OBJECTIVE:To establish the HPLC fingerprints for Herba clematidis in northeast. METHODS:HPLC was per-formed on the column of Hedera ODS-2 C18 with mobile phase of acetonitrile-0.5% phosphoric acid solution(gradient elution)at a flow rate of 1.0 mL/min,detection wavelength was 338 nm,column temperature was 30 ℃,and the injection volume was 20 μL. Using rutin as as a reference,the HPLC profiles of 10 batches of H. clematidis were determined,Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine(2004A edition) was used for the common peaks identification and similarity evaluation. RESULTS:There were 16 common peaks in the 10 batches of H. clematidis,similarity degree was higher than 0.9. It was proved that the HPLC profiles and control fingerprint profile of 10 batches of H. clematidis had good consistency. CONCLUSIONS:The established fingerprints can provide reference for the identification and quality evaluation of H. clematidis in northeast.

ObjectiveTo investigate the inhibitory effect of lentivirusly-mediated ObR-siRNA on transplanted MCF-7 human breast cancer cells by intratumoral injection.MethodsA model of subcutaneous implanted tumor was generated through injecting MCF-7 human breast cancer cells into the nude mice.Thirty established mice with MCF-7 breast cancer cells xenograft were divided into 3 groups randomly,and mice in the experimental group were intratumorally injected with ObR-siRNA lentivirus,while the negative control group and blank control group mice were injected with the same dose of negative lentivirus and normal saline.All mice were subcutaneously injected with recombinant human leptin around the tumor site once a day.Tumor size was blindly measured every other day and the mRNA expression and protein expression levels of ObR in each group were determined.ResultsKnockdown of ObR-treated xenografted nude mice with a high leptin microenvironment was successfully established.Local injection of ObR-siRNA lentivirus significantly suppressed the established tumor growth in nude mice(P ＜ 0.01,P ＜0.01 ).Real time-PCR and Western blotting showed that the mRNA and protein expression of ObR was decreased in the ObR-siRNA lentivirus group( P ＜ 0.01,P ＜ 0.01 ).ConclusionsIntratumoral injection of recomhinant ObR-siRNA lentivirus inhibits the growth of MCF-7 cells xenografts in the nude mice,suggesting that ObR might represent a therapeutic target in the genotherapies of human breast cancer.

AIM: To observe the changes of morphology, the activity of myeloperoxidase (MPO) and membrane pump activities of spleen tissue in acute renal failure (ARF) rabbits, and to inquire into the role of spleen on pathogenesis of immune function disorders during ARF. METHODS: 42 rabbits were divided into control group, HgCl_2 group and glycerinum group. The ARF model was established by hypodermic injection of 1% HgCl_2 at dose of 1.3 mL/kg in HgCl_2 group, intramuscularly injection of 50% glycerinum at dose of 10 mL/kg in glycerinum group, respectively, and the animals were divided into the 12 h, 24 h, 48 h secondary groups (6 rabbits each group). At different time points, the rabbits were cannulated to facilitate the collection of blood sample to examine the biochemical indexes of renal function. The spleen microscopic sections were prepared for observing the morphology. The spleen homogenate was made for determining the activities of MPO and membrane pumping. RESULTS: Pathological sections of spleen showed that the different degree of congestion was found and spleen trabecula was increased in two model groups at multiple-time points. The MPO activity of spleen homogenate in HgCl_2 group and glycerinum group at all time points were obviously higher than that in control group, and at 24 h, the MPO activitie in two model groups was significantly increased than that in the same group at 12 h and 48 h. The activities of Na~+-K~+-ATPase, Ca~(2+)-ATPase, Mg~(2+)-ATPase, Ca~(2+)-Mg~(2+)-ATPase of spleen homogenate in two model groups at multiple time points were significantly lower than those in control group. Following ARF development, the ATPase activitie in two model groups at 48 h was lower than that at 12 h except the Mg~(2+)-ATPase in glycerinum group. CONCLUSION: Spleen as an immune organ has histological damage, arrest of polymorphonuclear neutrophils and dysfunction of membrane pump during the development of ARF in rabbits, leading to immune disorders.

Objective To study the prevalence trends and etiology of hand,foot,and mouth disease (HFMD) in hospitalized children.Methods The clinical data of 11 510 cases of children hospitalized with HFMD from 2008 to 2017 in Department of Infection Diseases of Kunming Children's Hospital were collected,and to retrospectively analyze the characteristics,time distribution and pathogen distribution of the cases.Results Of the 11 510 children with HFMD,6 100 were male and 5 410 were female.There were 9 814 cases under 3 years old,1 696 over 3 years old.HFMD occurred throughout the year.The peak months of the disease were April to July,with the time distribution of single peak.There were 4 690 severe cases and 3 452 critical cases,accounting for 70.34％.The main pathogens detected were enteroviruses A71 (EV-A71),coxsackievirus A16 (CV-A16) and other enteroviruses (EV),with 3 803 cases (36.02％),1 122 cases (10.63％) and 3 401 cases (32.21％) respectively.EV-A71 and CV-A16 infections dominated from 2008 to 2013,while EV-A71 and other EV infection dominated from 2014 to 2017.Conclusions EV-A71,CV-A16 and other EV are the main pathogens of HFMD in Kunming.Critical HFMD cases are mainly caused by EV-A71 infection.

Objective: To explore the treatment technique, occurrence and development patterns of such radiation injuries as in a major radiological accident in which a victim suffered mild bone marrow radiation sickness combined grade degree Ⅲ acute radiation induced skin injury, based on his dose estimation, clinical manifestations and disease treatments. Methods: History inquiry in detail, earlier physical dose estimation and biological dose estimation were conducted in conjunction with analyzing the chromosome aberration of peripheral blood lymphocytes. The physical dose was estimated by Monte Carlo method.The systematic laboratory and imaging examination was performed to evaluate the condition. The comprehensive analysis was conducted to determine the diagnosis and treatment plan. Results: At 3d after the exposure, "Ren" felt mild pain and discomfortable on the skin of the right index finger. The body of the right hand index finger was covered with blister at 21 d after exposure.The estimation of biological dose was 0.43 Gy (95%CI: 0.31-0.58 Gy), and the physical dose was estimated to be 36-164 Gy for each part of the right hand finger. The hematopoietic system, immune system and endocrine system were normal. The liver function index value was transiently increased. The liver damage resulted from the use of antibiotic-induced combined with the patient′s past medical history and admission examination result, and the relevant antibiotics were discontinued. The liver function returned to normal after liver protection treatment. At 22 d after irradiation, a right finger incision and decompression surgery were performed. The stem cells were extracted and implanted into the right index finger. After 59 days of hospitalization, there was no obvious discomfort in the body, and the right index finger recovered well, as well as the pain significantly relieved, and the knuckle activity was basically normal. Conclusions The patient with excessive radiation and grade Ⅲ acute radiation skin injury was successfully treated, and local application of autologous adipose-derived stem cell transplantation achieved good result .

Objective : To explore the method of intraperitoneal injection of allogenic fecal filtrate to establish the rat model of moderate and severe sepsis． Methods: The preparation method of allogeneic fecal filtrate was determined．Allogeneic fecal filtrate of different concentrations (0. 5，1，2 g / kg) was injected intraperitoneally to observe the general situation，survival time and severe degree of sepsis of rats. After determining the optimal concentration，the success rate of the model，serum inflammatory factors，serum concentration of D-lactic acid ( D-LA) and serum intestinal fatty acid binding protein (I-FABP) ，lung function changes，lung，liver and kidney tissue injury were further observed. Results: After intraperitoneal injection of allogenic fecal filtrate for 24 h，the rats of 1 g / kg group presented fever，tachypnea and hypotension，the survival rate was 83. 3% at 24 h and 16. 7% at 48 h， 2 g / kg group rats all died within 24 h，the dose of 1 g / kg was determined for subsequent experiments．Injected fecal filtrate for 24 h，the success rate of the sepsis model was 77. 8% . The levels of interleukin-6 ( IL-6) ，tumor necrosis factor-α ( TNF-α) ，D-LA and I-FABP in serum significantly increased． There were severe edema and bleeding in lung tissue，Pulmonary function appeared respiratory dysfunction，included functional residual capacity (FRC) ，quasi static compliance ( Cdyn) ，forced expiratory volume for the first 100 milliseconds(FEV100) ，peak expiratory flow (PEF) decreased，airway resistance (RI) ，inspiratory capacity (IC) increased．Liver and kidney tissues also showed varying degrees of edema and inflammatory cell infiltration，the levels of alanine aminotransferase (ALT) ，aspartate aminotransferase (AST) ，blood urea nitrogen (BUN) and creatinine ( Cr) in serum significantly increased． Conclusion Intraperitoneal injection of allogenic fecal filtrate ( 1 g / kg) can produce a relative typical septic model in rats.