Sleep status and influencing factors among higher vocational college students / 四川精神卫生

BackgroundSleep disorder can adversely affect human physical and mental health， with important implications for socioeconomic development. And higher vocational college students are subject to various pressures and stimuli exerted by academic studies， job seeking and family socioeconomic status， which may contribute to the onset insomnia symptom or sleep disorder. ObjectiveTo discuss the sleep status and influencing factors among higher vocational college students， and to provide references for improving their sleep status. MethodsIn January and February， 2022， a stratified random sampling method was utilized to enroll 3 300 students from five higher vocational colleges in Wenjiang district of Chengdu city. All the participants were assessed using Insomnia Severity Index （ISI）， Patient Health Questionnaire Depression Scale-9 item （PHQ-9） and Generalized Anxiety Disorder Scale-7 （GAD-7）. Pearson correlation analysis was conducted to test the correlation of ISI with PHQ-9 and GAD-7 scores， and the Logistic regression analysis was performed to identify the influencing factors of insomnia symptom in higher vocational college students. ResultInsomnia symptom was detected in 81.90% （2 497 / 3 300） of higher vocational college students. The detection rate of insomnia symptom among vocational college students yielded statistical difference in terms of gender， family socioeconomic status， being an only child or having siblings， degree of psychological pressure exerted by academic studies or job seeking， amount of hours spent on Internet per day， frequency of physical exercise， and the presence or absence of anxiety symptom and depression symptom （χ2=21.032， 22.172， 8.983， 75.939， 36.781， 32.350， 54.512， 86.561， P<0.01 or 0.05）. Among higher vocational college students， ISI score was positively correlated with GAD-7 and PHQ-9 scores （r=0.620， 0.714， P<0.01）， and GAD-7 score was positively correlated with PHQ-9 score （r=0.824， P<0.01）. Gender， family socioeconomic status， degree of psychological pressure exerted by academic studies or job seeking， amount of hours spent on Internet per day， frequency of physical exercise， and the presence or absence of anxiety symptom and depression symptom were effective factors in predicting the insomnia symptom in higher vocational college students （P<0.01 or 0.05）. ConclusionFemale gender， average and below average family socioeconomic status， moderate-to-severe degree of psychological pressure exerted by academic studies or job seeking， spending 2~5 h， 5~7 h or >7 h on Internet per day， inadequate physical exercise and presence of depressive symptom are the risk factors for insomnia symptom in higher vocational college students.［Funded by Sichuan Provincial Primary Health Development Research Center in 2022， North Sichuan Medical College （number， SWFZ22-C-89）］

Changes of Th17/Treg proportion and the expressions of related cytokines in peripheral blood of patients with Hashimoto's thyroiditis / 中华地方病学杂志

Objective:To observe the ratio of helper T cells 17 (Th17)/regulatory T cells (Treg) in peripheral blood of patients with Hashimoto's thyroiditis (HT) and the expression changes of related cytokines in serum, and to explore their role in the occurrence and development of HT.Methods:Using the case-control study method, 35 HT patients examined in the General Hospital of Heilongjiang Beidahuang Group from February to November 2019 were selected as HT group, and 39 healthy people in the same period were selected as control group. Early morning fasting venous blood samples of the two groups were collected to test the levels of thyroid stimulating hormone (TSH), free thyroxine (FT 4), thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb). The expressions of serum interleukin (IL)-6, IL-17 and transforming growth factor β (TGF-β) were tested by enzyme-linked immunosorbent assay (ELISA); the number of Th17, Treg in peripheral blood were determined by flow cytometry. Results:The levels of TPOAb, TgAb and TSH in HT group [130.60 (43.37, 714.40), 368.10 (136.90, 1 103.00) U/ml, 9.05 (6.62, 15.23) μU/ml] were significantly higher than those in control group [2.66 (1.52, 4.69), 12.63 (11.43, 14.60) U/ml, 1.87 (1.36, 2.23) μU/ml, U = 6.87, 6.62, 4.85, P < 0.001], and the FT 4 level [0.76 (0.63, 1.04) ng/dl] was lower than that in control group [1.14 (1.02, 1.26) ng/dl, U = 7.39, P < 0.001]. The expressions of IL-6, IL-17 and TGF-β in HT group were higher than those in control group ( t = 2.41, 9.04, 2.44, P < 0.05). The number of Th17 and the ratio of Th17/Treg in HT group were higher than those in control group ( t = 4.20, 3.50, P < 0.05), and the number of Treg was lower than that in control group ( t = 4.45, P = 0.001). Conclusion:In HT patients, Th17 are increased, Treg decreased, Th17/Treg ratio increased, and the expressions of IL-6, IL-17 and TGF-β are increased.

Evaluation of miniprobe endoscopic ultrasonography for the diagnosis and treatment of rectal neuroendocrine neoplasms of different origin / 中华消化内镜杂志

Objective:To investigate the characteristics of endoscopic ultrasonography for rectal neuroendocrine neoplasms (R-NENs) of different origin and its influence on the diagnosis and treatment.Methods:A retrospective cross-sectional study was conducted to analyze 56 cases of R-NENs diagnosed by miniprobe endoscopic ultrasonography (MEUS) and/or pathology in the Endoscopy Center of Beijing Chaoyang Hospital, Capital Medical University from January 2016 to July 2021. The endoscopic ultrasonography characteristics, pathological features, surgical selection and the follow-up of R-NENs originating from deep mucosa and submucosa were compared.Results:Among the 56 patients, 49 were diagnosed as R-NENs.The diagnostic sensitivity, positive predictive value and diagnostic accuracy of MEUS for R-NENs were 93.88% (46/49), 86.79% (46/53) and 82.14% (46/56), respectively. R-NENs were mainly manifested by medium hypoechoic with MEUS [95.92% (47/49)]. Three R-NENs originated from submucosa were missed diagnosis, with 1 case presenting hypoechoic and 2 cases presenting hyperechoic. There were no significant differences in the tumor diameter, echo intensity under endoscopic ultrasonography, echo uniformity and pathological grade composition between deep mucosal origin and submucosal origin R-NENS (all P>0.05), but there was significant differences in the distance from tumor to anus ( χ2=5.011, P=0.025). The proportion of the distance from tumor to anus ≤5 cm of submucosal origin lesions was significantly higher than that of deep mucosal origin [43.75% (14/32) VS 17.65% (3/17)]. Endoscopic submucosal dissection [67.5% (27/40)] and transanal endoscopic microsurgery [25.0% (10/40)] were the major treatment method, but there were no significant differences in endoscopic ultrasonography manifestations and pathological grading of R-NENs between these two surgical procedures. Conclusion:There is no significant difference in endoscopic ultrasonography manifestations and pathological grade of R-NENs between deep mucosal origin and submucosal origin, suggesting that the prognosis is similar between the two types. It is no significant influence of endoscopic ultrasonography manifestations of R-NENs at different levels of origin.

Prevalence and influencing factors of mental health problems among hypertensive patients in Ya'an city five years after Lushan earthquake / 四川精神卫生

ObjectiveTo analyze the prevalence and influencing factors of mental health problems among hypertensive patients in Ya'an city five years after the Lushan earthquake， so as to provide references for promoting the mental health of hypertensive patients in earthquake-stricken areas. MethodsStratified random cluster sampling method was adopted to select 800 hypertensive patients in Ya'an city in December 2018. The general information of patients were collected via a self-designed questionnaire， and their mental health problems were assessed via the 12-item General Health Questionnaire （GHQ-12）.Then Logistic regression analysis was applied to explore the influencing factors. ResultA total of 744 valid questionnaires （93.00%） were obtained， and mental health problems were detected in 79 cases （10.62%）. Univariate analysis denoted that the detection rate of mental health problems yielded statistical difference among patients of different marital status （P<0.01）， family per capita monthly income （P=0.012） and social assistance recipients （χ2=25.194， P<0.01）. Logistic regression analysis indicated that the unmarried/separated/divorced/widowed status （OR=3.879， P=0.015） and social assistance recipients （OR=4.705， P<0.01） were risk factors for mental health problems among hypertensive patients. ConclusionThe detection rate of mental health problems is low among hypertensive patients in Ya'an city， while the unmarried/separated/divorced/widowed and social assistance recipients may suffer more serious mental health problems.

Effect of home visit based on empowerment education on psychotic symptoms， self-management ability and well-being of schizophrenia patients in home rehabilitation / 四川精神卫生

ObjectiveTo explore the effect of home visit based on empowerment education on psychotic symptoms， self-management ability and well-being of schizophrenia patients in home rehabilitation. MethodsA total of 87 cases of schizophrenia patients who met the diagnostic criteria of International Classification of Diseases， tenth edition （ICD-10） and were recovering at home in Caiba town， Yibin city from January to July 2021 were selected by random sampling method as research subjects. They were divided into experimental group （n=43） and control group （n=44） according to the random number table method. The two groups received routine family visits， and the experimental group received family visits based on empowerment education. The intervention cycle of the two groups was 6 months. Before and after the intervention， the severity of psychotic symptoms， self-management ability and well-being of the patients in the two groups were assessed with the Brief Psychiatric Rating Scale （BPRS）， the Schizophrenia Self-Management Instrument Scale （SSMIS） and the Index of Well-Being Scale （IWB）. ResultsAfter intervention， the intra group comparison showed that the BPRS score in the experimental group was lower than that before the intervention （t=4.550， P<0.01）， the SSMIS scores in both groups were higher than those before intervention （t=-17.107， -6.367， P<0.01）， and the IWB score in the experimental group was higher than that before intervention （t=-9.239， P<0.01）. The comparison between groups showed that the BPRS score of the experimental group was lower than that of the control group， and the SSMIS and IWB scores were higher than those of the control group （t=-3.899， 10.564， 9.690， P<0.01）. ConclusionThe implementation of family visit based on empowerment education may help to improve the psychotic symptoms of home rehabilitation schizophrenia， and improve their self-management ability and well-being.

Mental health status in close contacts of COVID-19 infections undergoing centralized quarantine and observation / 四川精神卫生

ObjectiveTo explore the prevalence rate and related risk factors of anxiety， depression and insomnia symptoms in close contacts of COVID-19 patients undergoing centralized quarantine and observation， so as to provide references for follow-up targeted psychological crisis intervention. MethodsFrom February 6 to 8， 2020， a total of 136 close contacts at designated quarantine sites in four districts and counties of Ya'an city were enrolled， and assessed using the general information questionnaire， Hospital Anxiety and Depression Scale （HAD） and Insomnia Severity Index （ISI）. ResultsA total of 26 quarantined individuals （19.12%） had various degrees of anxiety， depression or insomnia， including 16 cases （11.76%） with anxiety， 9 cases （6.62%） with depression and 16 cases （11.76%） with insomnia. Correlation analysis showed that HAD （A） score was positively correlated with HAD （D） score （r=0.642， P<0.01）. HAD （A） score was positively correlated with ISI score （r=0.590， P<0.01）. HAD （D） score was positively correlated with ISI score （r=0.329， P<0.01）. Multivariate Logistic regression analysis showed that the risk factor of anxiety was the first week of isolation （OR=3.290， P<0.05）， meantime， the risk factors of insomnia included the old age （OR=7.990， P<0.05） and the first week of isolation （OR=4.659， P<0.05）. ConclusionThe close contacts of COVID-19 infections undergoing centralized quarantine and observation in Ya’an have high prevalence rate of anxiety and insomnia， and the symptoms are most severe during the first week of quarantine.

Depressive symptoms detection among the urban elderly in Ya'an city and its influencing factors six years after Lushan earthquake / 四川精神卫生

ObjectiveTo investigate the prevalence and characteristics of depressive symptoms among urban elderly six years after Lushan earthquake in Ya'an， so as to provide references for the mental health interventions for elderly following catastrophic stressful life events. MethodsFrom March to April 2019， a multi-stage stratified cluster random sampling method was adopted to enroll 885 urban elderly people aged 60 and above in Ya'an. A self-compiled questionnaire was used to collect the general demographic information， health-related and disaster-related information， meantime， the elderly was assessed using Geriatric Depression Scale （GDS-30）. Thereafter， univariate and multivariate Logistic regression were applied to explore the influencing factors of depressive symptoms in urban elderly. ResultsA total of 783 valid questionnaires were collected， with a questionnaires response rate of 88.47%. Depressive symptoms were detected in 161 cases （20.56%）. The prevalence of depression showed statistical differences among the elderly of different gender， age， marital status， family relationship， monthly per capita household income， physical exercise status， health status， self-care ability， sleep status and disaster-affected degree （P<0.05 or 0.01）. Logistic regression analysis showed that the urban elderly of the female gender （OR=1.552， P=0.040）， monthly per capita household income of 2000~3000 yuan （OR=6.982， P<0.01）， monthly per capita household income≤2000 yuan （OR=6.857， P<0.01）， lack of physical exercise （OR=1.693， P<0.01）， being less capable of self-care （OR=3.838， P<0.01）， being incapable of self-care （OR=8.547， P<0.01）， complicating multiple curable diseases （OR=4.892， P<0.01） and complicating refractory chronic diseases （OR=5.657， P=0.031） were at high risk of depressive symptoms. The risk of depressive symptoms among the divorced or widowed elderly was greater than that among married elderly （OR=0.063， P<0.01）. ConclusionThe prevalence of depressive symptoms is relatively high among the urban elderly six years after Lushan earthquake in Ya'an， moreover， female gender， low monthly per capita household income， lack of physical exercise， being incapable of self-care and poor health status are risk factors affecting the depressive symptom， while being married is a protective factor.

The experimental research of dexamethasone reducing paclitaxel chemotherapy on lung cancer cells / 中国药理学通报

Aim: To investigate the effect of dexamethasone on paclitaxel chemotherapy, when given before chemotherapy. Methods: MTT method was used to investigate the effects of paclitaxel at different concentrations and dexamethasone (10 μ, mol · L-1) combined with paclitaxel at indicate dosages on the cell viability of lung cancer cells A549 and Calu-1. The cell cycle was detected by flow cytometry. The effect of dexamethasone on protein expression of glucocorticoid receptor (GR), P-glycoprotein (P-gP) and multidrug resistance protein-1 (MRP-1) expression were detected by Western blot. The effect of dexamethasone on lung cancer resistance of paclitaxel xenografts in nude mice was studied as well. Results paclitaxel at indicate dosages with dexamethasone (10 μmol · L-1) showed significantly increased IC50 value (A549: from 0.58 to 1.35 μmol · L-1; Calu-1 from 1. 10 to 2. 10 μmol · L-1). Dexamethasone activated GR, increased GR expression, and up-regulated the expression of P-gP and MRP-1. Dexamethasone combined with paclitaxel significantly decreased the antitumor effect of paclitaxel on nude mice transplanted tumor. Conclusions Giving dexamethasone before chemotherapy may cause resistance to cancer cells and reduce the effect of paclitaxel chemotherapy.

Treg/Th17 balance and immunology of schistosome infection: a review / 中国血吸虫病防治杂志

Many factors are reported to be involved in regulating the immunopathogenesis of schistosome infection. CD4+ T cell is one of the key players in the regulation of the liver granuloma formation by differentiation into different effector subsets including T helper (Th) 1, Th2, Th17, and T regulatory cells (Treg cells). Treg cells play an important suppressive role in immunopathology control and favor the pathogen to escape from the host immune assault. The functional activity of Tregs has been related to some autoimmune diseases including asthma and inflammatory bowel disease, which suggests that the manipulation of Tregs to restore their numbers and function may be therapeutic. However, interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. Therefore, a deeper understanding of the mechanisms of these immune regulations is necessary for the better control of pathology in schistosomiasis. In this paper, we review the Treg/Th17 balance and the immunology of schistosome infection.

Treg/Th17 balance and immunology of schistosome infection: a review / 中国血吸虫病防治杂志

Many factors are reported to be involved in regulating the immunopathogenesis of schistosome infection. CD4+ T cell is one of the key players in the regulation of the liver granuloma formation by differentiation into different effector subsets including T helper (Th) 1, Th2, Th17, and T regulatory cells (Treg cells). Treg cells play an important suppressive role in immunopathology control and favor the pathogen to escape from the host immune assault. The functional activity of Tregs has been related to some autoimmune diseases including asthma and inflammatory bowel disease, which suggests that the manipulation of Tregs to restore their numbers and function may be therapeutic. However, interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. Therefore, a deeper understanding of the mechanisms of these immune regulations is necessary for the better control of pathology in schistosomiasis. In this paper, we review the Treg/Th17 balance and the immunology of schistosome infection.

Optimal conditions for transfecting Wharton's Jelly-derived mesenchymal stem cells with electroporation method / 中国组织工程研究

BACKGROUND: Wharton's Jelly-derived mesenchymal stem cells are relatively primitive stem cells that are ideal vectors for gene therapy. However, there is a lack of studies on the conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells. Therefore, exploring the optimal conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells occupies an important position. OBJECTIVE: To investigate the effects of different electroporation conditions on the transfection efficiency of Wharton's Jelly-derived mesenchymal stem cells, and to explore the optimal conditions for cell electroporation. METHODS: By controlling the transfection conditions such as voltage, pulse duration and cell status, EEV-EGFP plasmids were transfected into Wharton's Jelly-derived mesenchymal stem cells by electroporation under different conditions. Transfection efficiency was detected by flow cytometry. RESULTS AND CONCLUSION: (1) The transfection efficiency was intended to increase when the voltage ranged from 125 V to 150 V, and the maximum transfection efficiency was obtained when the voltage was 150 V. However, when the voltage was further increased to 170 V, the transfection efficiency began to decrease considerably. (2) The maximum transfection efficiency was obtained when the pulse duration was 5.0 ms, while it was certainly decreased when the pulse duration was 2.5 and 7.5 ms. (3) The transfection efficiency of the cells cultured under normoxia was higher than that under hypoxic culture. These findings reveal that normally cultured Wharton's Jelly-derived mesenchymal stem cells can achieve higher electroporation efficiency via two pulse sessions at a voltage of 150 V, pulse duration of 5.0 ms, and pulse interval time of 50 ms.

Potential risk factors of excessive epistaxis after endoscopic endonasal surgery / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the potential risk factors and management of excessive epistaxis after endoscopic endonasal surgery (EES).@*METHOD@#Six hundred and forty-one patients who underwent EES in our hospital from December 2011 to December 2012 were reviewed retrospectively. Factors which potentially affect the incidence of excessive epistaxis after EES were analyzed with univariate and multivariate logistic regression model.@*RESULT@#The incidence rate of excessive epistaxis after EES was 8.4% in our study. Multivariate logistic regression analysis revealed that history of previous EES, along with other four factors, correlated significantly with the occurrence of excessive epistaxis after EES.@*CONCLUSIONS@#Previous EES, along with other three factors, may increase the chance of excessive epistaxis after EES, while pre-operative corticosteroid therapy may reduce the risk to some extent.

Influencing factors for health-related quality of life in patients with chronic liver disease / 中华全科医师杂志

Objective To investigate the factors affecting health-related quality of life (HRQOL) in patients with chronic liver disease (CLD).Methods HRQOL was measured with SF-36v2 Chinese version.All patients with CLD diagnosed between December 2009 and May 2011 in Liver Research Center,Beijing Friendship Hospital and Digestive Department,Beijing Chaoyang Hospital were enrolled in this study.Patients with CLD were divided into 4 groups according to their Child-Turcotte-Pugh scores:no cirrhosis,Child's classes A,B,and C.Demographic and clinical data were collected in each group.Results A total of 392 patients with CLD and 91 healthy controls were included.HRQOL in patients with CLD was lower than that in healthy controls.Physical component score (PCS) in healthy controls was 54.6 ± 5.5,and in CLD was 47.8 ± 8.8 (t =9.343,P ＜ 0.01).Mental component score (MCS) in healthy controls was 56.4 ± 8.1,and in CLD was 51.7 ± 7.4 (t =5.302,P ＜ 0.01).Increasing severity of CLD from no cirrhosis to advanced cirrhosis was associated with a decrease on HRQOL,PCS scores were 53.1 ± 5.9,48.7 ± 6.6,42.4 ± 7.1 and 37.2 ± 8.9 (F =93.353,P ＜ 0.01) ; MCS scores were 53.2 ± 4.8,52.7 ±6.5,51.8 ±7.5,and 46.8 ± 7.5 (F =11.325,P ＜0.01),for no patients with cirrhosis,Child's classes A,B and C respectively.Stepwise linear regression analysis showed that severity of disease,age,present ascites,present varices,and prothrombin time had significant effects on physical health area (F =100.893,P ＜ 0.010).Severity of disease,female gender,present varices,total bilirubin,prothrombin time and hemoglobin had significant effects on mental health area (F =19.237,P ＜ 0.010).Conclusions Patients with CLD have reduced HRQOL.Increasing severity of CLD is associated with a decreasing HRQOL.Old age,female gender,advanced stage of CLD,present ascites,hyperbilirubinemia and prolonging prothrombin time are risk factors for reducing HRQOL.

Diagnostic value of S100A6 mRNA expression level in endoscopic ultrasonography guided fine needle aspiration specimens in pancreatic cancer / 中华消化杂志

ObjectiveTo investigate the feasibitity of detecting S100A6 expression at mRNA level in endoscopic ultrasonography guided fine needle aspiration (EUS-FNA) pancreatic ductal adenocarcinoma (PDA) specimens and its diagnostic value in PDA.MethodsA total of 18 PDA specimens and 22 normal pancreatic specimens were collected. RNA was extracted for reverse transcription.The expression of S100A6 gene was examined by real-time polymerase chain reaction.The cut-off value of S100A6 expression at mRNA level in PDA diagnosis was established through receiver operating characteristic (ROC) analysis. 28 patients with pancreatic head masses were selected for EUS-FNA examination,and the value of S100A6 mRNA expression level in PDA diagnosis was prospectively evaluated. The expression of S100A6 protein in PDA tissue was determined by immunohistochemistry staining.ResultsS100A6 mRNA expression in EUS-FNA and surgical PDA specimens (0.05023±0.10120,0.02083 ± 0.02848) was significantly higher than that of normal pancreatic tissues (0.00164±0.00202),both P＜0.01.The expression of S100A6 in 22 EUS-FNA PDA specimens was significantly higher than that of 6 pancreatic benign disease biopsy specimens (0.00193 ± 0.00278,P =0.0009). There was no significant difference in S100A6 expression between 6 pancreatic benign disease biopsy specimens and normal surgical pancreatic samples (P=0.6143).When S100A6 mRNA expression in EUS-FNA specimens over 0.00525 was taken as positive diagnostic value,the sensitivity,specificity and accuracy in prospective pancreatic cancer diagnosis were 90.01％,100 ％ and 92.85 ％,respectively.ConclusionThe high expression of S100A6 mRNA in EUS-FNA specimens of PDA has good preoperative diagnostic value.

Clinical analysis of regional portal hypertension / 中华消化杂志

Objective To investigate the etiology, clinical features, diagnosis and treatment of regional portal hypertension (RPH).Methods The clinical data of 26 patients with RPH treated in Beijing Chaoyang Hospital Affiliated to Capital Medical University between January 2005 and June 2010 were analyzed with retrospective analysis.The first symptom, routine analysis of blood, liver function test, hepatitis B and C markers, tumor markers, abdominal ultrasound, abdominal enhanced CT, endoscopy findings of 26 patients and the results of abdominal CT angiography (CTA) of 16cases were analyzed.Results Pancreatic disease (18 cases) was the leading cause of RPH.The main clinical manifestations of splenomegaly in 26 cases, irregularly abdominal pain in 14 cases, and upper gastrointestinal bleeding in 10 cases.Isolated gastric varices were revealed by endoscopy in 25 cases,complicated with lower esophageal varices in 1 case.4 cases with endoscopic tissue glue injection in gastric variceal bleeding, splenectomy in 4 cases, 2 cases with splenectomy and pericardialdevascularization, 2 cases with splenectomy, pancreatic tail resection and spleno-renal shunt, 3 cases with splenic embolization treatment.Conclusions RPH often accompanied by pancreatic disease,manifested as splenomegaly, hypersplenism, but normal liver function, absence of liver cirrhosis.Isolated gastric varices is the characteristic features of RPH.RPH caused by benign diseases is curable.Splenectomy is more effective than simple endoscopic hemostasis in RPH associated with gastrointestinal bleeding.

Transfection of bone marrow mesenchymal stem cells by adeno-associated virus 2/1 vector / 中国实验血液学杂志

This study was aimed to investigate the transfection efficacy of recombinant adeno-associated virus 2/1 (rAAV2/1) on bone marrow mesenchymal stem cells (BMMSCs) at different multiplicities of infection (MOI) and time, and effect of transfection on growth of rat BMMSCs. The rat BMMSCs cultured in vitro were transfected by using rAAV2/1 with enhanced green fluorescent protein (rAAV2/1-EGFP) at MOI of 1 x 10(4), 1 x 10(5) and 1 x 10(6); the EGFP expression was observed by fluorescent microscopy at 3, 7 and 14 days. The viability, proliferation multiple, differentiation ability of daughter cells were detected for evaluating the effect of rAAV2/1 on survival, proliferation and differentiation of BMMSCs and the fluorescence index (FI) were determined by flow cytometry. The results indicated that after transfection with rAAV2/1 for 24 hours the green fluorescence in BMMSCs were observed, but also the fluorescence gradually was enhanced along with prolonging of time, and reached to steady level after 7 days; the viability, proliferation multiple, differentiation ability of BMMSCs transfected by rAAV2/1-EGFP at different MOI showed no significant changes at 3,7 and 14 days (p > 0.05), meanwhile at same MOI the proliferation multiple obviously increased in comparison between 7 day vs 3 day and 14 days vs 7 days (p < 0.01). The flow cytometric detection showed that the transfection efficacy of rAAV2/1-EGFP on BMMSCs and FI increased significantly as the multiplicity of infection and culture time increased (p < 0.05). It is concluded that rAAV2/1-EGFP is able to transfect into BMMSCs effectively, but the transfection efficiency and fluorescence index increase significantly along with increase of multiplicity of infection and culture time. rAAV2/1-EGFP do not affect viability, proliferation multiple and differentiation ability of BMMSCs. rAAV2/1 is a kind of active vector for gene transfer to reform BMMSCs.

Secretory expression of recombinant porcine zona pellucida glycoprotein-3alpha (rpZP3alpha) in Pichia pastoris / 生物工程学报

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.

The autologous bone marrow mononuclear cell transplantation by intracoronary route treat patients with severe heart failure after myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the chronic effects of intracoronary autologous bone marrow mononuclear cell (BM-MNCs) transplantation in patients with refractory heart failure (RIHF) after myocardial infarction.</p><p><b>METHODS</b>Thirty patients with RIHF (LVEF < 40%) were enrolled in this nonrandomized study, autologous BM-MNCs (5.0 +/- 0.7) x 10(7) were transplanted with via infarct-related coronary artery in 16 patients and 14 patients received standard medical therapy served as control. Baseline and follow up evaluations included complete clinical evaluations, plasma BNP, ANP, ET-1 measurements, echocardiography, PET, and Holter monitoring.</p><p><b>RESULTS</b>Baseline characteristics were similar between the 2 groups. There were no major periprocedural complications. One patient developed ventricular premature contractions during cell infusion for several seconds and recovered spontaneously. Compared to pre-transplantation, plasma BNP and ET-1 significantly decreased and plasma ANP significantly increased at 7 days post transplantation; 6 minutes walking distance increased from (72.1 +/- 31.5) to (201.6 +/- 23.3) m (P < 0.01), LVEF increased 9.9% (P < 0.001) and FDG-PET revealed vital myocardium area increased (10.3 +/- 3.4)% (P < 0.01) at 3 months after BM-MNCs transplantation. At 6 months follow up, the NYHA class improved from (3.4 +/- 0.1 to 2.4 +/- 0.2, P < 0.001) and no patient died and 1 patient rehospitalized due to lower extremities edema. In control group, LVEF decreased 7.2% compared to baseline (P < 0.001) and was significantly lower than transplantation group at 3 months (P < 0.001). At 6 months follow up, the NYHA class increased from (3.5 +/- 0.1 to 3.9 +/- 0.1, P < 0.05), 2 patients died and 10 patients rehospitalized due to aggravated heart failure.</p><p><b>CONCLUSION</b>Present study demonstrates that intracoronary transplantation of autologous BM-MNCs is safe and effective for treating patients with RIHF after myocardial infarction.</p>