Objective To analyze the epidemiological and pathogenic characteristics and incidence trend of hand-foot-mouth disease (HFMD) in Jinshan District, Shanghai, and to provide data support for the prevention and control of HFMD． Methods Case information and etiological data of HFMD in Jinshan District from 2018 to 2023 were collected. Descriptive epidemiological methods were used to analyze the temporal, spatial and population distribution of HFMD cases and their etiological composition and changes. An autoregressive integrated moving average (ARIMA) model was established to predict the incidence trend of HFMD in 2024. Results From 2018 to 2023, a total of 5,979 cases of HFMD were reported in Jinshan District, with an average annual incidence rate of 123.00/100,000. There were no reports of severe cases or deaths. The incidence of HFMD showed unimodal distribution in 2018 and 2023, bimodal distribution in 2019, and there was no obvious peaks in 2020—2022. The town with the highest average annual incidence rate was Jinshanwei Town, and the town with the lowest average annual incidence rate was Fengjing Town. The male-to-female ratio of the cases was 1.42:1. Most of the cases were under 5 years old, and scattered children were the most common occupation. CVA6 was the predominant pathogen, but EV-A71 was not detected. The optimal fitting prediction model was SARIMAX (2, 0, 0) × (1, 0, 0, 12), and the model predicted a trend of decline after rising first in the incidence of HFMD in Jinshan District in 2024. Conclusion There are obvious temporal, spatial and population differences in HFMD incidence in Jinshan District, and the dominant pathogen of HFMD is CVA6. Prediction data can be used to further strengthen epidemic monitoring, timely detect new variants, and provide the basis for timely adjustment of prevention and control measures of HFMD.

Objective To investigate the relationship of mindful attention awareness with sleep quality and occupational stress of soldiers.Methods A questionnaire survey was conducted on 1 100 soldiers by using the mindful attention awareness scale(MAAS),occupational stress scale(OSS),and Pittsburgh sleep quality index(PSQI).Results The PSQI of the soldiers was negatively correlated with mindful attention awareness(r=-0.446,P<0.01),and positively correlated with occupational stress(r=0.569,P<0.01).Mindful attention awareness and occupational stress were effective predictors for sleep quality,accounting for 34.7%of variance.And the mediating effect of occupational stress between mindful attention awareness and sleep quality accounted for 58.86%.Conclusion The mindful attention awareness in soldiers can not only directly affect the sleep quality,but also indirectly affect the sleep quality through their occupational stress.

Objective:To assess and compare the performance of limiting-antigen avidity enzyme immunoassay (LAg-Avidity EIA) and pooling PCR in the surveillance for recent infection rates of HIV-1 in men who have sex with men (MSM).Methods:Blood samples were collected from MSM selected through snowball sampling method in sentinel surveillance in 13 prefectures of Yunnan province from 2016 to 2017. The samples were tested for HIV-1 antibody. The confirmed positive samples were tested by LAg-Avidity EIA. The negative samples were tested by pooling PCR. The recent infection rates of HIV-1 were estimated by the algorithm based on LAg-Avidity EIA and pooling PCR respectively. The two results were compared.Results:During 2016-2017, a total of 5 363 blood samples were collected from MSM, in which 407 samples were HIV-1 positive (including 177 positive tested previously) and 4 956 samples were HIV-1 negative. A total of 211 samples(91.7%) were tested by LAg-Avidity EIA, 69 were confirmed to be recent infections. A total of 4 469 samples were tested by pooling PCR, 8 were confirmed to be acute infections. The recent infection rates of HIV-1 from 2016 to 2017 estimated by LAg-Avidity EIA were 3.36% and 4.84%, and the recent infection rates estimated by pooling PCR were 3.27% and 3.02% respectively. The differences in recent infection rates of HIV-1 estimated by the two algorithms were not significant.Conclusions:The recent infection rates of HIV-1 estimated by LAg-Avidity EIA and pooling PCR in sentinel surveillance in MSM in Yunnan had good consistency from 2016 to 2017. Using the two methods might have a better stability in continuous surveillance for recent infection rates of HIV-1.

Objective:To understand the infection status of HCV and Treponema pallidum (TP) in HIV/AIDS cases in Yunnan province,and identify the risk factors. Methods:Between January 1 and June 30 in 2020,a cross-sectional survey was conducted in Yunnan. Two enzyme-linked immunosorbent assay (ELISA) kits were used to detect anti-HCV, the positive results of both two kits indicated HCV infection. ELISA and syphilis toluidine red untreated serum test were applied to identify TP infection. Both Excel 2016 and SPSS 22.0 software were used for statistical analysis, and logistic regression model was conducted to identify the relevant factors of HCV and TP infection.Results:A total of 5 922 HIV/AIDS cases were included in this study, the infection rates of HCV and TP were 6.5% (383/5 922) and 5.8% (344/5 922) respectively. The co-infection rate of HCV and TP was 0.4% (22/5 922). The risk for HCV infection in HIV/AIDS cases was higher in younger age groups compared with age group ≥50 years (15-19:a OR=3.53;20-29:a OR=3.02;30-39:a OR=2.91;40-49:a OR=3.61), in males than in females (a OR=2.31), in the married and unmarried than in the divorced or widowed (married:a OR=1.61;unmarried:a OR=1.63), in other ethnic groups than in Han ethnic group (a OR=1.70), in people with lower education level than in people with education level of college and above (primary school degree and below:a OR=4.69;middle school:a OR=3.96), in people living in the central and western Yunnan than in people living in eastern Yunnan (central Yunnan:a OR=2.46; western Yunnan:a OR=7.08), in injection drug users than in MSM (a OR=131.08). The risk of TP infection in HIV/AIDS cases was higher in people with education level of college and primary school than in middle school degree (primary school and below:a OR=1.73;college and above:a OR=1.77), in people with other occupations than in farmers (a OR=1.39), in people living in eastern Yunnan than in people living in western Yunnan (a OR=1.75); in MSM than in people with heterosex (a OR=9.75). Conclusions:A certain proportion of HIV/AIDS cases reported between January and June in 2020 in Yunnan were co-infected with HCV and TP, many factors were associated with the co-infection. It is suggested to strengthen HCV and TP tests in HIV/AIDS cases and conduct active treatment of the co-infection.

The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 mug/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicity in vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8 μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in alginate gel were killed by epirubicin.But few cells survived and some single-cell cloning spheres formed.Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation.Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo.The results showed that some cells positive for Oct3/4(TRITC)and Nanog(TRITC)were found in single-cell cloning spheres,and most of positive cells were concentrated in the core of sphere.Cells from spheres could form osteosarcoma in the body of mice.It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes(Oct3/4 and Nanog),resisting anti-cancer drugs,and tumorigenicity in vivo.To sum up,it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

BACKGROUND:The pathogenesis of intervertebral disc protrusion remains unclear.A stable and feasible intervertebral disc degeneration model is required to validate the pathology and imaging performance and to reveal intemal factors.OBJECTIVE:To investigate the feasibility of building rabbit model of intervertebral disc degeneration using aspiration method,as well as the convenience and stability of model establishment through the performance of pathology and imaging.MATERlALS:A total of 4210-month-old healthy New Zealand rabbits,irrespective of genders,were randomly divided into control group and experiment group with 21 rabbits in each group.Sumianxin was offered by Veterinary Institute,Quartermaster University of Chinese PLA in Changchun City.METHODS:The disc height index(DHI)of all rabbit swas set as 100%.A21-gauge hypodermic needle was used to aspirate nucleus pulposus 8-12 mg from L4-5 disc of rabbits in the experiment group,while control group received no management. At 4,12.20 weeks,7 rabbits were selected from each group for index determination.MAIN OUTCOME MEASURES:DHI percent,X-ray plain film,MRI and Alcian blue staining were used to observe the changes in the intervertebral disc degeneration.RESULTS:The DHI percentage of experiment group at 4,12 and 20 weeks respectively was(81.0±3.2)%,(75.0±2.5)%and (71.0±1.8)%,with significant difference compared to control group(P＜0.05).T2-weighted image showed that the signal of the discs was significantly decreased,and Alcian blue staining showed that the content of aggrecan was also significantly decreased.CONCLUSI0N:The aspiration is a reliable and convenient way to construct rabbit models of intervertebral disc degeneration.and these models exhibit the similar pathology and imaging performance with human intervertebral disc degeneration.

BACKGROUND: Pathological changes of pulmonary emphysema are not reversible according to the existent pathogenesis of pulmonary emphysema. Research over many years report that injury of pulmonary blood capillary may take part in new pathogenesis of pulmonary emphysema based on lung volume reduction operation and bronchial lumen occlusion. Mesenchymal stem cells (MSCs) have multi-directional differentiation potencies, such as the differentiation into vascular endothelial cells. Therefore, MSCs may promote pulmonary vascularization and repair pulmonary tissue.OBJECTIVE: To observe the effect of MSCs transplantation on pathological changes of arterial blood gas and pulmonary tissue in model rats with pulmonary emphysema, and investigate the therapeutic effects on MSCs on pulmonary emphysema and the pathogenesis of pulmonary emphysema.DESIGN: Randomized controlled animal study.SETTING: The Second Hospital of Shanxi Medical University.MATERIALS: Thirty healthy Wistar rats, 6 weeks old, of either gender, weighing 180-200 g. They were provided by Physiological Experiment Animal Center, Shanxi Medical University. All rats were randomly divided into MSCs treatment group, model group and control group with 10 rats each.METHODS: The experiment was carried out in the Physiological Laboratory of Shanxi Medical University from April 2005 to April 2006. Rats in the MSCs treatment group and in the model group were anesthetized and intratracheally perfused with 250 U/kg Porcine pancreatic elastase (PPE) to establish pulmonary emphysema models; while, rats in the control group were perfused with saline. The models were successfully established 4 weeks later. All rats were anesthetized and then femur and tibia were obtained to separate and culture MSCs in vitro. Immunocytochemistry was used to detect the expression of CD71 in order to evaluate MSCs. Bromium azacytidine-labeled MSCs were inserted along caudal vein into rats in the MSCs treatment group; while, rats in the model group and control group were inserted with the same volume of PBS solution.MAIN OUTCOME MEASURES: ① Changes of arterial blood gas in the three groups; ② Pulmonary tissue was used for pathological sections in order to calculate mean alveolar number, mean alveolar area and mean linear intercept; ③Immunocytochemical staining was used to measure numbers of CD34+ cells so as to determine proliferation of alveolar blood capillary.RESULTS: Three rats in all died during the model establishment, while another 3 rats were supplied. Therefore, an overall number of 30 rats were involved in the final analysis. ① Culture and evaluation of MSCs: At 3 days after inoculation, MSCs were generally adherent to walls and fusiformly shaped. In the third generation, the expression of CD71 was observed on the surface of MSCs.② Comparisons of arterial blood gas in the three groups: There were no significant differences in pH value, PO2, PCO2 and SaO2 in the three groups (P ＞ 0.05). ③ Pathological changes of pulmonary tissue: Pathological changes in the MSCs treatment group were milder than those in the model group;meanwhile, mean alveolar number in the MSCs treatment group was more than that in the model group, and there was significant difference between them (F=80.201, P＜ 0.05). While mean alveolar area and mean linear intercept in the MSCs treatment group were smaller than those in the model group, and there were significant differences (F =26.755,26.875, P ＜ 0.05). ④ Comparisons of CD34+ expression in pulmonary tissue: Relative positive area of CD34+ in the MSCs treatment group and model group was smaller than that in the control group (F =20.411, P ＜ 0.05), but that in the MSCs treatment group was larger than that in the model group, and there was significant difference between them (F=20.411, P＜ 0.05).CONCLUSION: MSCs can reverse the pathological changes of pulmonary emphysema; on the other hand, the decrease of the number of pulmonary capillary maybe one of the important pathogeneses of pulmonary emphysema.

VEGF nanoparticle (VEGF-NP) was prepared by a multi-emulsification technique using a biodegradable poly-dl-lactic-co-glycolic (PLGA) as matrix material. The nanoparticles were characterized for size, VEGF loading capacity, and in vitro release. VEGF-NP and naked VEGF plasmid were intramuscularly injected into the ischemia site of the rabbit chronic hindlimb ischemia model and the efficiency of VEGF-NP as gene delivery carrier for gene therapy in animal model was evaluated. Gene therapuetic effect was assessed evaluated by RT-PCR, immunohistochemistry and angiography assay. The average size of VEGF-NP was around 300 nm. The encapsulation efficiency of VEGF was above 96%. Loading amount of VEGF in the nanoparticles was about 4%. In vitro, nanoparticles maintained sustained-release of VEGF for two weeks. Two weeks post gene injection the capillary density in VEGF-NP group (81.22 per mm2) was significantly higher than that in control group (29.54 mm2). RT-PCR results showed greatly higher VEGF expression in VEGF-NP group (31.79au * mm) than that in naked VEGF group (9.15 au * mm). As a carrier system for gene therapy in animal model, VEGF-NP is much better than naked DNA plasmid. The results demonstrate great possibility of using NP carrier in human gene therapy.
Animals ; Disease Models, Animal ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; chemistry ; Lactic Acid ; chemistry ; Nanoparticles ; chemistry ; Plasmids ; Polyglycolic Acid ; chemistry ; Rabbits ; Vascular Endothelial Growth Factor A ; genetics

Tissue factor pathway inhibitor (TFPI) is one of the major physiological inhibitors of the human blood coagulation cascade and may have great potential in the prevention and therapy of diseases caused by thrombus formation. In this study, recombinant human tissue factor was generated in E. coli containing a recombinant vector constructed by inserting TFPI cDNA into pGEX-2T vector. The generated recombinant TFPI (rTFPI) could be simply purified with glutathione-agarose affinity method and maintained its biological function in terms of inhibition of tissue factor and factor Xa.
Escherichia coli ; genetics ; metabolism ; Humans ; Lipoproteins ; biosynthesis ; genetics ; pharmacology ; Peptide Fragments ; biosynthesis ; genetics ; pharmacology ; Plasmids ; Recombinant Proteins ; biosynthesis ; pharmacology ; Transfection