BACKGROUND:Chronic hepatitis B virus infection can impact the biological characteristics of bone marrow mesenchymal stem cel s. Adipose-derived mesenchymal stem cel s have gained more and more attention due to their high safety, little invasiveness, easy purification and rapid proliferation. OBJECTIVE:To establish the isolation and culture method of adipose-derived mesenchymal stem cel s from patients with hepatitis B virus infection in vitro, and to observe the biological characteristics of cel s. METHODS:Adipose-derived mesenchymal stem cel s were isolated from the subcutaneous fat of hepatitis B virus infection patients by col agenase digestion and adherent method. Growth curve of adipose-derived mesenchymal stem cel s were detected by MTT method and cel phenotypes were detected by flow cytometry. The adipogenic and osteogenic differentiation potential of adipose-derived mesenchymal stem cel s were detected in vitro. RESULTS AND CONCLUSION:(1) Adipose-derived mesenchymal stem cel s from 10 patients with hepatitis B virus infection were al isolated and cultured successful y. The primary passage time of adipose-derived mesenchymal stem cel s was (8.3±1.2) days. The growth curve of cel s was“S”shaped. Cel s came into a logarithmic phase at days 3-4, and came into platform at day 7. (3) Passage 3 adipose-derived mesenchymal stem cel s highly expressed CD29, CD166, HLA-ABC and CD44, but did not express or lowly expressed CD34 and HLA-DR. (3) Adipose-derived mesenchymal stem cel s differentiated into adipocytes after adipogenic induction, and differentiated cel s were positive for oil red O staining;after osteogenic induction, adipose-derived mesenchymal stem cel s differentiated into osteoblasts that were positive for alkaline phosphatase staining. These findings indicate that the col agenase digestion and adherent method can be used to effectively isolate adipose-derived mesenchymal stem cel s from patients with hepatitis B virus infection, and the cel proliferation is rapid so that a large number of cel s can be obtained in the short term.

Objective To study the clinical effect of phentolamine combined with furosemide therapy in children with pneumonia complicated with heart failure.Methods The subjects were 92 cases of children with pneumonia complicated with heart failure in our hospital from October 2015 to December 2016,randomly divided into control group and observation group,46 cases in each group.The control group was treated with on the basis of routine treatment,the observation group,patients received phentolamine combined with furosemide treatment.Compared two groups of the total efficiency and the symptoms disappear time,and compare the cardiac function and electrocardiogram improvement effect.Results The total efficiency of observation group(93.48％)was significantly higher than the control group(71.74％)(χ2=7.57,P<0.05); observation group of the pulmonary rales disappeared time,breath,heart failure control time and hospitalization time were significantly lower than the control group(P<0.05); after treatment,the observation group of the ESV was significantly lower than the control group(P<0.05),EDV was significantly higher than that of control group(P<0.05),electrocardiogram indicators improved significantly better than the control group(P<0.01).Conclusion The clinical curative effect of phentolamine combined with furosemide in treatment of children with pneumonia complicated with heart failure significantly,can effectively improve the cardiac structureAnd function,reduce the occurrence of abnormal ECG,high safety,worthy of clinical promotion

Objective To study the effect of low molecular weight heparin calcium combined with Bailing capsule in treatment of children with Henoch-Schonlein purpura nephritis and the efficacy of serum hs-CRP, MMP-9, APN and CysC.MethodsFrom January 2013 to December 2015, 84 children with Henoch-Schonlein purpura nephritis were enrolled in our hospital.According to the order of admission, the observation group and control group were divided into observation group and control group.The control group were given low molecular weight heparin calcium, the observation group were treated with low molecular weight heparin calcium combined with Bailing capsule for treatment.The levels of serum hs-CRP, MMP-9, APN, CysC, inflammatory factors and immune levels were observed in the two groups.ResultsAfter treatment, the clinical effective rate of the observation group was significantly higher than that of the control group[92.86% (39/42) vs 69.05% (29/42)] (P<0.05).There were no significant differences in hs-CRP, MMP-9, APN and CysC levels between the two groups.After treatment, the levels of hs-CRP, MMP-9 and CysC in the two groups were significantly higher than those before treatment (P<0.05).APN was significantly higher than that before treatment (P<0.05).Compared with the control group, the levels of hs-CRP, MMP-9 and CysC in the observation group were lower than those in the control group (P<0.05), and the APN level was lower than that in the control group (P<0.05).There were no significant differences in TNF-α, IL-6, IL-10 and IL-18 levels between the two groups before treatment.After treatment, The levels of TNF-α, IL-6 and IL-18 in the two groups were significantly lower than those before treatment (P<0.05).There were no significant differences in IgA, IgG and IgM levels between the two groups.After treatment, the levels of IgA, IgG and IgM in the two groups were significantly lower than those before treatment (P<0.05).The levels of IgA, IgG and IgM in the observation group were lower than those in the control group (P<0.05).There was no significant difference in adverse reaction rate between the two groups.ConclusionLow molecular weight heparin combined with Bailing capsule can effectively reduce hs-CRP, MMP-9, CysC level and APN level in children with Henoch-Schonlein purpura nephritis, improve clinical symptoms, good clinical efficacy and high safety.

Objective To study the protective effect of emodin on hepatic ischemia-reperfusion injury in the rat(liver). Methods Thirty Wistar rats were randomly devided into 3 groups:the sham-operatment group (Control group),the ischemia-reperfusion group (I/R group) and the emodin-treatment group (EMD group). By(clamping) the left and median lobes of the liver for 1 h and followed by 1 h of reperfusion,the(partial) (70%)(normothemic)(ischemia)-reperfusion injury model was established. The rats of emodin group received emodin [60mg/(kg?d)] by(oral) route for 3 days before ischemia. Blood and liver samples were obtained for(biochemical) and(histological) analysis 1 h after reperfusion. Some liver specimens were taken for flow cytometry(analysis) to assess the(mitochondrial)(membrane) potential (??m) . Rhodanmin123 and PI were used to label the cells.Results After reperfusion,(serum) levels of alanine transaminase(ALT) and liver contents of(malondialdehyde) (MDA) in the I/R group rats were increased significantly (P

AIM: To study the changes of neurokinin A (NKA) and NKA mRNA in lung tissues of asthmatic guinea pigs exposed to chronic cigarette smoke and investigate the mechanism. METHODS: The model of asthma in guinea pigs was made by exposure to aerosolized ovalbumin and animals were randomly assigned into: ① inhale cigarette smoke (ICS) 2 weeks before provoked; ② ICS 2 weeks after provoked; ③ 2 weeks after provoked (no-ICS); ④ asthma (no-ICS before provoked); ⑤ normal control. The contents of NKA in lung tissues were detected by ELISA and NKA mRNA expression was detected by reverse transcription-polymerase chain reaction. RESULTS: ① The levels of NKA and NKA mRNA expression in lung tissues of asthma group were significantly higher than those in normal control group (P

Objective To quantitatively analyze the palpebral fissure changes in the neostigrnine tests with videonystagmography.Methods Thirty-five patients with myasthenia gravis ( MG),21 non-MG patients and 23 healthy volunteers were intramuscular injected with neostigmine methylsulfate(0.02 mg/kg) and atropine(0.5 mg).The width of palpebral fissure was measured before and after the injection for one hour( once every 10 minutes).The differences of palpebral fissure width of each time point within each group and between groups were analyzed quantitatively with repetitive measure ANOVA.Results There was a significant difference of palpebral fissures (mm) width at different time points within the MG group( before the test:4.67 ± 1.87,after the test:10 min:0.88 ±0.96,20 min:1.49 ± 1.38,30 min:1.71 ± 1.53,40 min:1.77 ± 1.82,50 min:1.79 ± 1.52,60 min:1.62 ± 1.68 ; F =11.202,P =0.002).There were also significant changes of palpebral fissures width in the MG group compared to the non-MG group and the normal control group (F =15.569,P ＜ 0.01; F =15.104,P ＜ 0.01 ).There was alsostatistical significance in the rate of change between these groups.Receiver operating characteristic analytical procedure indicated that 1.17 mm or 17.5％ was of better diagnostic capability in the neostigmine test.Conclusions Measuring palpebral fissure width with videonystagmography could be seen as an objective and accurate method.A patient with palpebral fissure width higher than 1.17 mm or 17.5％ should be highly suspected as MG.

Objective To investigate the relationship between the proportion of Th17 cells and IL-17 expression in the peripheral blood and their clinical significance in patients with non-small cell lung cancer (NSCLC).Methods The proportion of Th17 cells determined by flow cytometry (FCM) and IL-17 detected by enzyme-linked immunosorbent assay (ELISA).The data was analyzed from 60 untreated patients with NSCLC (NSCLC group) and 20 healthy volunteers (control group).Results The Th17 cells proportion in CD+3 T-cells in NSCLC group was significantly higher than that in control group [(1.23±0.41) ％ vs (1.05±0.28) ％,t =1.679,P =0.097].The expression levels of Th17 cells in peripheral blood of patients with squamous cell carcinoma was significantly higher than that in adenocarcinoma patients [(1.31±0.39) ％ vs (1.09±0.41) ％,t =2.093,P =0.041].The concentration of IL-17 of patients with NSCLC was significantly higher than that in control group [(21.11±7.87) pg/ml vs (17.64±5.07) pg/ml,t =2.280,P =0.027].The expression levels of serum IL-17 of squamous cell carcinoma patients was significantly higher than that of adenocarcinoma patients [(23.11±8.73) pg/ml vs (18.54±6.38) pg/ml,t =2.203,P =0.032].As clinical stage progressed,the proportion of Th17 cells and IL-17 expression were significantly downtrend [F =3.151,P =0.032,F =4.132,P =0.010].Conclusion The levels of Th17 cells and IL-17 in the peripheral blood of patients with NSCLC significantly correlated with the pathological types and TNM stage,might play an important role in the course of the disease.

Objective To evaluate if the difference between oversea predicted values and healthy adult measurements of spirometry in shenyang.Methods 287 healthy adult volunteers underwent pulmonary function test.The measurements of spiromenty of 230 selected objects were compared to conventional oversea predicted values.Results In male group(154 objects),the measured values were as follows:VC(4.4?0.7)L,FVC(4.3?0.7)L,FEV1(3.5?0.6)L,FEF25%～75%(3.6?1.1)L/s,PEF(7.1?1.9)L/s,IC(2.8?0.7)L,respectively.In female group(76 objects),the measured values were as follows:VC(3.1?0.7)L,FVC(3.1?0.7)L,FEV1 2.5?0.6(L),FEF25%～75%(2.6?0.9)L/s,PEF(5.0?1.4)L/s,IC(2.1?0.7)L respectively.The measurements were 2%～26% and 2%～21% lower than the predicted values in male and female respectively.The major compared values had significant difference.Conclusion The major measured values of spirometry in healthy adult in shenyang were significantly lower than the oversea predicted values.Knudson or ECCS regression equations with appropriate correction factor were suggested to use in shenyang.

Purpose To investigate the diagnosis and differentail diagnosis of metastatic small cell carcinoma of bone in needle biopsy, especially for the differentail diagnosis with Ewing sarcoma of bone. Methods Clinicopathological informations of 11 cases of metastat-ic small cell carcinoma and 20 cases of Ewing sarcoma were collected, and markers for differentail diagnosis were detected in two groups by immunohistochemistry of EnVision. Results The positive rates of CD99 and FLI-1 were 27. 3% and 54. 5% in metastatic small cell carcinoma group, while the positive rate of CK was 15. 0% in Ewing sarcoma group. Patient′s age, single lesion, expression of CK, vimentin, CD99, FLI-1, CD56 were significantly different in two groups. Conclusions Metastatic small cell carcinoma and E-wing sarcoma share similar histopathologic features in needle biopsy, no single immunohistochemical marker can specifically distinguish small cell carcinoma from Ewing sarcouma. The correct diagnosis should comprehensive analyze clinicopathologic characters and a se-ries of immunohistochemical markers.

Objective To verify the feasibility of Cas9 microinjection in frozen-thawed pronuclear embryos, based on the model of pronuclear embryos of C57BL/6J mice by in vitro fertilization.Methods After fertilized mouse pronuclear embryos cultured in vitro, one-cell and 2-cell embryos were frozen using EFS20/40 cryopreservation tube.The next day recovered and then cultured.The recovery rate and survival rate of the one-cell and 2-cell embryos were compared.The frozen-thawed and fresh pronuclear embryos were injected with Cas9 mixture and injection buffer into the cytoplasm, and then cultivated to 2-cell embryos,and the survival rate and development rate of the 2-cell embryos were compared.Results The recovery rate of frozen-thawed one-cell embryos was 92.5%, the survival rate was 92.8%, the recovery rate of 2-cell embryos was 90.5% and the survival rate was 95.8%, showing no significant difference.Furthermore, the survival rate of fresh one-cell embryos after Cas9 injection was 92.7%, the survival rate of one-cell embryos of the blank group was 97.5%.While the survival rate of Cas9 injected frozen-thawed one-cell embryos was 82.6%, and that of the blank group was 92%,showing a significant difference between the frozen-thawed injected group and other groups(P < 0.05).The development rate of 2-cell embryos after Cas9 injection was not significantly different.Conclusions Frozen-thawed pronuclear embryos can be used for Cas9 microinjection.