Synchronization isolation method for multiple types of cells from mouse liver / 中华肝脏病杂志

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.

Effects of hepatitis B virus on Th17, Treg and Th17/Treg ratio in different alanine aminetransferase stages / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the effects of hepatitis B virus (HBV) on helper T lymphocytes 17 (Th17), regulatory T lymphocyte (Treg) and Th17/Treg ratio in chronic hepatitis B patients in different alanine aminetransferase (ALT) stages.@*METHODS@#In the study, 336 chronic hepatitis B patients in the first hospital of Lanzhou University were analyzed. The hepatitis B antigen antibody parameters were measured by chemiluminescence immunoassay analyzer, the liver function parameters were measured by automatic biochemical analyzer, the HBV loads were measured by quantitative PCR, Th17, Treg and Th17/Treg ratios were detected by flow cytometry. Among them, 111 cases (ALT < 40 U/L) of ALT were normal hepatitis B, 108 cases of chronic hepatitis B with ALT above normal upper limit and < 2 times higher (40 U/L≤ALT < 80 U/L), and 117 cases of chronic hepatitis B with ALT above 2 times normal upper limit (80 U/L≤ALT). According to the viral load, they were divided into low replication group with HBV DNA < 4.0 lg copies/mL, medium replication group with 4.0 lg copies/mL≤HBV DNA < 6.0 lg copies/mL and high replication group with HBV DNA ≥ 6.0 lg copies / mL. Dunnett T3 variance analysis were used to analyze the effects of HBV on Th17, Treg and Th17/Treg ratio in the chronic hepatitis B patients in different ALT stages. The changes of virological and immunological indexes before and after treatment were observed for 24 weeks of antiviral therapy in the hepatitis B patients with ALT≥double upper limit of normal group.@*RESULTS@#In the ALT normal group, different virus load HBV had minor effects on Th17, Treg and Th17/Treg ratio. In the ALT≥2 times upper limit of normal group, with the virus load increased, Th17 (3.18%±0.79% in low replication group, 3.78%±0.92% in medium replication group and 4.57%±1.15% in high replication group), Treg cells (5.52%±1.58% in low replication group, 5.89%±1.84% in medium replication group and 6.37%±2.35% in high replication group) and their ratio Th17/Treg (0.57±0.25 in low replication group, 0.65±0.29 in medium replication group and 0.73±0.36 in high replication group) were significantly increased (P < 0.05). After entecavir treatment 24 weeks, the patient' s HBV-DNA decreased significantly, Th17 (3.89%±1.02% vs. 2.06%±0.46%), Treg (6.02%±2.03% vs. 5.06%±1.25%), Th17/Treg ratio (0.65±0.28 vs. 0.41±0.14) decreased significantly (P < 0.05).@*CONCLUSION@#Investigation on the effects of HBV on Th17 and Treg cells and their ratios in different ALT states can clarify the effects of HBV on the body from the immunological perspective and can further understand the ALT grouping for antiviral treatment theoretical significance, which is helpful for clinical treatment.

Masseter nerve-innervated free gracilis muscle transfer for smile reanimation in adults / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the outcomes of masseter nerve-innervated free gracilis muscle transfer for smile reanimation in adults and to explore surgical indications. Methods: A retrospective chart review of 37 patients (11 males, 26 females, with (40.3±12.7) years old)who underwent masseter nerve-innervated free gracilis muscle transfer for smile reanimation between 2016 and 2017 was performed. Patient-reported quality of life (facial clinimetric evaluation, FaCE), physician-reported facial function (eFACE facial grading scale), and oral commissure excursion were compared preoperatively and postoperatively. SPSS 11.0 software was used to analyze the data. Results: Thirty-seven patients were followed up in one year after surgery. Significant postoperative improvements were demonstrated for commissure excursion with smile (preoperatively: (-1.2±3.1) mm, postoperatively: (6.1±3.5) mm, t=-31.1, P<0.01), ipsilateral FaCE (preoperatively: (29.4±14.1), postoperatively: (57.6±23.4), t=-38.1, P<0.01), eFACE score (Composite score t=-35.8, Static score t=-29.1, Dynamic score t=-41.3,Midface score t=-43.9, all P<0.01), respectively. Conclusion: Masseter nerve-innervated free gracilis muscle transfer is an good option for dynamic smile reanimation in adult patients who have undergone treatment for long-standing facial paralysis.

Polydatin attenuates airway inflammation in asthmatic mouse model via p38 MAPK/Nrf2/HO-1 pathway / 中国药理学通报

Aim To investigate whether polydatin re-duces airway inflammation in asthmatic mouse model and explore whether this pathway is related to p38 MAPK/Nrf2/HO-1 . Methods After the establish-ment of the OVA-induced asthmatic mouse model, the animals were injected with 30 mg·kg-1 and 45 mg· kg-1 of polydatin diluted in 0. 2 mL normal saline, while the control group was replaced by normal saline. HE, PAS and Masson staining were used to observe the pathological changes of lung tissue. Diff-Quick staining was used to classify and count the number of inflamma-tory cells in BALF. ELISA was used to detect IgE ex-pressions in BALF. The content of ROS in BALF cells was detected by DHR-123 . The activities of antioxidant enzymes SOD, CAT and MDA in BALF were detected by the enzyme-linked immunosorbent assay kit. The expression of HO-1 in lung tissue was detected by im-munohistochemistry. The protein and mRNA expres-sions of Nrf2 and HO-1 in lung tissue of mice were de-tected by Western blot and RT-PCR. Results Poly-datin treatment significantly reduced inflammatory cell infiltration mucosal secretion, goblet cell proliferation and collagen deposition in the lung tissue of mice, and decreased the number of inflammatory cells and the ex-pression of total IgE and ROS in BALF. It also in-creased the levels of antioxidant enzymes such as SOD and CAT, and lowered the level of MDA. Polydatin re-duced the phosphorylation of p38 MAPK in the lung tissue of mice, enhanced the levels of mRNA and pro-tein expressions of Nrf2 and HO-1 and promoted the nuclear transfer of Nrf2 . The above effects of polydatin were dose-dependent. Conclusions Polydatin exerts anti-oxidative effects in OVA-induced asthmatic mouse model via anti-oxidant pathway. The mechanism may be achieved through the p38 MAPK/Nrf2/HO-1 path-way.

The long-term efficacy of PEG-IFN in treatment of chronic hepatitis C with IL28B rs12979860 C/C genotype / 西安交通大学学报(医学版)

Objective To investigate the long-term efficacy of PEG-IFN alpha-2a (PEG-IFNα-2a)plus ribavirin (RBV)in treatment of chronic hepatitis C (CHC)patients with IL28B single nucleotide polymorphisms (SNPs)rs12979860 C/C type in different HCV genotypes.Methods A prospective study was conducted on 38 CHC patients from our hospital's Infection Department from March 2011 to September 2015.The patients were treated with PEG-IFNα-2a/RBV for 48 weeks.A 42-month follow-up of patients was performed after withdrawal of treatment.The main paramenters to value the efficacy were liver function,blood lipids,and sustained virological response (SVR).Results In the CHC patients with IL28B SNP rs12979860 C/C type,the rate of SVR in patients with antiviral therapy had no significant difference between groups 1b and 2a (73.33% and 95.65%,respectively, P＞0.05).After anti-HCV therapy,liver function indices such as ALT,AST,TBIL,TC,TG and HDL all significantly improved in the two groups (all P＜0.05).However,there was no difference in biochemical indices (ALT,GGT,bilirubin,blood lipids)between the two groups (all P＞0.05).Conclusion In CHC patients with IL28B SNP rs12979860 C/C type,the long-term efficacy of PEG-IFNα-2a/RBV is good.IFN-based antiviral therapy has a higher SVR rate,and liver function and lipid metabolism can be significantly improved.

Risk factors of ISUP Modified Gleason score upgrading after radical prostatectomy / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the factors upgrading the International Society of Urological Pathology (ISUP) Gleason score using the specimens from preoperative prostatic biopsy and radical prostatectomy.</p><p><b>METHODS</b>A total of 164 patients diagnosed with prostate cancer by biopsy underwent radical prostatectomy. We retrospectively analyzed their age, prostate volume, preoperative PSA level, PSA density (PSAD) , the time interval between biopsy and surgery, the number of positive punctures, positive surgical margin, seminal vesicle invasion, lymphatic invasion, and Gleason scores from biopsy and prostatectomy. We also determined the predictors of Gleason score upgrading by logistic regression analysis.</p><p><b>RESULTS</b>Of the 164 cases analyzed, 95 (57.93% ) showed a consistency between the Gleason score of preoperative prostatic biopsy and that after radical prostatectomy, 55 (33.54% ) increased and 14 (8.52%) decreased after prostatectomy as compared with preoperative biopsy. The prostate volume (P < 0.01) and biopsy score (P < 0.05) were independent predictors of Gleason score upgrading. The risk of Gleason score upgrading was 27 times higher in the patients with the prostate volume ≤ 25 ml and 9 times higher in the 25-40 ml group than in the > 60 ml group (P < 0.05).</p><p><b>CONCLUSION</b>Low Gleason score of biopsy (≤ 6) and small prostate volume (≤ 40 ml) may be the predictors of Gleason score upgrading after radical prostatectomy.</p>

Fluorosis of coal burning affects the male reproductive system / 中华男科学杂志

Fluorosis of coal burning is a new type of endemic fluorosis in China, which affects the male reproductive system. Furthermore, the content of fluoride in the semen, sperm mortality, sperm concentration and the incidence of infertility are higher in severe fluorosis areas than in mild- and non-fluorosis areas, so are the levels of serum follicle-stimulating hormone and luteinizing hormone. However, the levels of inhibin B, serum testosterone and estradiol show different degrees of reduction in severe fluorosis areas. Accordingly, fluorosis of coal burning, just like other endemic fluorosis, may affect the structure of male reproductive organs, the generation of sperm and reproductive endocrinology, resulting in the decline of men's reproductive ability.

Virtual imaging anatomy of transnasal approach to the clivus / 解放军医学杂志

Objective To observe the virtual anatomy via transnasal approach in a virtual-reality (VR) environment, and to establish a virtual anatomical model of clival region and explore the application value of the virtual-reality technology. Methods Twenty patients (males 11, females 9, aged from 25 to 70 years old with a mean of 43±0.3 years) with no pathological changes in the sellar area and clivus underwent CT angiography with Discovery Ultra 16 and 3.0T MRI thin-slice scan. The data were collected and inputted into the Dextroscope in the DICOM format. Virtual observation of anatomy via transnasal approach and three-dimensional reconstruction of clivus were carried out in the VR environment. Results Virtual reconstruction and measurements of clivus were successfully performed in the 20 patients. The simulation of transnasal approach was also carried out in the VR environment. The lateral side of clival bone removal through transnasal approach was limited by internal carotid artery, the superior limit was sellar floor and the lower clivus could not be revealed completely because of the restriction of the maxilla and hard palate. Conclusions Virtual reconstruction of the clivus and simulation of transnasal approach can be performed exactly by VR technology, which provides imaginal and anatomical basis in dealing with clivus lesions via transnasal approach.

Association of transforming growth factor-β1 gene +869T/C polymorphism with autoimmune thyroid disease / 中国地方病学杂志

Objective To clarify whether the +869T/C polymorphism in the transforming growth factor-β1 (TGF-β1) gene is associated with TGF-β1 expression,and involved in the severity of Graves disease(GD) and Hashimoto's thyroiditis(HT).Methods The TGF-β1+869T/C polymorphism was genotyped by using PCR-sequence specific primers(PCR-SSP) in genomic DNA samples in blood from 158 patients with HT who developed hypothyroidism before they were 45 years old (severe HT) and 125 untreated,euthyroid patients with HT who were older than 45(mild HT).Using the same method,129 euthyroid patients with GD who had been under treatment and were still positive for anti-thyrotropin receptor antibodies (intractable GD) and 130 euthyroid patients with GD in remission and 144 healthy controls were examined.Results It had no difference between GD,HT groups and control group (x2 =1.488,0.439; 0.626,0.005; all P ＞ 0.05 ).The frequency of the TT genotype and the T allele were higher in group with severe HT[34.81％(55/158),58.86％( 186/316)] than in those with mild HT[ 17.60％ (22/125),43.60％ (109/250); x2 =14.040,13.026,all P ＜ 0.05].In contrast,the frequency of the CC genotype was higher in group with intractable GD[ (21.03％(31/129),51.16％(132/258)] than in group with GD in remission[ 13.85％ (18/130),40.38％( 105/260); x2 =12.225,6.061,all P ＜ 0.05 ].TGF-β1 +869 T/C genotype had the correlation with severe groups of HT and GD.C allele would increase in severity of GD(OR =1.546,95％ CI =0.192 - 2.190),and T allele would increase in severity of HT(OR =1.851,95％ CI =1.323 - 2.589).Conclusion The +869T/C polymorphism in the TGF-β1 gene is associated with the severity and intractability of autoimmune thyroid disease.

Transfection of bone marrow mesenchymal stem cells by adeno-associated virus 2/1 vector / 中国实验血液学杂志

This study was aimed to investigate the transfection efficacy of recombinant adeno-associated virus 2/1 (rAAV2/1) on bone marrow mesenchymal stem cells (BMMSCs) at different multiplicities of infection (MOI) and time, and effect of transfection on growth of rat BMMSCs. The rat BMMSCs cultured in vitro were transfected by using rAAV2/1 with enhanced green fluorescent protein (rAAV2/1-EGFP) at MOI of 1 x 10(4), 1 x 10(5) and 1 x 10(6); the EGFP expression was observed by fluorescent microscopy at 3, 7 and 14 days. The viability, proliferation multiple, differentiation ability of daughter cells were detected for evaluating the effect of rAAV2/1 on survival, proliferation and differentiation of BMMSCs and the fluorescence index (FI) were determined by flow cytometry. The results indicated that after transfection with rAAV2/1 for 24 hours the green fluorescence in BMMSCs were observed, but also the fluorescence gradually was enhanced along with prolonging of time, and reached to steady level after 7 days; the viability, proliferation multiple, differentiation ability of BMMSCs transfected by rAAV2/1-EGFP at different MOI showed no significant changes at 3,7 and 14 days (p > 0.05), meanwhile at same MOI the proliferation multiple obviously increased in comparison between 7 day vs 3 day and 14 days vs 7 days (p < 0.01). The flow cytometric detection showed that the transfection efficacy of rAAV2/1-EGFP on BMMSCs and FI increased significantly as the multiplicity of infection and culture time increased (p < 0.05). It is concluded that rAAV2/1-EGFP is able to transfect into BMMSCs effectively, but the transfection efficiency and fluorescence index increase significantly along with increase of multiplicity of infection and culture time. rAAV2/1-EGFP do not affect viability, proliferation multiple and differentiation ability of BMMSCs. rAAV2/1 is a kind of active vector for gene transfer to reform BMMSCs.

Relationships of Serum Leptin,Tumor Necrosis Factor-? and Insulin Resistance in Simple Obesity Children / 实用儿科临床杂志

Objective To explore the relationships of serum leptin(LP),tumor necrosis factor-?(TNF-?)and insulin resistance in simple obesity children.Methods Forty-seven children with simple obesity were divided into two groups: high insulin group(HIG) and normal insulin group(NIG) according to the levels of fasting state of serum insulin and glucose.The fasting state of serum insulin(FINS),LP,TNF-? and fasting blood glucose(FBG) were measured.The relationships of LP,TNF-? and insulin resistance index(HOMA-IR) were also analyzed.Results 1.The BMI,waist,hip,waist hip ratio(WHR),LP,TNF-?,HOMA-IR were obviously higher in the HIG than those in the NIG and NCG groups(Pa0.05).3.In the HIG group,the FINS was positively correlated to LP,TNF-? and HOMA-IR(r=0.560,0.413,0.864 P