Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity. The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD). Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development, differentiation, survival and function. Meanwhile, GDNF mutations are a major cause of HD. In this study, BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3, and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM). Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5, VIP and nNOS. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs. The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM. This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders, such as HD.

OBJECTIVE:To establish a method for the content determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules. METHODS:HPLC method was adopted. The determination was performed on WondasilTM-C18 column with mobile phase consisted of acetonitrile-0.4% phosphoric acid solution(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 30 ℃,and the detection wavelength was set at 254 nm. The sample size was 5 μL. RESULTS:The linear range of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid were 0.014-0.560 μg (r＝0.999 2),0.022-0.880 μg(r＝0.999 6),0.010-0.400 μg(r＝0.999 9),0.016-0.640 μg(r＝0.999 9),0.011-0.440 μg(r＝0.999 1), respectively. The limits of quantitation were 0.752,0.647,0.483,0.372,0.242 mg/L;the limits of detection were 0.253,0.185, 0.157,0.162,0.098 mg/L. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The average recoveries were 97.4%-100.9%(RSD＝1.4%,n＝6),95.7%-99.0%(RSD＝1.1%,n＝6),95.1%-98.1%(RSD＝1.3%,n＝6),96.4%-99.8%(RSD＝1.4%,n＝6),97.8%-100.4%(RSD＝1.0%,n＝6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity.The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD).Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development,differentiation,survival and function.Meanwhile,GDNF mutations are a major cause of HD.In this study,BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3,and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM).Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5,VIP and nNOS.Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs.The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM.This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders,such as HD.

Objective:To explore the surgical procedure and clinical effect of the free Flow-through superficial peroneal artery flap for repairing the digit injury with defect of proper palmar digital artery.Methods:From June, 2015 to December, 2019, free Flow-through superficial peroneal artery flap was used to repair the digit injury and to bridge the proper digital artery in 7 digits of 7 patients. There were 2 thumbs, 3 index fingers and 2 middle fingers. The size of defects on digits ranged from 2.5 cm×3.0 cm to 5.0 cm×7.0 cm. The defects of digit proper artery were 1.0 to 3.0 cm. The size of flaps was from 3.0 cm×3.5 cm to 5.5 cm×8.0 cm. The donor areas of the flap were directly sutured or covered with skin graft. Postoperative supportive treatments were given, such as infection prevention, antispasmodic, anticoagulant, analgesia and fluid infusion. The patients were kept in bed for 1 week after surgery. Monthly follow-up review were conducted and the patients were kept with the rehabilitation exercises under medical guidance.Results:All the patients entered postoperative followed-up for 6 to 18 months, with an average of 8 months. All flaps survived without any adverse event. All wounds achieved stage-one-healing. The flaps appeared in good colour, texture, elasticity and the plumps of the digit without obvious bloating. There was no obvious swelling and atrophy of the digits. The skin temperature was normal. According to the Standard for Evaluation of Upper Limb Function by the Hand Surgery Society of Chinese Medical Association, 3 digits were excellent and 4 were good. There was no obvious scar at the donor site of shank. The donor site had a good appearance and the limb function was not affected.Conclusion:The free Flow-through superficial peroneal artery flap is an ideal material to repair the defect of digit with the defect of proper digital artery. It has the advantages of simple surgical procedure, reliable blood supply and satisfactory appearance. The defect of proper digit artery can be repaired at the same time of the surgical procedure. The blood supply, appearance and function of the digits could be well recovered and the donor site is not affected.

Objectives To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza.Methods This was a randomized,double-blind,double-dummy,placebo and positive control,multicenter clinical trial,comparing peramivir trihydrate with oseltamivir and placebo.The inclusive criteria were 15-70 years old,onset within 48 h,positive rapid influenza antigen test,and febrile(＞38℃) accompanied with at least two associated symptoms.The severe cases complicated with chronic pulmonary and cardiac diseases,malignancies,organ transplantation,hemodialysis,uncontrolled diabetes,immunocompromised status,pregnancy and coexistence of bacterium infections were excluded.All patients were randomized 2:2:1 to receive peramivir,oseltamivir and placebo respectively.The primary endpoint was the disease duration,the secondary endpoints included time to normal axillary temperature and normal living activities,viral response,and adverse effects.Results Following informed consent,133 patients were included in this study.Four patients were exclude due to missing medical records,not fitting inclusion or exclusion criteria and poor compliance.A total of 129 patients were finally analyzed,including 49 cases,54 cases and 26 cases in peramivir group,oseltamivir group and placebo group.The median disease duration were 96 (76,120)hours,105(90,124) hours,and 124 (104,172)hours in three groups respectively(P＞0.05).The time to normal axillary temperature,normal living activities and viral response were not significantly different in three groups(P＞0.05).Conclusion The value of antiviral therapy in patients with mild influenza needs to be further determined.

OBJECTIVE To provide a reference for the definition of the legal attributes of the provincial Chinese herbal pieces processing norms and the improvement of legal expression for the drug standards in Drug Administration Law. METHODS Based on the legal evolution of local drug standards in China， the composition of national drug standards for Chinese herbal pieces and the composition of current provincial Chinese herbal pieces processing norms were analyzed； at the same time， based on the method of legal interpretation， the legal connotation of the provincial Chinese herbal medicine processing norms in Drug Administration Law was investigated. RESULTS & CONCLUSIONS In the practice of drug supervision， the provincial Chinese herbal pieces processing norms are essentially local drug standards. The newly revised Drug Administration Law defines the drug standards specifically， but does not stipulate the legal attribute of drug standards for the provincial Chinese herbal pieces processing norms， and there are certain legislative technical defects. It is suggested that when amending the Drug Administration Law， the legal attributes of the drug standards of the provincial Chinese herbal medicine processing norms should be clarified.

OBJECTIVE To sort out and analyze the expression and provisions of drug standards in the text of the newly revised Drug Administration Law ，and to explore the connotation and legal positioning of drug registration standards so as to provide reference for the rational application and interpretation of relevant provisions of drug standards in Drug Administration Law . METHODS Through the review of the evolution of drug standard management in China ，the legal provisions of drug standard in the Drug Administration Law were analyzed. Comparative study and literature research methods were used to analyze the legal expression and connotation of drug standards. RESULTS & CONCLUSIONS There were different expressions about “national drug standards ”“drug standards ”and“quality standards ”in the current provisions of the newly revised Drug Administration Law ； the legal position of the provincial-level standard for the preparation of Chinese herbal pieces was not clear ，and there may be insufficient legal regulation in the enforcement of drug administration. It is necessary to make an administrative interpretation for the content of relevant drug standards and provisions ，and further clarify the legal attributes of drug standards in the processing of provincial Chinese herbal pieces in order to promote the standardized management of Chinese herbal pieces.