The double-antibody-sandwich enzyme-linked immunoadsorbent assay (ELISA) for detection of rLysostaphin in humans had been developed and established through this study. rLysostaphin of high purity ( > 95 % ) produced in Shanghai Hi-Tech United Bio-Technological Research & Development Co., Ltd (SHUBRD) was used to produce a rabbit anti-rLysostaphin polyclonal antibody. The standard curve of rLysostaphin polyclonal antibody that was constructed showed that the lowest range of detection was found at 0. 98 ng of rLysostaphin/mL, and the curve exhibited linearity preferably from 0. 98 to 500 ng of rLysostaphin/mL. When three serum samples of the same batch were assayed for 6 replicates, and more 3 samples from different batches for 6 replicates, the average intra-assay and inter-assay coefficient variances ( CV) were 6. 4% and 6. 5%, respectively. The relative recovery rate was 98.6% when quantitative standard antigens were added to the serum. The present method for detection of rLysostaphin in serum is specific, highly sensitive, highly precise, and exhibited a low CV and will be helpful in the further study of rLysostaphin pharmacokinetics and holds promise in clinical applications.
Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Enzyme Stability ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immune Sera ; immunology ; Lysostaphin ; blood ; immunology ; metabolism ; Male ; Rabbits ; Recombinant Proteins ; immunology ; metabolism ; Reproducibility of Results ; Temperature

Objective: To observe the effects of herb-partitioned moxibustion and ginger-partitioned moxibustion on the growth of colon tumors in rats with colitis-associated colon cancer (CACC), and explore the mechanism of moxibustion intervening CACC through the purinergic receptor P2X ligand-gated ion channel 7 (P2X7R)/signal transducer and activator of transcription 3 (STAT3)/vascular endothelial growth factor (VEGF) pathway. Methods: A total of 26 male Sprague-Dawley rats were selected. According to the random number table method, 6 rats were selected as the normal group. The remaining 20 rats were injected intraperitoneally with azoxymethane (AOM) combined with oral dextran sodium sulfate (DSS) to prepare the CACC model. After the model was successfully established, 2 rats were randomly selected for model identification. The remaining 18 rats which were successfully modeled were randomly divided into a model group, a herb-partitioned moxibustion group and a ginger-partitioned moxibustion group, with 6 rats in each group. Moxibustion intervention was performed in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group at Qihai (CV 6) and bilateral Tianshu (ST 25). Moxibustion was performed twice at each point each time, once a day, at a 1-day interval after 6 consecutive interventions, for a total of 30 interventions. After intervention, the colon tumor load, pathological change and histopathological score were observed. Immunohistochemistry was used to detect the expressions of VEGF, P2X7R, phospho-STAT3 (p-STAT3), and nuclear factor-kappa B p65 (NF-κB p65) proteins in rat colon tissue. Western blot was used to detect the levels of p-STAT3 and NF-κB p65 proteins in rat colon tissue. Results: Compared with the normal group, the colon tumor load and histopathological score in the model group were significantly increased (both P<0.001), and different grades of dysplasia were observed in colon tissue from the model group, reaching the degree of adenocarcinoma; the expression level of P2X7R protein in colon tissue was significantly decreased (P<0.001), and the expression levels of p-STAT3, NF-κB p65 and VEGF proteins were significantly increased (all P<0.001) in the model group. Compared with the model group, the colon tumor load, colon histopathological score and the levels of p-STAT3, NF-κB p65 and VEGF proteins in colon tissue were significantly decreased (all P<0.05) in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group while the expression levels of P2X7R protein in colon tissue were significantly increased (both P<0.05). Conclusion: Both herb-partitioned moxibustion and ginger-partitioned moxibustion can reduce the colon tumor load in CACC rats and delay the progression of colon adenomas. The mechanism may be mediated by the P2X7R/STAT3 pathway to inhibit STAT3 phosphorylation, thereby reducing VEGF protein expression.

Objective To understand the dietary intake of selenium and its influencing factors among rural and urban residents in Zhejiang province. Methods From 2010 to 2012,a total of 2,659 residents were selected from large urban sites,small-medium urban sites and rural sites in Zhejiang. Using 24 -hour dietary recall method,a 3 -day household dietary survey to analysis the dietary intake of selenium. Results The average daily dietary intake of selenium for residents aged 1-,4-,7 -,11 -,14 -,18 -,45 - and 60 - was 21. 96,26. 39,31. 62,35. 26,29. 39,41. 78,39. 12 and 38. 40 μg,respectively. According to formulation of Chinese Nutrition Society,the dietary selenium intake of 42. 56%juveniles and 52. 09% of adults was below the estimated average requirement( EAR). Significant statistical differences were found between normal group and insufficient dietary selenium intake group in terms of age,sex,region,level of education, per capita annual income,physical exercise and smoking status(all P<0. 01). Influencing factors of insufficient selenium intake were female(OR =1. 86,95%CI:1. 59 -2. 63),rural area( OR =1. 46,95%CI:1. 23 -1. 73),lower level of education(OR=0. 70,95%CI:0. 57 -0. 86)and lower income( OR =0. 72,95% CI:0. 60 -0. 88). Conclusion Influencing factors of dietary selenium intake are sex,region,level of education and per capita income. Dietary selenium supplement should be strengthened through a variety of ways,especially in target population.

Objective TolearntheproteinintakestatusofadultsinZhejiangProvinceandtoinvestigatetherelationship between the protein intake and influencing factors,and in order to provide a scientific basis for improving adult protein intake.Methods Datawereselectedfromthe2010—2012ChineseNationalNutritionandHealthSurveyinZhejiang province.Data were gained through medical examination and the method of 3 day 24-hour dietary recall and food weighted record.Descriptiveanalysiswasconducted.Results Therewere1160men(48.09%)and1252women(51.91%)in 2 412 cases in this analysis.The average protein intake per day was 71.87 g for per reference man,and the intake quartile was 66.06(51.17-85.93)g/d and 38.14% of adults were not achieved the Recommended Nutrient Intake (RNI).The multi factor logistic regression analysis showed that age,region,income and occupation were the main factors affecting protein intake.Age was a risk factor,while income was a protective factor.Big cities were more easily to have insufficient protein intake than median and small cities.Students,agriculture,housework and other groups of adults were more easily to have insufficient protein intake.And 30.70% of dietary protein was from cereal,and 39.70% was from animal food. Sources of dietary protein were statistical significant among different areas and age.Protein intake from cereal among young people (29.60%),old people (29.95%),and people living in big cities (19.81%)was low.Protein intake from cereal among people living in medium and small cities (10.40%)was high.Young people (43.12%)and people living in big cities(52.87%)hadhigheranimalsourceprotein.Conclusion TheproteinintakeofadultsinZhejiangProvincewasnot achieved the RNI.It is important to conduct health educations according to the protein intake problems of different groups to improve their protein intake status.

OBJECTIVETo understand the immunological status of Japanese encephalitis (JE) antibodies amongst migrant workers and to provide epidemiological basis for public health strategies on JE prevention and control in Shenzhen.

METHODSA multi-stage random sampling method was used, and 1003 migrant workers aged 18 to 60 from 44 factories were investigated and their serum specimens were collected. The enzyme-linked immunosorbent assay (ELISA) was used to detect JE antibodies qualitatively.

RESULTSThe gross IgG seroprevalence rate for JE was 20.2% (203/1003). Sex-specified seroprevalence was 21.2% (103/485) for male and 19.3% (100/518) for female, respectively (χ(2) = 579, P > 0.05). Age-specific seropositive rates were 22.6% (12/53) for those below 20 years old, 18.7% (120/642) for those between 20-years old, 26.0% (58/223) for those between 30-years old and 15.3% (13/85) for those on or above 40 years old (χ(2) = 7.96, P > 0.05). Proportions for self-reported positive immunization, non-immunization and unclear immunization history were 22.1% (30/136), 22.1% (51/231) and 19.2% (122/636), respectively (χ(2) = 501, P > 0.05). Seroprevalence by region of origins showed that workers from Guangdong province was the highest (30.5%, 50/164), followed by workers from Guangxi (29.7%, 22/74) whilst workers from Shan(3)xi (5.4%, 2/37) had the lowest rate. Seroprevalence rate for managers (29.0%, 31/107) was higher than that of technicians (7.1%, 1/14) (χ(2) = 21.78, P < 0.05). Serological positive rate of workers with university or above educational background was the highest (32.7%, 16/49), followed by that for individuals with college degree (10.3%, 10/97) (χ(2) = 13.02, P < 0.05).

CONCLUSIONNo associations are detected between JE seroprevalence and age, or sex, or self-reported immunization histories amongst migrant labor workers in Shenzhen. However, correlations between JE serological positive rate and region of origins, occupation and educational attainment are found to be significant. The gross seroprevalence of JE antibodies suggests that the level of JE antibodies amongst Shenzhen migrant workers is low and the population immunity barrier has yet to be established. It is necessary to strengthen prevention and control strategies of JE among labor workers of Shenzhen.

Adolescent ; Adult ; Antibodies, Viral ; blood ; China ; epidemiology ; Encephalitis, Japanese ; epidemiology ; prevention & control ; Female ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; Male ; Middle Aged ; Transients and Migrants ; statistics & numerical data ; Young Adult

OBJECTIVETo investigate the effects of heparanase expression inhibition on the proliferation, invasiveness and apoptosis of human lung adenocarcinoma cell line A549 cells.

METHODSRecombinant eukaryotic expression plasmid pshRNA-Hpa targeting human heparanase gene was constructed. A549 cells were cultured in DMEM and transfected with pshRNA-Hpa. The expression of heparanase mRNA and protein were examined by RT-PCR and Western blot. The proliferation, invasiveness and apoptotic rates of A549 cells were determined by MTT method, matrigel invasion assays and flow cytometry respectively.

RESULTSThe expression levels of heparanase mRNA and protein were down-regulated in A549 transfected with pshRNA-Hpa. The number of cells penetrating matrigel and the proliferation ability of A549 cells transfected with pshRNA-Hpa were reduced significantly compared to the control cells. The apoptotic rate of A549 cells transfected with pshRNA-Hpa was 12.53% +/- 0.34%, being significantly higher than that of the control cells (both P < 0.01). Western-blot showed that inhibition of heparanase expression led to reduced Akt phosphorylation.

CONCLUSIONSThe recombinant plasmid pshRNA-Hpa effectively inhibited the expression of heparanase, thus suppressing the proliferation and invasion and inducing apoptosis of A549 cells. The effects may be due to the down-regulation of Akt phosphorylation level.

Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Glucuronidase ; antagonists & inhibitors ; metabolism ; Humans ; Lung Neoplasms ; enzymology ; pathology ; RNA Interference ; immunology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVETo evaluate long-term outcomes of minor liver resection for hilar cholangiocarcinoma (HC) of Bismuth-Corlette type III.

METHODSFrom January 1997 to December 2007, the clinical data of 91 patients with Bismuth-Corlette type III HC underwent hepatectomy were collected and analyzed retrospectively.

RESULTSThere were 60 patients underwent minor hepatectomy, and 31 undergoing major hepatectomy. Hepaticojejunostomy was made conventionally in an end-to-side fashion in the patients undergoing major liver resection, and a new technique of hepaticojejunostomy used in the patients undergoing minor liver resection. That was the anterior edges of bile duct stumps which were not sutured after suturing of posterior edges. Instead of, the anterior edge of jejunum loop to the remnant liver on the top of the bile duct stumps were sutured with intermittent "U" sutures. In all patients, in-hospital mortality rate was 0 and rate of bile leakage was only 2.1%. The actual 1-, 3- and 5-year survival rates were 91.6% and 87.0%, 61.6% and 62.0%, 31.6% and 33.0%, respectively (P > 0.05).

CONCLUSIONSMinor liver resection for the selected patients with HC of Bismuth-Corlette type III according to our criteria achieved better long-term outcomes. A new hepaticojejunostomy used in the patients undergoing minor liver resection is a safe and effective method.

Adult ; Aged ; Bile Duct Neoplasms ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; surgery ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

Adult ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; prevention & control ; Male ; Middle Aged ; Vaccination

This study was aimed to assess the effect of Astragalus Polysaccharide (ASPS) on in-vitro hematopoiesis. CFU-GM assays were used to determine the effect of ASPS and thrombopoietin (TPO) on granulocytic-monocyte progenitor cells. The CFU assays were also used to investigate the effect of ASPS on the proliferation of HL-60 cells.HL-60 cells were cultured with serum-free RPMI 1640 medium and treated with or without of different concentrations of ASPS. After 72 h incubation, the number of cells were counted.In addition, the caspase-3 and JC-1 expression was determined by flow cytometry with Annexin V/PI double staining. The results showed that ASPS (100, 200 µg/ml) and TPO (100 ng/ml) significantly promoted CFU-GM formation in vitro. Various concentrations of ASPS and TPO also promoted the colony formation of HL-60 cells, the largest effect of ASPS was observed at a concentration of 100 µg/ml. There were no synergistic effects between TPO and ASPS on cellular proliferation. The results also showed that ASPS significantly protected HL-60 cells from apoptosis in condition of serum-free medium culture, suppressed caspase 3 activation, and reduced the cell apoptosis. It is concluded that ASPS can significantly promote the formation of bone marrow CFU-GM and the proliferation of HL-60 cells, the optimal concentration of ASPS is at 100 µg/ml. In the absence of serum inducing apoptosis, ASPS also significantly reduced the apoptosis of HL-60 cells via suppressing the activation of caspase-3.
Apoptosis ; drug effects ; Astragalus Plant ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; HL-60 Cells ; Hematopoiesis ; drug effects ; Humans ; Polysaccharides ; pharmacology ; Thrombopoietin ; pharmacology

Background: Sleep disorders are one of the earliest non-motor symptoms of Parkinson’s disease (PD). Sleep disorders could, therefore, have value for recognition and diagnosis in PD. However, no unified classification and diagnostic criteria exist to evaluate sleep disorders by polysomnography (PSG). Utilizing PSG to monitor sleep processes of patients with PD and analyze sleep disorder characteristics and their relationship with demographic parameters could aid in bridging this gap. This preliminary study aimed to evaluate the clinical characteristic of sleep disorders in PD using PSG. Methods: PSG was used to evaluate sleep disorders in 27 patients with PD and 20 healthy volunteers between August 2015 and July 2018 in Fujian Medical University Union Hospital. Total sleep time (TST), sleep efficiency (SE), total wake time, and other parameters were compared between the two groups. Finally, the correlation between sleep disorders and age, disease duration, Unified Parkinson’s Disease Rating Scale-III scores, Hoehn-Yahr stage, and levodopa dose were analyzed. The main statistical methods included Chi-square test, two independent samples t test, Fisher exact test, and Pearson correlation. Results: Sleep fragmentation in the PD group was significantly increased (74.1%) while difficulty falling asleep and early awakening were not, as compared to healthy controls. No significant differences were found in time in bed, sleep latency (SL), non-rapid eye movement (NREM) stage 1 (N1), N1%, N2, N2%, N3%, and NREM% between PD and control groups; but TST (327.96 ± 105.26 min vs. 414.67 ± 78.31 min, P = 0.003), SE (63.26% ± 14.83% vs. 76.8% ± 11.57%, P = 0.001), R N3 (20.00 [39.00] min vs. 61.50 [48.87] min, P = 0.001), NREM (262.59 ± 91.20 min vs. 337.17 ± 63.47 min, P = 0.003), rapid-eyemovement (REM) (32.50 [33.00] min vs. 85.25 [32.12] min, P < 0.001), REM% (9.56 ± 6.01 vs. 15.50 ± 4.81, P = 0.001), REM sleep latency (157.89 ± 99.04 min vs. 103.47 ± 71.70 min, P = 0.034) were significantly reduced in PD group. Conclusion This preliminary study supported that sleep fragmentation was an important clinical characteristic of sleep disorders in PD. Whether sleep fragmentation is a potential quantifiable marker in PD needs to be further investigated in the future study.