, P<0.01)in LSALD group were significantly more favorite than those in LSD group ( P<0.05). The postoperative exhaust in the LSALD group was earlier than that in the LSD group (2 d vs.3 d, Z=2.361, P<0.05) though the WBC and blood platelet count was higher in LSD group ( P<0.05). Portal vein thrombosis occurred in 10 cases in LSD group and 6 cases in LSALD group (χ 2=5.757, P<0.05). Conclusion:Compared with laparoscopic splenectomy combined with periesophagogastric devascularization, laparoscopic splenic artery ligation combined with periesophagogastric devascularization is less traumatic, helping quick recovery and lower rate of post-op portal vein thrombosis.

This research firstly reported the molecular analysis of ECHO25 (Entric Cytopathic Human Orphanviruses Type 25). To clarify molecular characteristics of the ECHO25 virus isolates in Henan Province and its relationship with the rest of world's isolates,the complete VP1 sequences of the 4 isolates in Henan were successfully amplified by RT-PCR and were compared with other ECHO25 isolates available from GenBank. Compared with the prototype strain JV-4, the nucleotide sequence identity was 79.2%-80.1%, and the amino sequence identity was 89.0%-92.4, the nucleotide sequence identity among the 4 strains isolates in Henan Province was 93.0%-99.0%, the amino sequence identity was 92.4%-97.5%. HN-01 and HN-26 strains had the highest level of homology, the nucleotide homology was 99.0%; All the 4 strains belonged to the B1 genotype.
China ; epidemiology ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; epidemiology ; virology ; Humans ; Molecular Sequence Data ; Phylogeny

Objective To investigate the intelligence standard for diagnose the sub-cretin children and children with mental retardation of socio-cultural type.Methods The full intelligence quotient(IQ),verbal intelligence quotient(VIQ)and performance intelligence quotient(PIQ)was tested by Wechsler scale(C-WISC)for mild iodine deficiency disordem children,children living in abnormal socio-cultural condition and normal children aged 7～14 years old in Qinba mountain area.The test results had been compared between the groups.Results There were no significant difference between psychomotor functioning well children and children living normal sociocuhural condition in VIQ,PIQ and full IQ(89.24±18.44 vs 90.75±17.58,87.58±15.78 vs 88.95±15.56,87.42±17.84 vs 89.02±17.18,t=1.14,1.19 and 1.24,respectively,all P＞O.05).PIQ and full IQ were significantly lower in mild iodine deficiency disorders children than in children with abnormal socio-cultural background (65.81±10.22 vs 72.33±13.23,62.42±12.31 vs 68.13±14.54,t=3.26,2.55,P＜0.01 or＜0.05,respectively).But the VIQ was not significantly different between these two groups.The average difference of VIQ and PIQ among mild iodine deficiency disorders children wag-0.32 without significant difierence(t=0.28,P＞0.05),however it was-2.91 among children under abnormal socio-cultural condition with significant difierenee(t=-3.59,P＜0.01).Conclusions IQ for iodine deficiency disorders children is characterized by that VIQ is damaged in parallel with PIQ,while that in children under abnormal soeio-cuhural condition is marked by that VIQ is retarded more severely than PIQ,which ean be used as an intelligence standard for differentiating the sub-cretin children from children wjth socio-cuhural mental retardation.

strains (isolated in 2008). Conclusion The HENAN08 strain might belong to the same genogroup with AnhuiFY08 and Zhejiang08 strains as C4 gene subtypes.

Objective:To observe the correlation between the frontal P-wave axis and the severity of chronic obstructive pulmonary disease (COPD) and the prognosis evaluation system.Methods:Patients with COPD>45 years old who were followed up in the outpatient department of Hunan Chest Hospital from January to July 2022 were continuously selected as subjects. At the same time, the healthy people who examined in the health management center of our hospital were in the control group. Both groups of subjects completed electrocardiogram and pulmonary function tests. The level of frontal P-wave axis and the results of pulmonary function examination were recorded, and the differences of frontal P-wave axis between the COPD group and the control group were compared, so as to clarify the value of frontal P-wave axis in the diagnosis, disease severity and prognosis evaluation of COPD.Results:The level of forced expiratory volume in the first second/forced vital capacity(FEV1/FVC )in the COPD group was significantly lower than that in the control group, while the level of P-wave axis was significantly higher than that in the control group (all P<0.05). The receiver operating characteristic (ROC) curve of P-wave axis showed that the AUC of P-wave axis in predicting COPD was 0.96 ( P<0.001), the best cut-off value was 63.80, the sensitivity was 0.89, and the specificity was 0.93. There were significant difference in P-wave axis level, the forced expiratory volume in one second to forced vital capacity ratio (FEV 1%pred), body mass index (BMI) and BMI, airflow obstruction, dyspnea, and exercise capacity (BODE) index between groups according to the degree of airflow limitation (all P<0.05). Correlation analysis showed that P-wave axis level was positively correlated with BODE index ( r=0.77, P<0.001), and negatively correlated with pulmonary function FEV 1%pred ( r=-0.76, P<0.001). Conclusions:There is a good correlation between the level of frontal P-wave axis and the severity of COPD and the prognosis evaluation system, which has clinical application value.

OBJECTIVETo study the effect of total flaveos of Gymostemma pentaphyllum on the protein expression of apoptosis-associated Fas/FasL gene and tumor necrosis factor-alpha (TNF-alpha) concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation (H/R).

METHODA cultured primary neonatal rat cardiomyocytes model with H/R was erected, experiments were divided into six groups, (1)control group, (2)H/R group, (3)15 mg x L(-1) TFG plus H/R group, (4)45 mg x L(-1) TFG plus H/R group, (5) 105 mg x L(-1) TFG plus H/R group, (6)105 mg x L(-1) TFG group. TNF-aconcentration in cultured cardiomyocytes with H/R, was determined by ELISA method, the protein expression of Fas/FasL genes were estimated by immunohisto-chemistry.

RESULTAfter cardiomyocytes were made with H/R, Compared with control group, the positive expression index (PEI) of Fas/FasL proteins in cardiomyocytes increased significantly, Compared with H/R groups, the PEI of Fas/FasL proteins were lower significantly in groups with different dosages of TFG (P < 0.05). TFG inhibited the secretion of TNF-alpha from myocardial cells and increased the survival rate of myocardial cells.

CONCLUSIONThe protein expression of apoptosis-associated Fas/FasL genes increased during H/R. The TFG can protect myocardium against H/R injury by decreasing the production of TNF-alpha, downregulating the protein expression of Fas/FasL genes, and then inhibiting myocyte apoptosis.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Fas Ligand Protein ; metabolism ; Flavones ; isolation & purification ; pharmacology ; Gynostemma ; chemistry ; Immunohistochemistry ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Oxygen Consumption ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo analyze the prevalence trend, prevalence characteristics and influence factors of Japanese B Encephalitis (JE) in Henan province.

METHODSThe data that of 64 401 JE patients in Henan from 1980 to 2008 were statistically analyzed by SPSS12.0 and EXCEL2003 software. Luoshan, Xinan, Xihua, Deng county and Hua county were chosen as monitoring sites. The mosquito specimens were gathered with the artificial hour method and the mosquito curtain method, the mosquito density was calculated one time each ten day period from May to July. At the same time, 30-40 newborn pig blood samples were gathered each ten-day period and the pig serum JE IgG antibody was detected by ELISA method.

RESULTSThe Cumulative incidence of JE was 64 401 cases in Henan province from 1998 to 2008, the range of incidence rate was from 0.34/100 000 (315/93 599 969) to 6.72/100 000 (5246/78 076 567); The average incidence of JE was 4.39/100 000 (3530/80 381 469) from 1980 to 1994; The average incidence of JE was 0.86/100 000 (811/94 217 549) from 1995 to 2008; In 2008, the incidence rate reached the lowest point for 0.34/100 000 (315/93 599 969); The incidence occurred mainly in July-September, accounting for 89.40% of the total cases (57 572/64 401); the patients were concentrated mainly in 5 cities, which were Xinyang, Nanyang, Zhumadian, Zhoukou, Luoyang, accounting for 81.02% (52 175/64 401). The 0 - 14 years old age group was the dominant group (79.01%, 50 884/64 401). In Luoyang city, incidence of >/= 15 years old group was significantly increased (57.83%, 2120/3666), the constitution of JE incidence were significantly different between 0 - 14 years old group and >/= 15 years old age group (chi(2) = 2705.32, P < 0.05) in Henan province and Luoyang city. The different density of the mosquitoes and the different positive-times for 50% of the antibodies of JE in piglets on the monitor sites showed the intensity of JE disease.

CONCLUSIONThe incidence of JE showed a decreasing trend, seasonal, regional characteristics and age distribution difference in Henan province. The monitoring of host animal pig JE antibody level and the medium mosquito density may forecast the JE prevalence tendency. To control the incidence in the younger groups in Henan province, older age group in Luoyang city and high-incidence areas, it is important to strengthen the monitoring and forecasting measures to prevent JE in Henan province.

Adolescent ; Animals ; Child ; Child, Preschool ; China ; epidemiology ; Culicidae ; Encephalitis, Japanese ; epidemiology ; prevention & control ; Environmental Monitoring ; Epidemiological Monitoring ; Humans ; Incidence ; Infant ; Population Surveillance ; Swine ; Universal Precautions