Malignant tumors in farmers over 60 years old of the high-risk population in a town in Shanghai / 上海预防医学

Objective To screen for malignant tumors and high-risk factors in rural residents over 60 years old， so as to prevent and control the occurrence and development of tumors in the future. Methods The survey was conducted with reference to part of the questionnaire in the "Urban Cancer Early Diagnosis and Treatment Project and Evaluation of High-risk Populations". Clinical examinations included serum tumor marker detection， CT screening for lung cancer， occult blood （+） plus colonoscopy screening for colorectal cancer， and mammography screening. Individuals who were positive in the abovementioned clinical tests were defined as high-risk subjects. Results A total of 271 high-risk subjects （1.91%） were screened out of 14 161. Among the high-risk subjects， 71 cases of malignant tumors （26.19%） were found， with an incidence rate of 501.38 per 105. The top five tumors （63.38% of all diagnosed） were mainly concentrated in lung， upper digestive tract， blood system， urinary system， and rectum-colon. The proportion of malignant tumors detected by positive indicators was 61.54% of blood； 46.15% of carcinoembryonic antigen and carbohydrate antigen 125； 23.08% of alpha-fetoprotein； 16.66% of lung CT； and 3.09% of prostate PSA. The positive indicators in the high-risk subjects were mainly for the tumors in the prostate， lungs， liver， and CEA/CA125. The subjects with positive test indicators had lower average annual income in the last 5 years than the normal subject group （χ2=3.380， P=0.040）. The subjects with positive test indicators had higher proportion in family history of tumors than the normal group （χ2=2.596， P=0.046）. People in thehigh-risk group had a higher proportion than the normal group in suffering from hypertension， liver disease， gastrointestinal disease， respiratory system disease， and surgical treatment. Patients with high-risk tumors were found to have higher proportion than the normal group in showing pre-tumor clinical symptoms in the last 1 year. Study of the tumor-related risk factors found that the high-risk group had a higher proportion of high-fat/high-cholesterol diet， alcohol drinking， passive smoking， and personality depression. Conclusion High tumor risk factors have been identified in this population. It is necessary to strengthen the corresponding intervention and follow-up treatment of precancerous diseases in the future. We recommend the government to conduct tumor screening among high-risk groups to improve cost-effectiveness.

Study on the treatment of thin endometrium with growth hormone and vitamin E / 中国临床药理学与治疗学

AIM: To study the effect of growth hormone (GH) and vitamin E (Vit.E) combined in the treatment of endometrial thinning. METHODS: Twenty female SD rats were randomly divided into four groups: control group, model group, GH group and treatment group, with 5 rats in each group. Control group was routinely fed; Rats in model group, GH group and treatment group were injected intrauterine with 95%ethanol during estrus stage to construct a thin endometrial model. Six to eight hours after operation, rats in model group were injected intrauterine with 0.2 mL normal saline, rats in GH group and treatment group were injected with the same amount of GH, and the treatment group was given intragastric treatment of 60 mg/kg Vit.E. The rats were sacrificed 3 estrus cycles (about 2 weeks) after the operation. HE staining was performed on the uterine tissue to identify the model, and the levels of Cytokeratin 19 and Vimentin in the endometrium were detected by immunohistochemical color. RESULTS: The endometrial thickness of the model group was significantly thinner than that of the model group, and the endometrial thickness of the treatment group was significantly higher than that of the control group, but the endometrial thickness of the GH group was slightly lower than that of the control group. The expression levels of keratin and vimentin in model group were lower than those in GH group, control group and treatment group, and the differences were statistically significant. CONCLUSION: Endometrial-related proliferation indexes were significantly increased after GH and vitamin E treatment, and GH and vitamin E could effectively promote the proliferation of endometrial cells.

Reduced inflammatory responses to SARS-CoV-2 infection in children presenting to hospital with COVID19 in China

BackgroundInfection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in children is associated with better outcomes than in adults. The inflammatory response to COVID-19 infection in children remains poorly characterised. MethodsWe retrospectively analysed the medical records of 127 laboratory-confirmed COVID-19 patients aged 1 month to 16 years from Wuhan and Jingzhou of Hubei Province. Patients presented between January 25th and March 24th 2020. Information on clinical features, laboratory results, plasma cytokines/chemokines and lymphocyte subsets were analysed. FindingsChildren admitted to hospital with COVID-19 were more likely to be male (67.7%) and the median age was 7.3 [IQR 4.9] years. All but one patient with severe disease was aged under 2 and the majority (5/7) had significant co-morbidities. Despite 53% having viral pneumonia on CT scanning only 2 patients had low lymphocyte counts and no differences were observed in the levels of plasma proinflammatory cytokines, including interleukin (IL)-2, IL-4, IL-6, tumour necrosis factor (TNF)-, and interferon (IFN)-{gamma} between patients with mild, moderate or severe disease. InterpretationsWe demonstrated that the immune responses of children to COVID-19 infection is significantly different from that seen in adults. Our evidence suggests that SARS-CoV-2 does not trigger a robust inflammatory response or cytokine storm in children with COVID-19, and this may underlie the generally better outcomes seen in children with this disease. These data also imply anti-cytokine therapies may not be effective in children with moderate COVID-19. FundingThis study was funded by National Natural Foundation of China (No. 81970653). Research in contextO_ST_ABSEvidence before this studyC_ST_ABSWe searched PubMed without language restriction for studies published until June 25, 2020, using the search terms "SARS-CoV-2" or "novel coronavirus" or "COVID-19" and "immune responses" or "innate immunity" or "cytokine" or "subset of lymphocyte" and "children" or "adolescent". Previously published research describes that severe and fatal cases in children are relatively rare. However, the inflammatory responses to COVID-19 infection in children remains poorly characterised. Added value of this studyWe analysed data from 127 laboratory-confirmed COVID-19 patients aged 1 month to 16 years in Hubei province to explore the immune responses to SARS-CoV-2 infection presenting to hospital with COVID-19. Cell numbers of CD3+, CD4+, CD8+ and natural killer T cells were within mostly normal limits even in more severe cases, and the levels of immunoglobulins, and proinflammatory cytokines, including interleukin (IL)-2, IL-4, IL-6, tumour necrosis factor (TNF)-, and interferon (IFN)-{gamma} were not generally elevated regardless of disease severity. Implications of all the available evidenceThe immune response to SARS-CoV-2 infection of children is significantly different from that seen in adults. The inflammatory responses seen even in children with viral pneumonia on CT are relatively mild and do not trigger the "cytokine storm" seen in some adults with COVID-19. This implies anti-cytokine therapies may not be effective in children with COVID-19.

The Observation of Clinical Effect of Using Ambroxol in the Treatment of Acute Exacerbation of Chronic Obstructive Pulmonary Disease / 昆明医科大学学报

Objective To compare the efficacy of two doses of ambroxol in the treatment of the elderly patients with acute exacerbation of chronic obstructive pulmonary disease (COPD).Methods A hundred and ten cases of the elderly patients with an acute exacerbation of COPD were divided into low-dose group and high-dose group.Low-dose group (56 cases) were given Ambroxol 30 mg,2 times per day for 10 day and high-dose group (54 cases) were given Ambroxol 60 mg,2 times per day for 10 days.Results The effective rate in high-dose group was significantly higher than that in low-dose group.Conclusion High-dose ambroxol has significant effect in the treatment of acute exacerbation of chronic obstructive pulmonary disease.

Study on anti ovarian cancer efficacy elicited by CD34~+ umbilical cord blood hematopoietic stem cells transfected with IL-21 in nude mouse model / 中华微生物学和免疫学杂志

Objective To investigate the CD34~+ umbilical cord blood hematopoietic stem cells (CD34~+ UBSC) transfected with interleukin 21 (IL-21) against the ovarian cancer effect in tumor-bearing nude mice. Methods CD34~+ UBSC were obtained from the UBSC by a magnetically activated cell sorting technique and then transfected with recombinant plasmid plRES2-IL-21-EGFP after the CD34~+ UBSC were proliferated in vitro. The therapeutic effect was evaluated by the size of the tumor and the life span in nude mice treated with the CD34~+ UBSC-IL-21. The expression of IL-21 and its bioactivity in CD34~+ UBSC-IL-21 and in local neoplasitc tissues were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR), immune fluorescence technique, ELISA, Western blot, immunohistochemistry and splenocyte proliferative activity. The NK cell cytotoxicity and the numbers of NK cells, serum level of IFN-γ and TNF-αwere simultaneouly detected by FCM and ELISA, respectively. Results CD34~+ UBSC were cultured and transfected with pIRES2-IL-21-EGFP successfully. CD34~+ UBSC-IL-21 could inhibit the tumor growth and extended nude mice life span compared with other groups (P < 0.01). The expression of IL-21 in the neo-plastic tissue, serum level of IFN-γ and TNF-α , NK cell activity and the numbers of NK cells of mice origin and of human origin in splenocytes were increased significantly in the nude mice treated with CD34~+ UBSC-IL-21 compared with other groups(P <0.01). Conclusion The CD34~+ UBSC transfected with IL-21 have competent against ovarian cancer in tumor-bearing nude mice. The findings may establish a foundation for gene therapy of the ovarian cancer by CD34~+ UBSC-IL-21 in clinic application.

Construction of Sox9 gene eukaryotic expression vector and its inductive effects on directed differentiation of bone marrow stromal cells into precartilaginous stem cells in rats / 华中科技大学学报（医学）（英德文版）

Sox9 gene was cloned from immortalized precartilaginous stem cells and its eukaryotic expression vector constructed in order to explore the possibility of bone marrow-derived stromal cells differentiation into precartilaginous stem cells induced by Sox9. A full-length fragment of Sox9 was obtained by RT-PCR, inserted into pGEM-T Easy clone vector, and ligated with pEGFP-IRES2 expression vector by double digestion after sequencing. The compound plasmid was transfected into born marrow-derived stromal cells by Lipofectamine 2000, and the transfection efficacy and the expression of Sox9 and FGFR-3 were observed. Flow cytometry was used to identify the cell phenotype, and MTT was employed to assay proliferative viability of cells. Sequencing, restrictive endonuclease identification and RT-PCR confirmed that the expansion of Sox9 and construction of Sox9 expression vector were successful. After transfection of the recombinant vector into bone marrow-derived stromal cells, the expression of Sox9 and FGFR-3 was detected, and proliferative viability was not different from that of precartilaginous stem cells. It was concluded that Sox9 gene eukaryotic expression vector was successfully constructed, and the transfected bone marrow-derived stromal cells differentiated into the precartilaginous stem cells.

Separation,identification and immortalization of precartilaginous stem cells from neonatal rats / 中国组织工程研究

BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.

Anti-tumor mechanisms of lymphopenic mice transferred with tumor-specific lymphocytes and immunized with mIL-21 tumor vaccine / 中华微生物学和免疫学杂志

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.

Effect of Huangqi Injectio on Abnormal Expression of Aquaporin-1 Caused by Glucose Lactate Peritoneal Dialysis Solution / 中医杂志

Objective To observe the effect of Huangqi (Radix Astragali seu Hedysari) Injectio on the abnormal expression of aquaporin-1 (AQP-1) caused by glucose lactate peritoneal dialysis solution. Methods Different glucose concentration lactate peritoneal dialysis solutions and Huangqi Injectio given to the rats intraperitoneally. Eight weeks later, the ultrafiltration of peritoneal dialysis, peritoneum clearance, peritoneal structure, AQP-1 and its gene expression were observed. Results Compared with the control group, in the glucose and Huangqi groups, the ultrafiltration of peritoneal dialysis was remarkably decreased (P