Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

Nasal polyp (NP) is a common chronic inflammatory disease of the nasal cavity and sinuses.Although some authors have suggested that NP is related to inflammatory factors such as interleukin (IL)-1β,IL-5,IL-8,granulocyte-macrophage colony-stimulating factor (GM-CSF),tumor necrosis factor (TNF)-α,and IL-17,the mechanisms underlying the pathogenesis and progression of NP remain obscure.This study investigated the expression and distribution of IL-17 and syndecan-1 in NP,and explored the roles of these two molecules in the pathogenesis of eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) and non-Eos CRSwNP.Real-time PCR and immunohistochemistry were used to detect the expression of IL-17 and syndecan-1 in samples [NP,unciform process (UP) from patients with CRS,and middle turbinate (MT) from healthy controls undergoing pituitary tumor surgery].The results showed that the expression levels of IL-17 and syndecan-1 were upregulated in both NP and UP tissues,but both factors were higher in NP tissues than in UP tissues.There was no significant difference in IL-17 levels between the Eos CRSwNP and non-Eos CRSwNP samples,and syndecan-1 levels were increased in the non-Eos CRSwNP tissues as compared with those in Eos CRSwNP tissues.In all of the groups,there was a close correlation between the expression of IL-17 and syndecan-1 in nasal mucosa epithelial cells,glandular epithelial cells,and inflammatory cells,suggesting that IL-17 and syndecan-1 may play a role,and interact with each other,in the pathogenesis ofnon-Eos CRSwNP.

Objective To investigate iodine deficiency disorders(IDD) in Chongqing and Linzhi, and to provide scientific basis for IDD control and prevention. Methods According to the national program developed in 2007, investigation was conducted in Chengkou and Wuxi county in Chongqing municipality, and Linzhi, Bomi,Milin and Langxian county in Linzhi prefecture. Five towns were sampled in Linzhi county, and 3 in other counties.In each town, one township primary school and two village primary schools were selected to inspect thyroid by B ultrasound and palpation, and urinary iodine of children aged 8 to 10 years was tested in these schools. Meanwhile,2 villages were selected in each town for test of salt iodine level and urinary iodine of childbearing age women and search cretin cases. Results Three hundred and forty families in Chongqing and 915 families in Linzhi were investigated. The coverage of iodized salt in Chongqing was 98.82%(336/340), which was significantly higher than that in Linzhi[66.34%(607/905), x2 = 139.56, P ＜ 0.01]. Goiter rate of children in Chongqing was 9.27%(89/960) by palpation and 8.34% (61/731) by B ultrasound, while goiter rate of children in Linzhi was 7.80%(102/1308) by palpation and 5.53% (69/1248) by B ultrasound. The difference of goiter rate by palpation between Chongqing and Linzhi was not statistically significant (x2 = 1.37, P ＞ 0.05 ). But goiter rate of children by B ultrasound in Chongqing was higher than that in Linzhi (x2= 5.51, P ＜ 0.05). In Chongqing, the median urinary iodine was 319.15 μg/L, and 345.75 μg/L in Chengkou county and 281.39 μg/L in Wuxi county. In Linzhi prefecture, the median urinary iodine was 189.81 μg/L, and 207.81 μg/L in Linzhi county, 161.12 μg/L in Bomi county, 131.83 μg/L in Milin county and 334.60 μg/L in Langxian county. The median urinary iodine in childbearing women were 248.42 μg/L in Chongqing and 121.25 μg/L in Linzhi. The median urinary iodine in Chongqing both in children and women were higher than those in Linzhi. No new cretin case was found in these two areas. Conclusions Goiter rate in high risk areas of IDD in Chongqing and Linzhi has decreased to less than 10%.No new cretin case is found in these areas. It can be concluded that the work of control and prevention is effective.There is excess iodine in Chongqing. In Linzhi county and Langxian county, iodine is excess in children and deficient in women. Further investigation should be conducted to find out the reason. Population iodine is excess in Bomi and Milin counties. The concentration of salt iodine should be decreased in Chongqing. In Linzhi prefecture,adding iodine measures should be adjusted based on further investigation.

OBJECTIVE: To observe the changes of cardiac functional indicators in spontaneously hypertension rats (SHR) suffered by struck injuries of soft tissues, form which to discuss the significance in relationship between injury and disease. METHODS: SHRs were divided into four groups according to the degree, and whose thigh was struck with the homemade hit instrument. Then the blood pressure, heart rate as well as serum biochemical indicators (CK, CK-Mb, LDH, Cr, BUN, HBDH and AST) were examined. In addition, the morphological changes of heart, brain and kidney were also observed. RESULTS: The observe values of heart rate and blood pressure of SHRs seemed have relation to the degree of damage, but there were no statistically significance between different hit degree groups. In terms of serum biochemical indicators, all seven indicators of the hit group had significant difference with that of the control group, and the AST CK, CK-MB levels were difference between the group of different strike degree. CONCLUSIONS Struck injuries of soft tissues can result in forepart damage of cardiac muscle. The biochemical indicators reflect the degree of injury, and they could be used as a type of objective enzymology indicators for analysis of the relationship between injury and disease in forensic practice.
Animals ; Aspartate Aminotransferases/blood* ; Biomarkers/blood* ; Blood Pressure ; Creatine Kinase/blood* ; Female ; Heart/physiopathology* ; Heart Rate ; Hindlimb/injuries* ; Hypertension/physiopathology* ; Kidney/physiopathology* ; Male ; Myocardium/pathology* ; Random Allocation ; Rats

Coronary Vessel Anomalies ; diagnosis ; Electrocardiography ; Humans ; Infant ; Pulmonary Artery ; abnormalities

OBJECTIVETo evaluate the biological characteristics of Campylobacter jejuni (CJ) cultured on different culture media and their expression abundance of outer membrane proteins (OMPs).

METHODSCJ was cultured on the improved Bull's medium yolk agar, improved Bull's blood agar or improved Bull's agar for 48 h. The biological characteristics of the bacteria, including the colony feature, morphology, motility, biochemistry, and results of indirect fluorescence test were observed and compared. OMP of the cultured CJ was extracted using 0.2 mol/L and glycine-hydrochloride buffered solution (pH 2.2) and identified by SDS-PAGE to compare the expression abundance of the OMPs with molecular weight of 28-31 kD.

RESULTSCJ exhibited typical biological characteristics with larger cell body and more rapid growth on improved Bull's medium yolk agar than those on improved Bull's blood agar and improved Bull's agar. The bacteria grown on improved Bull's medium yolk agar showed also greater expression abundance of the OMPs with molecule mass between 28 kD and 31 kD.

CONCLUSIONImproved Bull's medium yolk agar allows rapid growth of CJ with typical biological characteristics and enhanced expression of the OMPs with molecular weight of 28 -31 kD, and can be widely used in CJ subunit vaccine development, CJ epidemiological survey, CJ food safety examination, and CJ quarantine.

Bacterial Outer Membrane Proteins ; analysis ; metabolism ; Campylobacter jejuni ; growth & development ; metabolism ; Culture Media

OBJECTIVETo compare primary anastomosis after intraoperative colonic defecation and Hartmann procedure for obstructive left colon cancer.

METHODSClinical data of 68 patients who underwent emergent laparotomy for left colon cancer with acute bowel obstruction between January 2000 and January 2008 were analyzed retrospectively.

RESULTSPrimary resection and anastomosis with intraoperative defecation was performed in 43 patients and Hartmann's procedure in 25 cases. Patients in both groups were comparable in terms of age, gender, nutritional status, underlying diseases, tumor location and stage, etc. The morbidity and mortality in the two groups were 25.6% vs 28.0% (P=0.761) and 2.3% vs 4.0% (P=0.369), respectively, and the differences were not statistically significant. The length of hospital stay (including first resection operation and second admission for colostomy closure) was (16.6+/-7.8) d in the primary anastomosis group and (24.6+/-9.4) d in the Hartmann procedure group, and the difference was statistically significant (P=0.002). The costs of hospitalization in the two groups were CNY 50,192.8+/-39,727.4 and CNY 58,382.1+/-30,304.9 (P=0.020).

CONCLUSIONPrimary resection with intraoperative colonic defecation is safe and effective, and should be considered as an alternative to Hartmann procedure for obstructive left colon cancer in selected patients.

Aged ; Anastomosis, Surgical ; methods ; Colectomy ; methods ; Colon ; surgery ; Colonic Neoplasms ; surgery ; Defecation ; Female ; Humans ; Intestinal Obstruction ; etiology ; surgery ; Male ; Middle Aged ; Proctocolectomy, Restorative ; Retrospective Studies

Since the outbreak of novel coronavirus pneumonia, the World Health Organization and the National Health Commission have issued multiple guidance documents.Medical facilities across the nation have actively implemented the countermeasures in response to the epidemic.However, in the real scenario of prevention and control, hospital infection remains a great concern.It is crucial to formulate measures and procedures based on standard prevention and transmission route prevention to avoid cross infection between medical workers and patients.Based on the practice, we summarize the institutional management, layout process, prevention and control of hospital infection in the emergency wards for further discussion and improvement.

OBJECTIVETo investigate the polymorphisms of 15 STR loci of Han population in Yan'an.

METHODSBlood samples were obtained from 100 unrelated Han individuals in Yan'an. DNA templates were screened by AmpF/STR Identifiler kit and ABI3100Avant DNA analyzer.

RESULTSThe allele frequencies of 15 STR loci ranged from 0.005 to 0.550, and the genotype frequencies ranged from 0.010 to 0.310. The combined match probability was 2.5x10(-17) and combined EPP was 0.999999999.

CONCLUSIONSThe 15 STR loci used in this study were highly polymorphic in Han population in Yan'an and suitable for population study and forensic cases in this region.

Alleles ; China ; ethnology ; Gene Frequency ; Genetic Markers ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

OBJECTIVETo investigate whether homocysteine (Hcy) participates the proliferation of the spontaneously hypertensive rat(SHR) vascular smooth muscle cell (VSMCs) and the molecular mechanism.

METHODSThe rat's arota were removed. The primary SHR VSMCs were isolated and cultured in vitro, then the SHR VSMCs were divided into four groups: (1) control group, (2) Hcy group, (3) 18alpha-glycyrrhetinic acid (GA) group, (4) Hcy + 18alpha-GA group. We detected proliferation of the SHR VSMCs by MTT and flow cytometry. The expression and co-localization of the connexin (Cx) 43 and Cx40 proteins in the SHR VSMCs were deteced by immunofluorescence. The expression of the Cx43 and Cx40 proteins in SHR VSMCs were detected by Western blot. The molecular dye transfer method (scrape dye transfer method) was applied to detect the gap junction function in the SHR VSMCs.

RESULTS(1) The Cx43 and Cx40 proteins expression in the SHR VSMCs were positive, confocal microscopy supported the co-localization of Cx43 and Cx40 in the cytoplasm. (2) The S value deteced by cell cycle and A value detected by MTT in the Hcy group were increased obviously compared with those in the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the S and A value in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (3) The expression of Cx43 and Cx40 proteins in Hcy group were increased compared with the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the expression of Cx43 and Cx40 proteins in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (4) The function of gap junction detected by scrape dye transfer method in the Hcy group were enhanced compared with the control group (P < 0.05), weakened in the 18alpha-GA group (P < 0.05). Compared with the Hcy group,the function of gap junction in the Hcy + 18alpha-GA group was significantly weakened (P < 0.05).

CONCLUSIONHcy can enhance the function of gap junctional to stimulate the proliferation of SHR VSMCs through the expression of Cx43 and Cx40 proteins promoted.

Animals ; Cell Proliferation ; Cells, Cultured ; Connexin 43 ; metabolism ; Connexins ; metabolism ; Gap Junctions ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; pharmacology ; Homocysteine ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Rats ; Rats, Inbred SHR