BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

AIM To observe the protective effects of Qigu Capsule-medicated serum on C2C12 myotube cell atrophy induced by dexamethasone.METHODS The Qigu Capsule-medicated serum was prepared.C2C12 cells cultured in vitro were divided into the normal control group,the dexamethasone group,the 10%blank serum group and the 10%Qigu Capsule-medicated serum group for 48 h,corresponding drug treatment,followed by 24 h culture with 10 μmol/L dexamethasone except the control group,and had cell viability detection by CCK-8 method.With their myotube cells intervened accordingly by the aforementioned regime following 6-7 days differentiation with 2%horse serum induction,the C2C12 cells had their myotube cells diameter measured;and the expressions of proteins related to MyHC,MuRF-1,Atrogin-1 and PI3K/Akt signaling pathway detected by Western blot.The impact of PI3K inhibitor LY294002 on the diameter of myotube cells and the expression of MuRF-1,Atorgin-1 and PI3K/Akt signaling pathway related proteins were detected as well.RESULTS Compared with the normal control group,the dexamethasone group and the blank serum group were observed with decreased viability of C2C12 cells(P<0.01),decreased diameter of myotube cells(P<0.01),increased protein expressions of MuRF-1 and Atrogin-1(P<0.01),and decreased expression of MyHC protein and phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.01).Compared with the dexamethasone group,the Qigu Capsule-medicated serum group showed increased viability of C2C12 cells(P<0.01);increased diameter of myotube cells(P<0.01);decreased protein expressions of MuRF-1 and Atrogin-1(P<0.01);and increased expression of MyHC protein and phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.05,P<0.01).The intervention of LY294002 upon the Qigu Capsule-medicated serum group offset the anti-muscular tube atrophy effect(P<0.01),increased the protein expressions of MuRF-1 and Atorgin-1(P<0.01),and decreased the phosphorylation levels of PI3K,Akt,mTOR and FoxO3a(P<0.05,P<0.01).CONCLUSION The Qigu Capsule-medicated serum can alleviate the atrophy of C2C12 myotubes induced by dexamethasone,and its mechanism may be related to the activation of PI3K/Akt signaling pathway.

Child ; Humans ; Rural Population ; China/epidemiology* ; Respiratory Tract Infections/prevention & control*

Humans ; Neurons ; Olivary Nucleus ; Superior Olivary Complex

Objective:To explore effects of telephone transitional nursing after discharge from hospital on treatment effectiveness and quality of life of patients with lymphedema after breast cancer surgery.Methods:According to the convenient sampling method, a total of 85 patients with lymphedema after breast cancer surgery who were admitted to Department of Gynecology in Wuxi Maternal and Child Health Care Hospital from October 2017 to October 2019 were selected as research objects. According to the double-blind principle, the selected objects were divided into the study group ( n=43) and the control group ( n=42) . After discharge from hospital, patients in the study group were given telephone follow-up while patients in the control group were given routine follow-up. The effects of different methods on the quality of life and treatment effectiveness of patients with lymphedema after breast cancer surgery were compared. Results:After the follow-up, the difference in circumference of upper limbs of the study group was lower than that of the control group, and the difference was statistically significant ( P<0.01) . The anterior flexion and abduction of the shoulder joint of the study group were higher than those of the control group, and the differences were statistically significant ( P<0.01) . The total effective rate of treatment in the study group was higher than that in the control group, and the difference was statistically significant ( P<0.05) . After follow-up, except for role-physical, the quality of life scores in the study group were higher than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The application of telephone transitional nursing after discharge from hospital in patients with lymphedema after breast cancer surgery can extend the nursing service, promote the recovery of shoulder joint function, ensure the effectiveness of treatment and improve the quality of life of patients, which is suitable for clinical promotion and application.

OBJECTIVE: To follow up the participants of the randomized clinical trial "Efficacy and Safety of Niaoduqing Particles () for Delaying Moderate-to-Severe Renal Dysfunction", and assess the long-term effects of Niaoduqing Particles on delaying the progression of renal dysfunction. METHODS: Participants, who had previously been randomly assigned to receive Niaoduqing Particles or placebo for 24 weeks (146 cases in each group), were invited to follow-up and all were administered Niaoduqing Particles 5 g thrice daily and 10 g before bedtime for 24 weeks. The primary endpoints were changes in baseline serum creatinine (Scr) and estimated glomerular filtration rate (eGFR) after completion of the open-label treatment period. RESULTS: After the double-blind period, the median (interquartile range) changes in Scr were 1.1 (-13.0-24.1) and 11.7 (-2.6-42.9) μmol/L for the Niaoduqing Particle and placebo groups, respectively (P=0.008), and the median changes in eGFRs were-0.2 (-4.3-2.7) and-2.21 (-5.7-0.8) mL•min•1.73 m, respectively (P=0.016). There were significant differences in the double-blind period changes in renal function between groups. After the open-label period, the median changes in Scr were 9.0 (-10.0-41.9) and 17.5 (-6.0-50.0) μmol/L for the Niaoduqing Particle and placebo groups according to baseline grouping, respectively (P=0.214), and the median changes in eGFRs were-2.3 (-6.4-1.9) and-3.7 (-7.5-1.1) mL•min•1.73 m, respectively (P=0.134). There were no statistical differences in the open-label period changes in renal function between groups. The eGFR reduction of participants who accepted Niaoduqing Particle treatment for 48 weeks was projected to 2.5 mL•min•1.73 m per year. CONCLUSION Niaoduqing Particles appear to have long-term efficacy for patients with moderate-to-severe renal dysfunction. Although there was no statistical difference, the early use of Niaoduqing Paticles seems to ameliorate the worsening of renal function. (Trial registration No. ChiCTR-TRC-12002448).
Adult ; Disease Progression ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Glomerular Filtration Rate ; drug effects ; Humans ; Kidney Diseases ; drug therapy ; physiopathology ; Male ; Middle Aged ; Outcome Assessment (Health Care)

Objective To understand the infection status and risk factors of Entamoeba histolytica in inpatients in Tengchong City, Yunnan Province. Methods A cross-sectional study was conducted in the inpatients in Tengchong People’s Hospital, Yunnan Province. After obtaining the informed consent from the subjects, the stool samples were collected from 2016-07-01 to 2017-03-31, and nested polymerase chain reaction (PCR) was used to detect E. histolytica in the stool samples. Meanwhile, a structured questionnaire was used to record the demographic information and clinical symptoms for the patients. Results Totally 507 cases were recruited, and the detection rate of E. histolytica was 1.97% (10/507, 95% CI: 1.07%-3.59%) in all subjects. There were no significant differences between the inpatients with and without E. histolytica infection in the height (Z = -0.40, P = 0.69), weight (Z = -0.34, P = 0.73), body mass index (Z = -0.40, P = 0.69) and age (Z = -1.48, P = 0.14). Chronic diarrhea (OR = 21.43, 95% CI: 5.04-91.23) and daily drinking water (OR = 11.28, 95% CI: 2.79-45.56) were relevant to E. histolytica infection. No significant association was observed between E. histolytica infection and the clinical symptoms, such as abdominal distension (OR = 0.70, 95% CI: 0.09-5.56), inappetence (OR = 0.50, 95% CI: 0.06-4.02), itchy skin (OR = 0.79, 95% CI: 0.10-6.38), perianal pruritus (OR = 1.74, 95% CI: 0.21-14.07), and constipation (OR = 0.91, 95% CI: 0.13-7.33). Conclusion E. histolytica infection is high in inpatients in Tengchong City, Yunnan Province, and chronic diarrhea and drinking unboiled water were highly correlated with E. histolytica infection.

Objective To investigate antibiotics resistance of Proteus mirabilis isolated from stools of patients with acute diarrhea for the prevention and treatment of its infection and the rational use of antibiotics. Methods Stool samples of acute diarrhea patients were collected in the diarrhea outpatient clinic of the Second Hospital of Tianjin Medical University and Tianjin Medical University General Hospital from 2013 to 2014. Enrichment culture and biochemical identification were used to isolate and identify Proteus mirabilis, which were further performed antimicrobial susceptibility testing and class 1 integron detection. Extended spectrum β-lactamases (ESBLs) phenotype and ESBLs genes (TEM, OXA and CTX-M) were amplified by polymerase chain reaction (PCR), and sequencing were carried on in parts of suspected isolates. ESBLs-positive strains were analyzed by pulsed-field gel electrophoresis (PFGE). Results A total of 277 strains of non-repetitive Proteus mirabilis were isolated, and 268 of them were performed antimicrobial susceptibility testing (the remaining 9 strains failed to recover). Relative higher resistant rates were trimethoprim/sulfamethoxazole (30.2%), ampicillin (25.4%), nalidixic acid (25.7%), streptomycin (21.6%) and chloramphenicol (21.3%). The multiple drug resistance rate was 24.6% (66/268). The positive rate of class 1 integron was 22.8%(61/268). Resistance rates to third-generation cephalosporin, ciprofloxacin and imipenem were less than 10%, but 4 isolates were resistant to imipenem, third-generation cephalosporin, fluoroquinolones, trimethoprim/sulfamethoxazole, and chloramphenicol simultaneously. Three cefotaxime-resistant strains (1062, 1505 and 1650) were positive for ESBLs phenotype and harbored CTX-M extended-spectrum β-lactamase genes, among them 2 strains also carried TEM and/or OXA β-lactamase genes. The clustering analysis of pulsed-field gel electrophoresis (PFGE) displayed that the similarities between 1505 and 1650 were 85.7%, and the similarity with 1062 was 58.1%. Conclusion Proteus mirabilis isolated from patients with acute diarrhea in our city show significant multidrug resistance, high positive rate of class 1 integron, and emergence of ESBLs-positive strains resistant to imipenem and fluoroquinolones, which pose a threat to public health. Rational use of antibiotics is important in both clinical and nonclinical settings.