Objective To investigate the protective effect of oxygen-glucose deprivation preconditioning(OGDPC)on PC 12 cells damaged with oxygen-glucose deprivation/reperfusion(OGD/R)and its underlying mechanism.Methods PC 12 cells were randomly divided into Control,OGDPC,OGD/R and OGDPC+OGD/R groups.The morphology and quantity of mitochondria were observed by transmission electron microscopy(TEM)and Mito-tracker staining.The expression of OGDH,PDHA1,COX4 and HSP60 was detected by Western blotting.Mitochondrial membrane potential and ATP production were evaluated by JC-1 and ATP assay kit.TUNEL staining was used to assess cell apoptosis,and translocation of AIF and EnodoG was observed by immunofluorescence staining and TUNEL staining.Results Compared with the OGD/R group,the mitochondrial ultrastructure damage were alleviated,the expression levels of OGDH,PDHA1 and COX4 were enhanced(P＜0.05),that of HSP60 was decreased(P＜0.05),the mitochondrial membrane potential and ATP level were increased(P＜0.05),the cell apoptotic rate was reduced(P＜0.05),and AIF and EndoG were less aggregated to the TUNEL-positive nucleus in OGDPC+OGD/R group(P＜0.05).Conclusion OGDPC can alleviate the injury of PC12 cells caused by OGD/R,and the potential mechanism may be related to improving mitochondrial structure and function,and reducing cell apoptosis via mitochondrial pathway.

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.

Objective To explore the clinical value of oral ultrasonic contrast agent ultrasonography (OUCAUS) combined with serum monocyte chemoattractant protein-1 (MCP-1) and cell adhesion molecule-1 (CAM-1) measurement in preoperative staging of stomach carcinoma.Methods 800 gastric cancer patients were diagnosed by electric gastroscopy and OUCAUS.The preoperative staging was measured by OUCAUS and compared with pathologic staging,and serum levels of MCP-1 and CAM-1 were measured with ELISA.Results The total accuracy rate of OUCAUS was 79.9％ in estimating invasive depth of stomach neoplasm,82.9％ in estimating lymphatic metastasis and 88.6％ in estimating distant metastasis respectively.The expression levels of MCP-1 and CAM-1 in serum were closely correlated with invasive degree,lymphatic metastasis,distant metastasis and pathologic staging (all P ＜ 0.05).The total accuracy rate of combining OUCAUS and MCP-1,CAM-1 was 93.0 ％ in estimating invasive depth,93.9％ in estimating lymphatic metastasis and 98.6％ in estimating distant metastasis respectively.The total accuracy rate of combining OUCAUS and MCP-1,CAM-1 in estimating invasive depth,lymphatic metastasis and distant metastasis was significantly higher than that of by OUCAUS alone.Conclusions MCP-1 and CAM-1 serum levels are closely correlated to pathologic staging of gastric cancer.Combining OUCAUS and MCP-1,CAM-1 can increase the accuracy rate determining invasion and metastasis in gastric cancer.

OBJECTIVETo establish a modified method for microculturing whole human blood for cytogenetic analysis.

METHODSA novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.

RESULTSThe SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.

CONCLUSIONA modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.

Adolescent ; Adult ; Cell Culture Techniques ; methods ; Cytogenetics ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Mitotic Index

OBJECTIVE: To investigate the correlation between Helicobacter pylori (H. Pylori) infection and polymorphism of adiponectin gene promoter -11391G/A, extracellular superoxide dismutase (EC-SOD) gene in nonalcoholic fatty liver disease (NAFLD).
 METHODS: From June, 2010 to July, 2014, a hospital-based 1:1 matched case-control study was carried out, with 600 cases of NAFLD and 600 healthy people in the First Affiliated Hospital of Xinxiang Medical University. The genetic polymorphisms of adiponectin gene promoter -11391G/A and EC-SOD were analyzed by polymorphism-polymerase chain reaction (PCR) technique in peripheral blood leukocytes of the subjects. 14C-urea breath test (14C-UBT) was used to test 14C disntegration per minute (DPM) for evaluating the infections status of H. Pylori. The synergistic effect between the two mutants and the gene-environment interaction of the genotypes with H. Pylori infection were analyzed.
 RESULTS: The frequencies of -11391G/A (AA) and EC-SOD (CG+GG) were 50.67% and 50.33% in NAFLD cases, 23.83% and 24.17% in healthy controls, respectively. Statistical tests showed significantly higher frequencies of -11391G/A (AA) and EC-SOD (CG+GG) in the NAFLD group (-11391G/A: P=0.0051; EC-SOD: P=0.0057). The risk of NAFLD with -11391G/A (AA) was significantly higher than those with -11391G/A(GG+GA) (OR=3.2822, 95% CI 1.9170 to 5.2039). The individuals who carried EC-SOD (CG+GG) had a high risk of NAFLD (OR=3.1800, 95% CI 1.7974 to 5.2391). Combined analysis of the polymorphisms showed that percentage of -11391G/A (AA)/EC-SOD (CG+GG) in the NAFLD group was significantly higher than that in the control groups (25.50% vs 5.83%, P=0.0039). The people who carried with -11391G/A (AA)/EC-SOD (CG+GG) had a high risk of NAFLD (OR=10.3190, 95% CI 8.1869 to 20.5102). The H. Pylori infection rate in the NAFLD group was significantly higher than that in the control group (OR=3.1667, 95% CI 1.9139 to 5.7443, P=0.0062), and statistical analysis suggested a positive correlation between H. Pylori infection and NAFLD with -11391G/A (AA) and EC-SOD (CG+GG) (-11391G/A: γ=1.8532; EC-SOD: γ=1.7899).
 CONCLUSION These carriers of -11391G/A(AA) and EC-SOD (CG+GG) genotypes may have a high risk of NAFLD, and the gene genotypes can interact with H. Pylori infection in the pathogenesis of NAFLD. Therefore, effective prevention measures for NAFLD should consider eradicating H. Pylori or regulating gene expression.
Adiponectin ; genetics ; Case-Control Studies ; Gene-Environment Interaction ; Genotype ; Helicobacter Infections ; complications ; genetics ; Helicobacter pylori ; Humans ; Non-alcoholic Fatty Liver Disease ; complications ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Risk Factors ; Superoxide Dismutase ; genetics

OBJECTIVE: To investigate the interaction between polymorphism of Toll-like receptor 4 (TLR4) gene G11367C in 3' untranslated region (UTR) and inhibitor of nuclear factor kappaB (IκB)-α 
Hae III in acute pancreatitis (AP) and the degree of severity.
 METHODS: A total of 450 patients with confirmed AP (AP group), who came from the First Affiliated Hospital of Xinxiang Medical College from May 2013 to June 2015, were divided into a mild AP subgroup (MAP subgroup), a moderately severe AP (MSAP subgroup), and a severe acute AP (SAP subgroup) (n=150 in each group). One hundred fifty healthy persons were served as a control group. There was no significant difference in age, gender, ethnicity and birthplace among all groups. The genetic polymorphisms of TLR4 gene G11367C in 3' untranslated region and IκB-α Hae III were analyzed by polymerase chain reaction (PCR). Eligible participants were personally interviewed by a questionnaire. Unconditional logistic regression model and single factor analysis were performed to calculate the adjusted odds ratios (OR) and 95% confidence intervals (95% CI) of G11367C and IκB-α Hae III polymorphisms, respectively. The interaction of nucleotide polymorphisms was analyzed.
 RESULTS: The frequencies of G11367C (GC), IκB-α Hae III (AG) and IκB-α Hae III (GG) were 69.56%, 33.78% and 36.22% in the AP group; 49.33%, 24.67% and 26.00% in the MAP subgroup; 70.67%, 34.67% and 36.67% in the MSAP subgroup; 88.67%, 42.00% and 46.00% in the SAP subgroup and 26.67%, 14.00% and 14.67% in the control group, respectively. There was significant difference in the frequencies betweenc the AP group and the control group, or among each AP subgroup (all P<0.01). The risk of AP was significantly increased in the subjects with G11367C (GC) genotype (ORAP=6.2828, ORMAP=2.6776, ORMSAP=6.6250, ORSAP=21.5147), which was also increased in those with IκB-α Hae III (AG) genotype (ORAP=5.7369, ORMAP=2.5277, ORMSAP=6.1824, ORSAP=17.8572) and in those with IκB-α Hae III (GG) genotype (ORAP=5.8724, ORMAP=2.5902, ORMSAP=6.4027, ORSAP=18.9022). The combined analysis of the polymorphisms showed that the percentage of G11367C (GC)/ IκB-α Hae III (GG) in the AP group, the MAP subgroup, the MSAP subgroup, the SAP subgroup and the control groups was 26.44%, 12.67%, 26.00%, 40.67% and 4.00%, respectively, with significant difference in the frequency among all groups (all P<0.01). The people who carried with Pro12Ala (AA)/Pro198Leu (LL) had a high risk of AP (ORAP=30.1314, ORMAP=6.7612, ORMSAP=39.5000, ORSAP=401.5833), and the statistical analysis suggested a positive interaction between Pro12Ala (AA) and Pro198Leu (LL) in increasing the risk of AP (All γ>1). Similarly, there were also positive interactions in the pathogenesis of AP between G11367C (GC) and IκB-α Hae III (AG) (All γ>1). 
 CONCLUSION These carriers of G11367C(GC), IκB-α Hae III(AG) and IκB-α Hae III (GG) genotypes may have a high risk of AP occurency, and there are significant interactions between genetic polymorphisms of G11367C and IκB-α Hae III, which increaes the risk of the occurrence and development of AP.
3' Untranslated Regions ; Acute Disease ; Deoxyribonucleases, Type II Site-Specific ; Ethnic Groups ; Genetic Predisposition to Disease ; Genotype ; Humans ; I-kappa B Kinase ; Logistic Models ; NF-KappaB Inhibitor alpha ; Odds Ratio ; Pancreatitis ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Toll-Like Receptor 4

Objective To investigate the relationship of the interaction between age and polymorphisms of E-selectin gene A561C, chemokine receptor CCR2 gene A190G with the susceptibility, invasion and metastasis of gastric carcinoma.Methods Based on tumor-node-metastasis (TNM) staging classification, 750 patients with confirmed gastric carcinoma in our hospital from December 2011 to November 2014 were divided into 5 groups: stage Ⅰ, stage Ⅱ , stage Ⅲ, stage Ⅳ and stage 0 (n=150, each).No significant difference was observed in gender, ethnicity, birthplace and living habits among the 5 groups.Meanwhile, 750 healthy controls were selected in this study during the same time, and there was no significant difference in gender, ethnicity and birthplace between the healthy controls and patients with gastric carcinoma.The genetic polymorphisms of E-selectin gene A561C and chemokine receptor CCR2 gene A190G were analyzed by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) in peripheral blood mononuclear cells (PBMs).Results The frequencies of CC (A561C) and GG (A190G) genotypes were 56.5％ and 56.8％ respectively in gastric carcinoma cases and 22.8％ and 23.1％ respectively in healthy controls, with statistically significant differences in the distribution frequencies between the two groups (P＜0.01 for all).The risk for gastric carcinoma significantly increased in subjects with CC (A561C) genotype (OR=4.4038, 95％CI=2.9421-7.2397) and in GG (190A/G) genotype (OR=4.3852, 95％ CI =2.8207-7.4942).Combined analysis of the polymorphisms showed that the distribution frequency of CC (A561C) genotype / GG (190A/G) genotype in gastric carcinoma cases and healthy controls was 46.4％ and 11.9％ respectively (P＜0.01).The positive interactions of age with CC (A561C) genotype and GG (190A/G) genotype for the risk of invasion and metastasis of gastric carcinoma were found (γ＞1 for both).The distribution frequencies of CC (A561C) genotype and GG (190A/G) genotype were 50.0％ and 50.0％ in stage Ⅰ , 63.4％ and 64.0％ in stage Ⅱ ,69.3％ and 69.3％ in stage Ⅲ, 76.7％ and 77.3％ in stage Ⅳ, and 23.3％ and 23.3％ in stage 0 respectively.Statistically significant differences were found in the distribution frequencies between stage 0 and the other 4 stages (P＜0.01 for all).The risks for the invasion and metastasis of gastric carcinoma were significantly increased in subjects with CC (A561C) genotype (ORⅠ-Ⅳ =3.2857-10.7959) and in those with GG (190A/G) genotype (ORⅠ-Ⅳ =3.2857-11.2101).Combined analysis of the polymorphisms showed that distribution frequency of CC (A561C) genotype / GG (190A/G) genotype had significant differences between the stage Ⅰ ～Ⅳ and stage 0 (39.3％, 53.3％, 59.3％,68.0％ vs.12.0％, P＜0.01).The proportion of elderly subjects were higher in Grade Ⅰ ～Ⅳ than in Grade 0 (51.3％, 62.7％, 70.0％, 75.3％ vs.26.7％, P＜0.01 for all).The risk for invasion and metastasis of gastric carcinoma was significantly increased in elderly patients (ORⅠ-Ⅳ =2.9001 ～8.3986).The positive interactions of age with CC (A561C) genotype and GG (190A/G) genotype for the risk of invasion and metastasis of gastric carcinoma were found (γ＞ 1 for All).Conclusions Age and E-selectin gene A561C (CC) and chemokine receptor CCR2 gene A190G (GG) are the risk factors for the invasion and metastasis of gastric carcinoma, and the interactions between age and genetic polymorphisms increase the risk of invasion and metastasis of gastric carcinoma.

Objective To explore the value of the variation of clinical pulmonary infection score (CPIS) and serum procalcitonin (PCT) in diagnosis and treatment evaluation in patients suffering from active pulmonary tuberculosis with complication of ventilator-associated pneumonia (VAP).Methods A retrospective analysis was carried out in 58 VAP patients from June 1,2009 to December 30,2014 in the respiratory intensive care unit.According to the patient suffering from tuberculosis or not,patients were divided into two groups.The PCT and CPIS score changes were observed in the two groups of patients with intra-tracheal intubation 1,3 and 7 days after VAP onset.Comparisons of CPIS and PCT scores were carried out between two groups at different intervals after VAP onset.Data were statistically processed by SPSS 19.0.Count data were tested by x2,and measurement data were expressed by mean ± standard deviation (x-±s).The comparison within the groups was made by t test,whereas the comparison between the groups was by means of repeated measure analysis of variance and Bonferroni test;Pearson linear correlation analysis was used,and P ＜ 0.05 was considered statistically significant.Results At the same observation interval in two groups of patients with VAP,there was no significant difference in the serum levels of PCT and CPIS score (P ＞ 0.05);there were significant differences in PCT and CPIS scores between VAP occurred at the first day,the third day and at time of intra-tracheal intubation in two groups (P ＜ 0.05).However,comparison of PCT and CPIS scores at the seventh day,the differences were not statistically significant (P ＞ 0.05).In addition,at the 7 days the correlation coefficient between PCT and survival patients' mechanical ventilation,days in ICU and total length of hospital stay were 0.92,0.83,and 0.71,respectively,yet the 7-day CPIS score correlation coefficients were 0.83,0.74 and 0.70,(both P ＜ 0.05).Conclusions Early monitoring of serum PCT and CIPS score of pulmonary tuberculosis patients can judge the incidence of VAP,and the variations of PCT and CIPS score can predict the severity and prognosis of the disease as well.

OBJECTIVE: To discuss the clinical features, diagnosis and endoscopic surgical intervention for small steoma of nasal sinuses causing nasal and facial pain. METHOD: A retrospective review was performed on 21 patients with nasal and facial pain caused by small osteoma of nasal sinuses, and nasal endoscopic surgery was included in the treatment of all cases. RESULT: The nasal and facial pain of all the patients was relieved. Except for one ase exhibiting periorbital bruise after operation, the other patients showed no postoperative complications. CONCLUSION Nasal and facial pain caused by small osteoma of nasal sinuses was clinically rare, mostly due to the neuropathic pain of nose and face caused by local compression resulting from the expansion of osteoma. Early diagnosis and operative treatment can significantly relieve nasal and facial pain.
Endoscopy ; Face ; Facial Pain ; Humans ; Nasal Surgical Procedures ; Osteoma ; surgery ; Paranasal Sinuses ; pathology ; Postoperative Complications ; Retrospective Studies

OBJECTIVETo investigate the distribution and drug-resistance of bacteria in the lower respiratory tract in patients with tuberculosis and severe pneumonia receiving invasive mechanical ventilation.

METHODSThe clinical data, lower respiratory tract infection pathogens and bacterial drug sensitivity were analyzed in 208 patients receiving invasive mechanical ventilation for tuberculosis and severe pneumonia.

RESULTSA total of 355 pathogenic microbial strains were obtained from the patients, among which 281 (79.2%) strains were Gram-negative bacteria, 62 (17.5%) were fungi, and 12 (3.4%) were Gram-positive bacteria. Mixed infections were found in 68 cases (19.2%). The sensitivity rates of meropenem, imipenem and amikacin were over 60% for Gram-negative bacteria, and those of teicoplanin, vancomycin, and fusidic acid were 100% for Gram-positive bacteria.

CONCLUSIONThe main pathogenic bacteria are Gram-negative bacteria, fungi and Gram-positive bacteria in the lower respiratory tract of patients with tuberculosis and severe pneumonia receiving mechanical ventilation. Meropenem, imipenem and amikacin are effective antibiotics for lower respiratory tract infections, and multi-drug resistance is frequent in these patients, which urges appropriate use of the antibiotics.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Gram-Negative Bacteria ; isolation & purification ; Gram-Positive Bacteria ; isolation & purification ; Humans ; Microbial Sensitivity Tests ; Pneumonia, Bacterial ; microbiology ; Respiration, Artificial ; Respiratory Tract Infections ; microbiology ; Tuberculosis ; microbiology