OBJECTIVE To synthesize a polymer PEI-ss-PEG2000-DSPE containing disulfide bond and to prepare as cationic micelle(P-ss-PD) based on branched polyethyleneimine(PEI). To investigate the ability of P-ss-PD micelle to reduce cytotoxicity and improve the transfection efficiency of antisense oligonucleotide(ASO) in human breast cancer cell lines, and to study the anti-tumor effect of P-ss-PD micelle in nude mice. METHODS PEI-ss-PEG2000-DSPE was synthesized by grafting PEG2000-DSPE onto branched PEI with disulfide bond as a connecting arm. P-ss-PD micelle was prepared by ethanol injection method and P-ss-PD/ASO nanocomplex was obtained by combining P-ss-PD micelle with ASO. The particle size and zeta potential of P-ss-PD/ASO nanocomplex at various mass ratios were determined by laser particle size analyzer. Agarose gel retardation assay was used to investigate the binding degree of P-ss-PD/ASO nanocomplex and determine the optimal mass ratio. At the same time, the reduction responsive ability of P-ss-PD micelle was investigated. The cytotoxicity of P-ss-PD micelle was detected by CCK8 kit. The transfection efficiency of P-ss-PD micelle was investigated by flow cytometry and high content cell imaging analysis system in MDA-MB-231 cells. The anti-tumor effect of P-ss-PD micelle was investigated by tumor-bearing nude mice models. RESULTS When the mass ratio was 300∶1, the particle size of P-ss-PD/ASO nanocomplex was the smallest and had a good stability. The average particle size was (58.90 ± 4.08)nm, the average zeta potential was (16.80 ± 1.23)mV, and the morphology was uniform spherical. P-ss-PD/ASO nanocomplex had the reduction responsive ability and could release ASO under highly reductive conditions.In vitro, compared with unmodified branched PEI, the cytotoxicity of P-ss-PD micelle was significantly reduced and the transfection efficiency was significantly increased.In vivo, the tumor growth inhibition rate of P-ss-PD/ASO nanocomplex in tumor-bearing nude mice was more than 50%. CONCLUSION The P-ss-PD micelle prepared in this study is a kind of low toxicity and high transfection efficiency non-viral vector, which has the characteristics of reduction responsive releasing, and shows a promising application in ASO drug delivery.

Objective:To evaluate and summarize the best evidence for exercise scheme after bariatric and metabolic surgery in obese patients at home and abroad.Methods:We used "obesity" " bariatric and metabolic surgery" "sleeve gastrectomy" "gastric bypass surgery" "banding surgery" "exercise" "physical activity" "exercise therapy" "guideline" "expert consensus" "systematic review" "Meta-analysis" and "evidence summary" as keywords, and searched databases such as UpToDate, Cochrane Library, PubMed, China National Knowledge Infrastructure (CNKI) , Wanfang Data, and the guide network and professional association platforms by computer. Literature retrieved included guidelines, expert consensus, best clinical practice, clinical decision-making, systematic reviews, and the search time limit was from the database establishment to July 18, 2020. Two researchers evaluated the quality of literature and extracted evidence, and the evidence was revised and perfected through expert consultation.Results:A total of 4 systematic reviews and Meta-analysis articles, 2 clinical decision-making articles, 4 guidelines, and 1 expert consensus were included. The evidence was collected from 7 aspects, namely the importance of exercise, pre-exercise evaluation, exercise style, exercise intensity and duration, exercise safety, exercise follow-up, exercise effect evaluation, and finally 33 best evidences were summed up.Conclusions:Evidence of exercise scheme after bariatric and metabolic surgery in obese patients is comprehensive. Medical institutions and clinical medical and nursing staff can combine the institutional environment to transform and apply the evidence.

Objective: To understand the occupational hazard and distribution of silica dust (free SiO₂≥10%) in the workplace environment of the enterprises in Fengxian District, and to provide scientific basis for improving the working environment and protecting the physical and mental health of the workers. Methods: Individual sampling monitoring and on-site labor hygiene investigation were conducted on 421 workers involved in 87 silicon dust enterprises in the jurisdiction from 2014 to 2018, and measured concentration-time weighted average (C_TWA) . Results: The results showed that the range of the C_TWA was (0.021~17.000) mg/m³, the median was 1.600 mg/m³, and the qualified rate of 30.88%. The difference of total dust concentration was statistically significant in different years (Z=38.831, P<0.05) . The qualified rate of small-scale enterprises is higher than that of medium-scale enterprises (χ²=9.472, P<0.05) . The qualified rate of other domestic enterprises is higher than that of private enterprises and foreign enterprises (χ²=10.089, P<0.05) . The acceptance rate of metal products manufacturing is lower than that of general equipment manufacturing and other manufacturing enterprises (χ²=64.626, P<0.05) . The qualification rate of natural ventilation is higher than that of mechanical ventilation (χ²=6.278, P<0.05) . Conclusion The enterprises involved in silicon production in Fengxian District need to further strengthen the production process reform and improve the ventilation and dust removal protection measures. Widely carry out the publicity of occupational disease prevention and control law, conduct targeted pre-job training, improve workers' awareness of self-protection, and protect the occupational health of workers in many ways.

Objective To discuss the influence of preoperative family purge care for the quality of life of patients with long type of congenital Hirschsprung′s disease (HD) who had enterocolitis history in neonatal period. Preoperative family purge care, which can shorten the HD postoperative treatment, improve the quality of life. Methods A total of 40 cases of patients with long type of congenital HD who had enterocolitis history in neonatal period received 1-stage radical preoperative by family phone call. Nineteen cases from January 2010 to February 2013 were as normal group and 21 cases from March 2013 to April 2016 were as improved group. Routine family purge nursing care 3-6 months were used in both the groups, while the combined nursing care of expanding anus were used in the improved group in addition. Evaluated the effects of postoperative observation indicates: the first defecation time, length of hospital stay, time needed for expanding anus, patency rate of defecation and not patency rate in 9-12 days, need enema intervention to assist defecate rate after postoperative 1 year, the recurrence of enterocolitis at 1-3 years after operation. Results The first defecation time, length of hospital stay, time needed for expanding anus were (39.15±8.23) h, (7.89±0.82)d, (5.17±0.98) min in normal group, (23.79± 7.54) h, (7.10± 0.29) d, (3.15±0.73) min in improved group, and there were significant differences between two groups (t=6.13, 5.46, 15.54, all P<0.01). The patency rate of defecation and not patency rate in 9-12 days were 12/19, 7/19 in normal group, 100.00%(21/21), 0 in improved group, and there were significant differences between two groups (χ2=9.38, P<0.01). The intervention rate of no need for enema, occasionally enema, often enema were 2/19, 12/19, 5/19 in normal group, 76.19%(16/21), 23.81%(5/21), 0 in improved group, and there were significant differences between two groups (χ2=18.25, P<0.01). There was no significant difference in the recurrence of enterocolitis at 1, 2, 3 years after operation between two groups (χ2=2.33, P>0.05). Conclusions Patient with long type of congenital HD who had enterocolitis history in neonatal period neonatal period,received family enema and expanding anus in 3- 6 months before 1-stage radical preoperative can shorten the postoperative HD treatment, improve the quality of life.

Objective To investigate the relationship between plasma docetaxel concentration and the efficacy as well as toxic and side effects in patients with breast cancer after chemotherapy.Methods Seventy -one patients with breast cancer who accepted AC sequential T chemotherapy regimen as first line treatment were selected from April 2015 to September 2015 in Hubei Provicial Tumor Hospital.The plasma concentration of docetaxel was detected by latex immunoturdidimetry after the docetaxel continuous infusion in each cycle.The three groups were assigned according to the concentration distribution of docetaxel:group A (plasma concentration docetaxel ≤ 2.0mg·h -1 · L -1 ),group B (2.1 -2.5mg·h -1 ·L -1 )and group C (≥2.6 mg·h -1 ·L -1 ).The relationship between the drug plasma concentration,therapeutic efficacy and adverse reactions in different docetaxel plasma concentration was analyzed retrospectively by Chi -square tests.Results The average plasma concentrations of docetaxel of the three groups were (1.55 ±0.36)mg·h -1 ·L -1 ,(2.28 ±0.13)mg·h -1 ·L -1 ,(2.87 ±0.38)mg· h -1 · L -1 respectively.The adverse reactions were enhanced with the increasing of docetaxel plasma concentration (χ2 =5.169, 4.463,3.630,P =0.023,0.035,0.047).The therapeutic efficacy of group C and group B was same(95.8%),which was higher than 87.0% of group A,but there was no statistically significant difference (χ2 =1.559,P =0.24). Conclusion Breast cancer patients whose plasma concentration of docetaxel is between 2.1 ~2.5mg·h -1 ·L -1 has a better prognosis,and its adverse reactions are controlled in a certain extent.

Objective To evaluate the effects of Reyanbao combined with acupiont application on pain and comfort degree of pregnant women with surgical abortion. Methods One hundred twenty pregnant women suffering from abdominal pain after surgical abortion were randomly divided into control group and treatment group:the control group was treated with conventional nursing care and the treatment group was with Chinese medicine Reyanbao combined with acupiont application from the same treatment as in the control group, pain and comfort of patients were observed after two hours of treatment. Result The pain and comfort of the two groups were statistically significant (P < 0.05). Conclusion Chinese medicine Reyanbao combined with acupiont application can effectively relieve pain after surgical abortion and improve comfort. It is a safe, effective, convenient and practical in use of traditional Chinese medicine nursing analgesia technology.

Objective:To investigate the expression of inducible costimulatory ( ICOS) and inducible costimulatory ligand ( ICOSL) on peripheral blood mononuclear cells ( PBMCs ) and their clinical relationship with rheumatoid arthritis ( RA ) patients.Methods:Peripheral blood samples were collected from 85 RA patients and 50 HC in this study.Expression of ICOS and ICOSL on PBMC from the subjects were detected by flow cytometry and real-time polymerase chain reaction( RT-PCR).The alteration of ICOS and ICOSL were observed after hormone therapy in 15 patients with RA and the relationship between their expression level and patients′clinical manifestations were analysed.Results:The ICOS and ICOSL mRNA level of RA patients′PBMCs were significantly higher than that in HC.The expression level of ICOS on CD4+T cells was higher than than that in HC[(7.08±4.72)% vs (3.01+1.39)%,P<0.0001].The expression of ICOSL on monocytes[(5.77±3.45)%vs (3.64±1.43)%,P<0.05] and B cells [(5.78± 4.52)%vs (3.97±1.63)%,P<0.05] were significantly elevated in RA patients.In RA patients with active disease,however,ICOSL expression on monocytes and B cells were increased as compared with those in inactive RA patients [ ( 5.45 ±3.50 )% vs ( 4.04 ± 1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Furthermore,after receiving immunosuppressive therapy, the expressions of ICOS and ICOSL were notably reduced as compared with pre-therapy levels on PBMCs from patients [ ( 5.45 ±3.50)%vs (4.04±1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Conclusion:The high levels of ICOS and ICOSL expression were closely correlated with the degree of disease and therapeutic response,suggesting that ICOS/ICOSL pathway may play a critical role in pathogenesis of RA.

OBJECTIVETo investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenchymal stem cell (MSCs) on graft-versus-host disease (GVHD).

METHODSLentiviral vector containing CXCR4 was constructed. CXCR4 overexpressed MSC by lentiviral vector mediated were assessed. A major histocompatibility complex (MHC)-mismatched mouse model of bone marrow transplantation (BMT) from C57BL/6 donors to BALB/c recipients was constructed. Mice were divided into five groups: total body irradiation (TBI) group, mice received irradiation only; BMT group, mice were transplanted with bone marrow (BM) after TBI; GVHD group, mice were transplanted with BM and splencytes after TBI; CXCR4-MSC group, mice were transplanted with CXCR4-MSC, BM and splencytes after TBI; EGFP-MSC group, mice were transplanted with EGFP-MSC, BM and splencytes after TBI. The survival, body weight and clinical score of GVHD in transplanted mice were monitored. Liver, intestine and skin from mice in each group were obtained for histological examination. Plasma concentrations of inflammation factors such as interleukin (IL)-2, IFN-γ and TNF-α were also determined using a cytometric bead array cytokine kit.

RESULTSAll mice in TBI group died within 14 days, while all of BMT group survived. The mean survival times for GVHD, EGFP-MSC and CXCR4-MSC groups were (17.0 ± 2.3) d, (21.7 ± 4.8) d and (30.1 ± 9.1) d, respectively. Treatment with CXCR4 over-expressing MSCs could decrease the mortality rate. All mice in each group developed clinical signs such as hunched posture, dull fur, diarrhea and weight loss. Meanwhile, histopathological findings in target organs were confirmed the presence of GVHD. While, clinical GVHD scores and histopathological scores in CXCR4-MSC group were significantly lower than that of GVHD group. Moreover, compared with control groups, the plasma IL-2, IFN-γ and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased (P<0.05).

CONCLUSIONThe results revealed that CXCR4- transduced MSCs could effectively control the occurrence of mouse GVHD following allogeneic BM transplantation.

Animals ; Bone Marrow Transplantation ; Cytokines ; Genetic Vectors ; genetics ; Graft vs Host Disease ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; immunology ; Mice ; Receptors, CXCR4 ; genetics ; immunology ; Transplantation, Homologous

Objective To explore prevention of cyclosporine A (CsA) combined with Cobalt protoporphyrin (CoPP) against murine graft versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods C57BL/6 (H-2Kb) mice were used as donors and BALB/c (H-2Kd) mice as recipients,which were randomly divided into 4 groups.The mice in total body irradiation group (TBI group) were lethally irradiated and injected intravenously with PBS; The mice in Allo-HSCT group (BS group) were lethally irradiated and injected intravenously with bone marrow cells and spleen cells; The mice in CsA intervention group (CsA group) were injected with CsA intraperitoneally after allo-HSCT; The mice in CsA combine with CoPP intervention group (combination group) received both CsA and CoPP intraperitoneally after alloHSCT.Recipients were monitored for condition,survival rate and weight.The liver,small intestine and skin in the recipients were gained and pathological changes of GVHD were assessed.The kidney was stained with Masson staining dye to observe the tissue fibrosis.The expression levels of renal HO-1 mRNA in the recipients were detected.Results In contrast to BS and CsA groups,GVHD degree in combination group was mild,with less reduction and quick recovery of weight.On the day 30 after HSCT,survival rate in BS group was 36.8％,and that in combination group and CsA group was 69.6％ and 53.5％ respectively (P＜0.05).In comparison with BS and CsA groups,pathological changes in combination group were mild,cellular edema and degeneration degree of the liver,small intestine and skin were slight,and few necrosis and infiltrated inflammatory cells were observed.Tubulointerstitial fibrosis hardly occurred in combination group,but it occurred in CsA group abundantly.As compared with BS group,the expression levels of HO-1 mRNA was increased in combination group,while decreased in CsA group (P＜0.05).Conclusion CsA combined with CoPP enhanced the protective effect of CsA against GVHD,moreover,CoPP could alleviate the side effects of CsA,which might be related with up-regulation of the expression levels of HO-1.

Objective To identify a aldehyde dehydrogenases positive (ALDH+) cancer stem cell subpopulation in MCF-7 cells and to investigate the proliferation and differentiation characteristics of these cells in vitro.Methods ALDH+ breast cancer stem cells were isolated from MCF-7 cells by flow cytometry and the biological property of ALDH+ breast cancer stem cells were examined by scarification test,MTT,growth curvature and Transwell migration assay.Results The ratio of CD-/low24 CD+44 cells was about 1.4 ％ in MCF-7 cells.The ratio of ALDH+ CD-/low24CD+44 cells was about 1.2 ％.The growth curvature of ALDH+ breast cancer stem cells was almost the same with that of CD-/low24 CD+44 cells.The distance between cells was obviously shorter in both CD-/low24 CD+44 cells scarification zone and ALDH+ CD-/low24 CD+44 cells scarification zone.The migration ability of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells was stronger than control group cells.There were migration ability differences between CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells.The results of Transwell experiments were in coincidence with above results.Lots of CD-/low24CD+44 cells and ALDH+ CD-/low24CD+44 cells were through the membrane.In MTT assay,absorbance values were 1.05±0.098,1.56±0.075 and 1.67±0.032.Conclusion CD--/low24CD+44 and ALDH+ CD-/low24CD+44 breast cancer stem cell subpopulation exist in MCF-7 cells.ALDH could potentially be used as a molecular marker to identify breast cancer stem cell subpopulation.