Objective To investigate the expression and purification I278T-mutant human cystathionineβsynthase(CBS) in E . coli .Methods Site-directed mutagenesis by overlap extension using the polymerase chain reaction (PCR) was employed to construct mutant plasmids pGEX4T-1-CBS(I278T) ,which was induced and expressed in a medium containing 3% ethanol ,purified by affinity chromatography to obtain mutated CBS (I278T) protein .The activity ,UV-visible absorption spectroscopy ,protein particle size and Zeta potential of the purified protein were measured .Results Plasmid pGEX4T-1-CBS(I278T) was successfully constructed .The yield ,the specific activity and activity recovery of purified mutant CBS (I278T ) protein were 2 .3 mg/L ,21 .4 U/mg and 22 .6% .S-adenosylmethionine(AdoMet) with final concentration of 1 mmol/L showed no activation toward mutant CBS (I278T) protein .Ac-cording to UV-visible absorption spectroscopy analysis ,purified mutant CBS(I278T) had characteristic absorption peaks at 429 nm and 550 nm for heme-binding proteins .Protein average particle size was 7 .5 -10 .1 nm ,mainly in the form of tetramers ,and Zeta potential was - 16 .3 mV .Conclusion The methods of expression ,purification and identification of I278T-mutant human cystathionineβsynthase in E .coli were successfully established .

Objective To evaluate interleukin-27 ( IL-27 ) as a sepsis diagnostic biomarker in critically ill adults with sepsis. Methods A retrospetive study was conducted. A total of 176 systemic inflammatory response syndrome ( SIRS ) patients in Department of Critical Care Medicine of Xinxiang Medical College First Affiliated Hospital from March to November in 2014 were enrolled. The patients were divided into no sepsis group ( n=66 ), pulmonary originated sepsis group ( n=65 ), and non-pulmonary originated sepsis group ( n=45 ). Plasma IL-27 and procalcitonin ( PCT ) were determined with enzyme linked immunosorbent assay ( ELISA ). Receiver operating characteristic curve ( ROC ) and classification and regression tree methodology was used to evaluate diagnostic biomarker performance. Results The proportion of patients in pulmonary original sepsis group whose body temperature in line with SIRS criteria was significantly higher than no sepsis group ( 66.2%vs. 44.5%, P<0.05 ), and they were easy to suffer from tumor ( 44.6%vs. 22.7%, P<0.05 ). The proportion of patients in non-pulmonary originated sepsis group whose white blood cell count in line with SIRS criteria was significantly higher than no sepsis group ( 68.9%vs. 42.7%, P<0.05 ). It indicated that patients in pulmonary originated sepsis group and non-pulmonary originated sepsis group were more in line with SIRS criteria compared with no sepsis group. It was shown by ROC curve that IL-27 and PCT was not effective in discriminating sepsis among unselected patients showing symptoms and signs of SIRS. The area under the curve ( AUC ) was 0.59 [ 95%confidence interval ( 95%CI )=0.49-0.65 ] and 0.61 ( 95%CI=0.55-0.71 ). According to the further analysis from different infection sources, the highest AUC was 0.71 ( 95%CI=0.59-0.79 ) for IL-27 in patients with a non-pulmonary originated sepsis. A decision tree incorporating IL-27, PCT, and age had an AUC of 0.78 ( 95%CI = 0.71-0.87 ) in patients with a non-pulmonary originated sepsis, which was higher than IL-27 [ 0.71 ( 95%CI = 0.59-0.79 ) ] or PCT [ 0.65 ( 95%CI = 0.57-0.78 ) ]. Compared to that of pediatric cohort with sepsis, lower expression of IL-27 was found in adult patients. Conclusions IL-27 performed overall poorly as a sepsis diagnostic biomarker in adults. IL-27 may be a more reliable diagnostic biomarker for sepsis in children than in adults. The combination of IL-27 and PCT can reasonably estimate the risk of sepsis in subjects with a non-pulmonary originated sepsis.

Objective To analyze the recurrence rate of intubation and increase of ventilator support rate within 24 hours after using fiberoptic bronchoscopy (FOB) in critically ill patients with hypoxemia complicated with respiratory failure, and to approach the feasibility of FOB in such patients.Methods A prospective study was conducted, including 200 critically ill patients with acute respiratory failure using FOB [oxygenation index (PaO2/FiO2) ≤ 300 mmHg (1 mmHg = 0.133 kPa)] admitted to the intensive care unit (ICU) of the First Affiliated Hospital of Xinxiang Medical College. The rates of intubation and increased ventilatory support and the reasons for bronchoscopy related complications after using FOB 24 hours were recorded, the main risk factors leading to these changes and complications were analyzed and screened by logistic regression analytic method.Results Within 24 hours after using FOB for 200 patients with respiratory failure, an increase in mechanical ventilatory support was required in 68 patients (34%) of that 28 (14%) led to endotracheal intubation. With the extension of time, the rates of intubation and ventilatory support showed a tendency of elevation, the rise in ventilatory support rate being faster. The reasons for bronchoscopy related complications after FOB consisted of cardiovascular disease (41%), coronary artery disease (17%), chronic obstructive pulmonary disease (COPD, 17%), chronic restrictive pulmonary disease (10%), immunity suppression (54%), malignant neoplastic hematologic disorder (20%), acquired immune deficiency syndrome (AIDS, 12%), solid organ transplantation (3%), solid tumor (10%), corticosteroid therapy (25%), immunosuppressive drug (16%), diabetes (15%), chronic renal failure (14%), swallowing nerve injury (37%), anticoagulant therapy (19%), antiplatelet therapy (13%). In the patients with occurrence of COPD or immunosuppression, the rate of invasive ventilation used was significantly higher than that without using invasive ventilation [COPD: 35% (10/28) vs. 14% (24/172),χ2 = 8.081,P = 0.004; immunosuppression: 75% (21/28) vs. 50% (86/172),χ2 = 6.051,P = 0.014]. The logistic regression analysis showed that the occurrence of COPD or immunosuppression was obviously related to whether the intubation being necessary or not [COPD: odds ratio (OR) = 5.200, 95% confidence interval (95%CI) = 1.500 - 17.700,P = 0.006; immunosuppression:OR = 5.300, 95%CI =1.600 - 17.100,P = 0.004].Conclusions In patients with hypoxemia using FOB, they often require addition of mechanical ventilatory support, but the intubation rate is not high. Under the ventilatory support, FOB has certain feasibility for treatment of critically ill patients with hypoxemia and acute respiratory failure.

OBJECTIVE:To construct the structure equation model for the burden of chronic disease inpatients in primary hos-pital before and after the implementation of zero price policy,and analyze the changes of burden factor loading. METHODS:6 pri-mary hospitals were randomly selected in Danyang,Jiangsu province. The data of hospitalization expenses for chronic diseases(hy-pertension,diabetes,bronchitis)were collected before and after the implementation of zero price policy. Using drug cost,nursing fees,inspection fees and treatment fees as independent variables,hospitalization burden as latent variable,SPSS and AMOS 24.0 software were adopted to establish the model. RESULTS:The burden factor loading of inpatients changed greatly before and after the implementation of zero price policy. χ2=24.586,χ2/df=1.446, RMSEA=0.019,GFI=0.995,AGFI=0.989,CFI=0.988,NFI=0.963 manifested good model fitting. Factor loading of drug cost increased greatly,indicating the burden of inpatients was reduced after the implementation of zero price policy. Factor loading of inspection fees and treatment fees increased significantly,the charac-terization effects of them to the burden of inpatients were enhanced. Factor loading of nursing fees was the lowest,and it had the weakest effects on the burden of inpatients,of which the government should enhance the regulation. CONCLUSIONS:Established model has good reliability and validity. It can reflect the change of burden factor loading of inpatients before and after the implemen-tation of zero price policy.

Objective To explore the relation of stroke and metabolic syndrome (MS), and provide theoretical evidence for prevention of stroke and MS. Methods Five hundred stroke patients accompanied with MS were enrolled, and divided into different types according to the distribution of the components of metabolic syndrome and abnormal metabolic condition. The systolic blood pressure(SBP) ,diastolic blood pressure( DBP),fasting glycemia ( FPG ), triglycerides ( TG ), total cholesterol ( TC ), low density density lipoprotern-cholesterol (LDL-C), high density density lipoprotern-cholesterol (HDL-C)of each patient were detected and recorded. Results Among these stroke patients with MS, most patients were 60 - 69 years old ( 39.60% ), and there was no significant difference between two gender( P = 0. 545 ). Among each component of MS, hypertension was most related to MS;patients complicated with hypertension + hyperlipidemia + hyperglycemia were the most major proportion of stroke patients with MS(48. 00% ). Conclusion MS and its components are all risk factors for stroke,and the more complication the patients had,the more patients suffered from stroke with MS. Targeted interventions of different type of abnormal condition has important prevention effect on MS and stroke.

To prove the influence of protein isoaspartyl-methyltransferase ( PIMT ) on the cell apoptosis of fibroblast-like synoviocytes of RA.Methods: The expression vector of PIMT was constructed and transfected in to the RA-FLS, the impact of PIMT on the cell apoptosis of RA-FLS was observed by overexpressing the vector of PIMT.Results:The mRNA and protein level of PIMT in RA-FLS was increased after transfected the vector of PIMT into RA-FLS;compared with the normal cultured RA-FLS and the RA-FLS transfected with the empty vectors ,the cell apoptosis level was also increased.Conclusion:The decreased expression level of PIMT in RA-FLS is an important reason for reduce apoptosis of RA-FLS,and PIMT can affect the imbalance of proliferation/ap-optosis in the RA-FLS.

Objective To assess the value of color Doppler ultrasonogarphy in the diagnosis of subacute thyroiditis (SAT). Methods Sixty-eight patients of SAT and 30 patients with thyroid papillary carcinoma were included, and their imaging features were retrospectively reviewed. Results All the patients with SAT had either focal or diffuse hypoechogenicity of the thyroid lobes with rich blood supplying in the diseased region, among them 52 patients (76.47%, 52/68) had tenderness on palpation. Diffuse and focal lesions were found in 40 (58.82%, 40/68) and 28 patients (41.18%, 28/68). The boundary and shape of focal lesions had no diagnostic characteristics compared with those of thyroid papillary carcinoma. Punctate calcification and resistance index (RI) in the nodule were significant different between the SAT and thyroid papillary carcinoma (P＜0.05). Conclusion Color Doppler ultrasonography can display the features of SAT and is an effective method for the diagnosis of SAT.

Objective To investigate the diagnostic value of bronchoscopy in smear negative pulmonary tuberculosis.Methods 65 cases with cavernous and 35 non -cavernous negative smear pulmonary tuberculosis were examined by bronchoscopy.Results The positive rates of sputum examination in the empty group,the non empty group and the total sputum examination were 49.2%,17.1% and 38%,respectively.There was statistically signifi-cant difference in sputum positive rate between empty group and non empty group (χ2 =9.942,P =0.002).The posi-tive rate of sputum was 28%.The positive rate of sputum negative and sputum positive rate was 10% after operation. Microscopically visible tracheal and bronchial abnormalities,tracheal inflammatory change rates of empty group and non empty group were 52.3%,40.0%,the difference between the two groups had no statistically significant difference (χ2 =1.381,P =0.240).The tracheal stenosis rates of hole group and non empty group were 9.0% and 11.4%,the difference between the two groups had no statistically significant difference (χ2 =0.122,P =0.727).The overall trachea abnormal changes rate was 58.0%,including inflammatory change rate 48.0% and 10.0% stenosis rate. Conclusion Bronchoscopy has significant clinical value in smear negative pulmonary tuberculosis,especially with cavernous.The positive rate will be promoted after bronchoscopy,and can find abnormal changes of tube.

Objective: To analyze the relationship between cognitive function and tumor necrosis factor receptor 1 (TNFR1) expression of occupational exposed workers to aluminum and provide the basis for the diagnosis of cognitive impairment. Methods: 140 cases Shanxi aluminum plant workers were collected in 2016 as the research object, including 70 potroom workers for exposure group, 70 non-electrolytic aluminum plant workers in the control group, respectively. Using mini mental status examination (MMSE), digit span test (DST), fuld object memory examination(FOME) and simple reaction time test(SRTT) evaluate the cognitive function of objects. Using graphite furnace atomic absorption method for the determination of plasma aluminum levels as an indicator of aluminum contact exposure of workers. Using RT-PCR method for detection of tumor necrosis factor receptor 1 (TNFR1) mRNA expression levels. And comparison group differences in cognitive and TNFR1 mRNA expression levels. Results: The plasma aluminum content of exposed group (77.12±27.18) μg/L higher than the control group (55.6±28.69)μg/L (P=0.000); Compared to control group, FOME and MMSE score was significantly increased in the exposed group (P=0.000, P=0.000), SRTT scores significantly higher in the exposed group (P=0.001), DST no significant difference in the control group (P=0.893). Compared to control group, The mRNA expression of TNFR1 was significantly higher in the exposed group(P=0.002); Compared to control group, The protein expression of TNFR1 was significantly higher in the exposed group (P=0.002). By correlation analysis in exposure group, plasma aluminum content was negatively correlated with MMSE and the DST (r=-0.284, r=-0.331, P<0.05) and positively correlated with the SRTT, TNFR1 (mRNA) and TNFR1(protein)(r=0.255, r=0.333, r=0.987, P<0.01), MMSE was negatively related to TNFR1 (mRNA) and TNFR1 (protein) (r=-0.268, r=-0.255, P<0.05); DST was negatively correlated with the SRTT and TNFR1 (protein)(r=-0.267, r=-0.330, P<0.05); SRTT was positively correlated with TNFR1 (protein)(r=0.243, P<0.05); TNFR1 (mRNA) was positively correlated with TNFR1 (protein)(r= 0.340, P<0.01). Conclusion Cognitive function change of occupational exposed workers to aluminum was related to the increase of TNFR1 expression.

OBJECTIVE: To study the genotype and phenotype of fetuses with 22q11.2 microdeletion and other abnormalities such as cardiac malformation and cleft palate. METHODS: Fetal ultrasound was carried out at 12 weeks to 20 to 24 weeks of gestation. After excluding the chromosomal karyotype abnormality, single nucleotide polymorphism (SNP) array was used to detect copy number variations of 5 fetuses with heart development abnormality or other structural abnormalities. The fetus with 22q11.2 microdeletion and its parents were also subjected to multiplex ligation-dependent probe amplification (MLPA) assay. RESULTS: SNP array analysis showed that the 5 fetuses had all carried a 2.27-3.02 Mb deletion of the 22q11.2 region. MLPA assay confirmed that LCR22-A-B was involved in all cases, and that all deletions were de novo in origin. CONCLUSION It is of great significance to exclude 22q11.2 microdeletions in fetuses with cardiac malformations. The deletion regions of these fetuses are similar but different, and the phenotypic difference is related to their genotypes.