The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

The effect of magnetic Fe3O4 particles on cellulase in the enzymatic hydrolysis of sunflower seed hull was studied in different adding ways and additive amount. In the process of enzymatic hydrolysis of sunflower seed hull, the variations of cellulase activity, reducing sugar concentration and cellulose conversion were evaluated. After the reaction, the analysis of pH and surface tension of hydrolysate were also used to determine the mechanisms of cellulase by the magnetic effect. The results indicated that after adding magnetic Fe3O4, the cellulase activity, reducing sugar concentration and conversion of cellulose had an increased between the 0.5 g/L and 2.0 g/L cases after 48 h. When the additive amount of magnetic Fe3O4 was 2 g/L, the cellulase activity at 60 h was improved significantly by 25.9%. It was found that the concentration of reducing sugar was increased from 6.950 mg/mL to 8.775 mg/mL with magnetic Fe3O4 1.5 g/L. Simultaneously, compared with the blank, which the conversion of cellulose was 47.932%, the maximum celluloseconversion of samples with adding magnetic Fe3O4 was 60.531%. Besides, the stability of cellulase activity adding in times was better than in one time. After the reaction, the final surface tension of hydrolysate with 1.5 g/L magnetic Fe3O4 was the lowest in comparison with the blank. However, no significant differences were observed in the final pH of the hydrolysate.

By using whole blood selenium, 24 hr urinary selenium and hair selenium contents as the indices of assessing human selenium status, it was found that the populations in the endemic areas of Keshan disease were practically in a selenium poor status. The selenium contents in locally grown staple grains and daily diets in the endemic areas were also lower than those in the non-endemic areas. In an area covering a cross section of Keshan disease geographic belt in our country, the hair selenium contents of agricultural populations were measured. The results indicated that all the hair selenium contents in the endemic sites were always at a lower level, whereas those in the non-endemic sites distant from the endemic areas were generally at a higher level; they decreased gradually until the endemic areas were reached; and finally, along the contiguous region of the endemic and non-endemic areas they were insignificantly different.The hair selenium contents among the agricultural populations were significantly lower than those among the non-agricultural ones in the same endemic areas. However, no regular correlation had been observed between the seasonal prevalence of Keshan disease and the variation of hair selenium contents in the same populations living in the same endemic sites.It is considered that the endemic areas of the disease seem to be a Se-deficiency belt, and Se-deficiency probably might be a pathogenic geo-gen in the prevalence of Keshan disease.

Objective:To explore the effects of fluorescence method and improved expansion and collapse method on cardiopulmonary function, immune function and safety during segmental pneumonectomy.Methods:One hundred and fifty-six patients with lung segmental resection operation in Yucheng People′s Hospital from March 2018 to August 2020 were enrolled. They were divided into two groups according to the random number table method, each group with 78 patients. The patients in the observation group were treated with fluorescence method and the patients in the control group were treated with modified expansion collapse method. The operation indexes, immune function and cardiopulmonary function of the two groups were compared, and the incidence of postoperative complications were counted.Results:The intraoperative time and the appearance time of intersegmental plane in the observation group were shorter than those in the control group: (105.33 ± 10.62) min vs. (139.46 ± 12.58) min, (15.46 ± 1.13) s vs. (724.56 ± 65.65) s, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of 6 min walking distance, peak expiratory flow rate, percentage of forced vital capacity and percentage forced expiratory volume in 1 second in the observation group were higher than those in the control group: after operative for 1 d: (475.36 ± 10.29) m vs. (412.11 ± 13.33) m, (284.52 ± 10.33) L/min vs. (251.13 ± 12.89) L/min, (85.65 ± 3.21)% vs. (81.13 ± 1.43)%, (83.25 ± 2.55)% vs. (74.49 ± 2.53)%; after operative for 7 d: (510.23 ± 16.66) m vs. (488.33 ± 15.42) m, (302.13 ± 15.58) L/min vs. (285.12 ± 10.22) L/min, (93.46 ± 5.79)% vs. (88.44 ± 5.44)%, (92.25 ± 2.32)% vs. (85.54 ± 2.13)%, there were statistical differences ( P<0.05). After operative for 1, 7 d, the levels of CD 4+/CD 8+, CD 8+, CD 4+ in the observation group were higher than those in the control group: after operative for 1 d: 0.85 ± 0.10 vs. 0.52 ± 0.04, 0.305 ± 0.025 vs. 0.285 ± 0.012, 0.325 ± 0.021 vs. 0.304 ± 0.025; after operative for 7 d: 1.13 ± 0.10 vs. 1.02 ± 0.07, 0.324 ± 0.029 vs. 0.306 ± 0.023, 0.359 ± 0.024 vs. 0.332 ± 0.025, there were statistical differences ( P<0.05). The rate of postoperative complications in the observation group was lower than that in the control group: 2.56%(2/78) vs. 11.54%(9/78), there was statistical difference ( χ2 = 4.79; P = 0.029). Conclusions:Both the fluorescence method and the modified inflation-collapse method have certain therapeutic effects, but the combination of fluorescence method in segmentectomy can provide reliable technical support and reduce the impact on cardiopulmonary function.

Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.
Cell Proliferation ; Genes, Bacterial ; physiology ; Lipopolysaccharides ; biosynthesis ; Mutation ; Polysaccharides, Bacterial ; biosynthesis ; Xanthomonas campestris ; genetics

OBJECTIVETo discuss the causes of unsatisfied cervical posterior decompression surgery and describe the overhauling strategies and precaution.

METHODSThe clinical data of 14 patients required revision surgery were retrospectively analyzed, and these patients with unsatisfied effects were due to cervical posterior decompression surgery from January 2012 to December 2014. Overhauling reasons were analyzed and then different revision procedures were performed. The functions of cervical cord and ambulation were evaluated respectively by modified Japanese Orthopedic Association(mJOA) score and Nurick grade according to the course order:preoperative for the first time, pre-revision and at final follow-up. Improvement rate of nerves function were calculated before and after operation for the first time, before and after revision. Above data were statistically analyzed by SPSS16.0 software.

RESULTSReoperation reasons including 2 patients with the insufficiency width of laminectomy, 2 patients with the inadequate length of decompression, 2 patients with nerve root and spinal cord compression caused by fractured collapse, 4 patients with closed the door of vertebral lamina, 1 patient with less open-door angle, 2 patiens with ossification of posterior longitudinal ligament (1 case complicated with close the door), 2 patients with cervical spine kyphotic deformity aggravating (1 case complicated with close the door), 1 patient with nerve root canal stenosis caused by uncovertebral joint hyperplasia. Preoperative for the first time, pre-revision and at final follow-up, mJOA scores were 11.89±1.67, 13.11±1.09, 15.61±0.59, and Nurick grades were 4.21±0.58, 3.57±0.51, 1.71±0.47, respectively. There was significant difference between final follow-up and preoperative for the first time, pre-revision(<0.05). Improvement rate of nerve function was (22.33±9.49)% with bad before and after operation for the first time, and (64.60±9.88)% with good before and after revision, with statistical significance(<0.05).

CONCLUSIONSIndividualized revision surgery based on different causes for unsatisfied cervical posterior decompression can improve the function of spinal cord. Preoperative carefully analyzing the etiological factors, thoroughly decompression can reduce the revision rate.

Xanthomonas campestris pv. campestris ( Xcc), causative agent of the black rot disease of cruciferous crops worldwide, produces large amount of extracellular polysaccharide( EPS), which has found wide applications in industry. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5gus A5, and a number of EPS-defective mutants were isolated. The Tn5gusA5 insertion sites in the mutants were analyzed by using thermal asymmetric interlaced PCR(TAIL-PCR), and the corresponding genes were identified by homology blast to the completely sequenced genome of Xcc 8004 strain. A novel gene, waxE, identified from the EPS-defective mutant 151D09, was found to be disrupted by the insertion of Tn5gusA5 in the open reading frame(ORF) with genome coordinates 4478998bp to 4479819bp.This gene showed 52% similarity to the kdtX gene of Serratia marcescens and 50% to the waaE of Klebsiella pneumoniae at amino acid level, with characteristics of glycostransferase 2 family domain. In order to identify the function of waxE gene, waxE gene deletion mutant of Xcc 8004 was constructed by gene replacement strategy in which waxE gene of genome was replaced by kanamycin resistant gene kan. The waxE gene deletion mutant strain, named Xcc 8570, was confirmed by both PCR and southern analysis. The growth rate of the deletion mutant 8570 in rich medium was not affected, but the EPS yield reduced by 35% as compared with the wildtype strain 8004. The deletion mutant could be completmented in trans with plasmid pLATC8976 harboring an intact waxE gene, and the EPS yield of the mutant was restored. The combined data showed that waxE gene involved in EPS biosynthesis in Xcc.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; genetics ; metabolism ; Blotting, Southern ; Cloning, Molecular ; DNA Transposable Elements ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polysaccharides, Bacterial ; genetics ; metabolism ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Xanthomonas campestris ; genetics ; metabolism

BACKGROUNDThe prevalence of HIV/AIDS in Chinese ethnic minorities is an important component of China's AIDS issues. In this study, we launched an intervention programme in Yunnan Province of China, where the Dai people live, to carry out the community-based HIV/AIDS health education and behavioral interventions on ordinary Dai farmers. The Dai people believe in Theravada Buddhism.

METHODSFour rural communities were randomly divided into two groups. In one group (Buddhist group), HIV/AIDS health education and behavioral intervention were carried out by monks. The other group (women group) was instructed by women volunteers. The intervention continued for one year and the data were collected before and after the intervention project.

RESULTSIn the Buddhist group, the villagers' AIDS related knowledge score was boosted from 3.11 to 3.65 (P < 0.001), and some indices of the villagers' behavior using condoms improved after the intervention. But this improvement was poorer than that in the women group. In the Buddhist group, the villager's attitude score towards the people living with HIV and AIDS (PLWHA) also increased significantly from 1.51 to 2.16 (P < 0.001).

CONCLUSIONThe results suggested that the Buddhist organization has limited success in promoting the use of condoms, but plays an important role in eliminating HIV/AIDS related discrimination.

Acquired Immunodeficiency Syndrome ; prevention & control ; Adolescent ; Adult ; Buddhism ; China ; Condoms ; Female ; HIV Infections ; prevention & control ; Health Education ; methods ; Humans ; Male ; Middle Aged ; Young Adult

BACKGROUND: This study aimed to determine whether modified shock index (MSI) is associated with mortality that is superior to heart rate, blood pressure, or the shock index (SI) in emergency patients.METHODS: A retrospective database review was performed on 22161 patients who presented to Peking Union Medical College Hospital Emergency Department and received intravenous fluids from January 1 to December 31, 2009. We gathered data of the patients on age, gender, vital signs, levels of consciousness, presenting complaints, and SI and MSI were calculated for all patients.RESULTS: Multivariate regression analysis was performed to determine the correlation between risk factors and outcome. There is a significant correlation between emergency patient mortality rate and patient's vital signs obtained at the triage desk (HR>120 beats/min, systolic BP<90 mmHg, diastolic BP<60 mmHg). MSI is a stronger predictor of emergency patient mortality compared to heart rate and blood pressure alone, whereas SI does not have a significant correlation with emergency patient mortality rate.CONCLUSION: MSI is a clinically significant predictor of mortality in emergency patients. It may be better than using heart rate and blood pressure alone. SI is not significantly correlated with the mortality rate of the emergency patient.

Objective Toexploretheprognosticvalueofpreoperativeneutrophil-to-lymphocyteratioandderived neutrophil-to-lymphocyteratioincolorectalcancer(CRC)individuals.Methods Theclinicalpathologicaldataand preoperative blood routine test results were collected from medical records,and 5 year follow up was performed in a total of 555 surgically resected CRC cases.Receiver operative curve (ROC)was used to calculate NLR and d -NLR cut-off value,and Kaplan-Meier curve and multiple COX regression were selected to evaluate the influence of preoperative NLR and d -NLR on clinical outcome of CRC cases and prognostic predictive nomogram was established to evaluate the predictivevalueofNLRandd-NLR.Results Usingoverallsurvival(OS)asanendpoint,theoptimalcut-offvaluesof NLR and d-NLR were 3.21 (Sensitivity=0.752,specificity=0.753,AUC=0.762)and 2.12 (sensitivity=0.721, specificity=0.683,AUC=0.720),respectively.Preoperative NLR and d-NLR were significantly associated with free-recurrent survival (RFS)and OS(P<0.01).NLR and d-NLR were independent factors for prediction of RFS (HRNLR=2.53,HRd-NLR=1.60)and OS (HRNLR=2.75,HRd-NLR=2.11)in II-III stage preoperative CRC patients.The C-indexes of RFS and OS predictive nomograms including NLR and d-NLR were 0.851 and 0.836,and C-indexes without NLRandd-NLRwere0.801and0.793,respectively.Conclusion ThisresultsindicatedthatRFSandOSofthe patients with preoperative high NLR and d-NLR were significantly shorter than those with relative low NLR and d-NLR, and they were independent prognostic predictive factors for RFS and OS,and nomograms including NLR and d-NLR could significantly improve the prognostic predictive efficacy in postoperative CRC individuals.