Objective To find antitumor cOnstituents from Tripterygium hypoglaucum.Methods The chloroform extract of T.hypoglaucum was separated by silica gel column chromatography and preparative HPLC.The structures of compounds isolated were identified by spectral analysis and chemical evidence.Results Seven compounds were isolated and identified as rhein ethyl ester(1),chrysophenol(2),physcion(3),emodin(4),wilfordine(5),wilforgine(6),and wilforine(7).The cytotoxic activities of the compounds against cancer cell lines were assayed.Conclusion Compound 1 is a new natural compound with strong activities against human cancer cell lines (A2780 and OVCAR-3).Compounds 2-4 are isolated from this genus plants for the first time.The possible structure-activity relationship among compounds 1-4 shows that the methoxy group or oxyethyl moiety might be responsible for the cytotoxity.

Cytochrome P450 enzymes are composed of many isozymes and involved in the biotransformation of both exogenous and endogenous substances. A growing number of studies have found that the P450 enzymes do not always follow the classical Michaelis-Menten kinetics, but show atypical kinetic behavior, which is also the current research hotspot. In this paper, the category and mechanisms of atypical kinetics of the P450 enzyme were reviewed, providing theoretical basis for the research of enzyme kinetics.

Objective To evaluate the clinical value of multislice-CT angiography (MSCTA)in planning for the patients undergoing deep inferior epigastric artery perforator (DIEAP) flap operations. Methods Eighteen patients were performed with a 16-slice CT scanner to evaluate the deep inferior epigastric artery perforator prior to DIEAP flap operations. Axial, multiplanar reconstruction( MPR), maximum intensity projection(MIP) and volume rendered (VR) images were analysed and the origins, calibers, courses and anatomic relationships of the deep inferior epigastric artery perforator were evaluated. The anastomosis between the superficial inferior epigastric artery and the main perforator was observed as well. The images were classified into three grades based on the vessels'appearance. A + indicated the vessel appeared clear,continuous and thick. A- indicated the vessel appeared foggy,discontinuous and thin or the vessel partly showed. B indicated no related vessel can be seen. Other 18 patients undergoing conventional abdomen-pelvis CT scans for other reasons were used for control group to compare their CT findings of the deep inferior epigastric artery perforator. Results MSCTA well showed the course of the deep inferior epigastric artery (DIEA). Of the 18 cases, 17 cases appeared as A +, another one A -. It precisely displayed the origins, subcutaneous and intramuscular courses, relations of the main perforators on all cases of showing A +. The exact points where the chosen perforator vessels emerged from the rectus abdominis muscle fascia were located precisely. The superficial inferior epigastric arteries were mostly displayed and the connection between the arteries and the largest-caliber perforator from the deep system could also be shown clearly. Strict concordance with operative findings was found in CTA. Conclusion MSCTA can precisely locate the chosen perforator vessels emerging from the rectus abdominis muscle fascia and it may be a feasible, fast, safe and effective method for preoperative evaluation of DIEAP.

Objective To observe the effects of electro-acupuncture ( EA ) on learning, memory and the morphology of hippocampal neural tissues in rats with a model of chronic cerebral ischemia.Methods One hundred and twenty male Sprague-Dawley rats were used. Chronic cerebral ischemia models were successfully established in 104 of them, and those rats were randomly divided into an EA group and a model group with 52 rats each. These were further subdivided into 1,2, 4 and 6 week subgroups with 13 rats in each. The EA group was given EA. The changes in spatial learning and memory ability were observed using a Morris water maze. The morphological changes in hippocampal nerve tissue were observed by HE staining.Results The escape latency in the EA group was significantly different from the model group at the 2nd, 4th and 6th week. The nerve cells in the dentate gyrus were more tightly and consistently lined-up and had rich layers, and the structures in the EA group were better than in the model group.Conclusions EA can improve spatial learning and memory and promote the repair of injury after cerebral ischemia.

Objective To explore the the role of fraction exhaled nitric oxide(FeNO) in airway inflammation of Mycoplasma pneumoniae pneumonia(MPP).Methods Inpatients with low respiratory tract infection were enrolled from August to November in 2012,69 patients had MPP and 33 had no MPP(non-MPP).Patients with MPP were further grouped into a bronchopneumonia group and the lobar pneumonia group.Fifty-four inguinal hernia patients without respiratory tract infection during the last 2 weeks were enrolled as a control group.FeNO was measured by nitric oxide analyzer.Eosinophile level was detected by blood cells analysator.Results The level of FeNO in patients with MPP [(6.28 ±3.00) ppb] was lower than that of patients of non-MPP [(10.85 ± 2.86) ppb] and the control group [(9.74 ± 3.10) ppb] (t =7.30,6.26,respectively,all P ＜ 0.000 1) ; the level of FeNO between the bronchopneumonia group [(5.78 ± 3.06) ppb] and the lobar pneumonia group [(6.48 ± 2.98) ppb] with infection of Mycoplasma pneumoniae (MP) had no statistical significance(t =0.88,P ＞0.05).The proportion of blood eosinophile in patients with MPP [(0.60 ±0.51) ％] was lower than that of non-MPP group [(1.15 ± 0.76) ％] (t =4.14,P ＜ 0.000 1) ; the proportion of blood eosinophile between bronchopneumonia group [(0.61 ± 0.57) ％] and lobar pneumonia group [(0.60 ± 0.55) ％] with infection of MP had no discrepance (t =-0.05,P ＞0.05).Conclusions MP infection decreases production of FeNO.The possible mechanism for this phenomenon is that the cilia loss and hyperimmune response to MP may affect the production of FeNO.The airway inflammation of mycoplasma pneumonia is associated with cilia loss and hyperimmune response.

Objective To compare the efficacy and safety of two intense pulsed light (IPL) devices for the treatment of facial photoaging.Methods A randomized split-face clinical trial was conducted,and 30 female subjects with facial photoaging were enrolled and randomized to receive treatment with Lumenis One on one half of the face and BBL on the other facial side,once every 3-5 weeks for 5 sessions.Each subject was followed up before the first treatment (the first interview),4 weeks after the third treatment (the second interview),4 weeks after the fifth treatment (the third interview) and 8 weeks after the fifth treatment (the fourth interview).During each follow-up period,global scores for photoaging (GSP) were used to evaluate the photoaging degree on the whole face,a 4-level grading method was applied to evaluate the improvement degree of 5 photoaging signs on each facial side,including wrinkles,skin texture,pigmented spots,telangiectasia and skin tightening,and the visual analogue scale (VAS) to assess pain induced by treatment.After the last treatment,self-assessment on the degree of satisfaction with therapeutic effects was conducted in subjects.Comparisons in the GSP and improvement scores between the two facial sides were conducted by repeated measures analysis of variance (ANOVA).Results A total of 26 subjects completed all the treatments and follow-up.Evaluation of the whole face showed that the GSP significantly decreased from 3.19 ± 0.75 before the first treatment to 2.15 ± 0.83 at 4 weeks after the third treatment (P ＜ 0.01).At 4 and 8 weeks after the fifth treatment,the GSP decreased to 1.85 ± 0.88 and 1.85 ± 0.97 respectively,and no significant difference was observed between the two GSPs (P ＞ 0.01).Evaluation of each facial side showed that improvement scores of skin texture,pigmented spots,telangiectasia and skin tightening on the two facial sides all increased at first and then decreased over the treatment time (Ftime =18.75,10.25,12.83,15.73,respectively,all P ＜ 0.05),and the improvement scores significantly increased at 4 weeks after the fifth treatment compared with those at 4 weeks after the third treatment (all P ＜ 0.017).There were no significant differences in the improvement scores of skin texture,telangiectasia and skin tightening between the third and the fourth interview,but the improvement score of pigmented spots decreased slightly at 8 weeks after the fifth treatment compared with that at 4 weeks after the fifth treatment (P ＜ 0.017).During the whole treatment period,no evident improvement was observed in wrinkles (Ftime =3.17,P ＞ 0.05),and improvement scores of 5 photoaging signs did not differ between the Lumenis One-treated side and BBL-treated side (all P ＞ 0.05).In addition,the VAS pain score was significantly lower in the BBL-treated side than that in the Lumenis One-treated side (4.62 ± 1.54 vs.5.80 ± 1.74,t =2.87,P ＜ 0.05).Most subjects were satisfied with the therapeutic effects (88.46％,23/26).Conclusion Both Lumenis One and BBL can be applied to treat facial photoaging safely and effectively,and improve signs of photoaging such as pigmented spots and skin texture,but the degree of pain on the BBL-treated side is milder than that on the Lumenis One-treated side.

Objective To elucidate the features of clinicopathology and mutation in Fabry disease complicated with thin basement membrane nephropathy (TBMN), and to investigate the kindred. Methods Data of clinicopathology and gene mutation of a female patient of Fabry disease complicated with TBMN admitted to the Department of Nephro]ogy in our hospital were analyzed. Members of her kindred were investigated simultaneously. Results Proband was a 41-year-old Chinese woman who presented syndrome of Fabry disease and TBMN including angiokeratomas, chronic pain, tinnitus, vertigo, left ventricular hypertrophy and nephropathy as proteinuria, microscopic hematuria and hypertension. A percutaneous renal biopsy was performed on the proband, which was consistent with FSGS and vaculization of podocytes by light microscopy.Electron microscopy showed concentric lamellated inclusions in some podocytes. Diffuse thinning of glomerular basement membrane (GBM) with a mean thickness of (216±31) nm was found. The diagnosis of TBMN with suspected Fabry disease was identified. Family screening showed that her daughter had microscopic hematuria, tinnitus and neuropathic pain. One of her sisters only had microscopic hematuria. The activity of α-galacsidase A (α-Gal A )enzyme in the proband and her daughter was 33 units and 75 units respectively (the normal range is 100 to 500 units). They all carried the novel GLA mutation 1208 ins 21 bp and COL4A3 SNP c: 3627G＞A(p:M1209I). While her sister who only had microscopic hematuria just carried the variant of COL4A3 gene-c:3627G ＞A (p:M1209I), and had the normal activity of α-Gal A with no mutation of GLA.Conclusion TBMN should be considered in the patients of Fabry disease with the condition of benign familial hematuria.

Objective:To transfect Mcl-1shRNA into Raw264.7 cells,and screen out specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene to figure out the effect of shRNA on Mcl-1 expression in murine macrophage cell line Raw 264.7.Methods: Specific shRNA was transfected into murine macrophage cell line Raw 264.7 via lipofectamine.Semi-quantitative RT-PCR and Western blot were respectively employed to test the changes in Mcl-1 mRNA level and Mcl-1 protein expressions 24 h and 48 h after the transfection ,and the silencing effects of the three pairs of specific shRNA fragments corresponding to different sites were analyzed.Results: Specific shRNA fragments at 24 h and 48 h could effectively reduce Mcl-1 mRNA and protein level ,with higher silencing effects than those of the normal group ,the lipofectamine group and the negative control group.There were statistically significant differences among them ( P<0.05 ).Among the three pairs of specific shRNA fragments corre-sponding to different sites ,Mcl-1 shRNA3 showed the most significant inhibiting effect on Mcl-1 mRNA and proteins.Conclusion:RNA interference can downregulate the level of Mcl-1 mRNA in murine macrophage cell line Raw 264.7 and greatly downregulate the expression of Mcl-1protein.Specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene have been screened out successfully.

Objective To study the expressions of myeloid cell leukemia-1 (Mcl-1 ) gene in mouse macrophages Raw264.7 and human macrophages THP-1,to screen out the cell lines with high levels of expression as the experimental cells,and based on the screening results to construct the short hairpin RNA(shRNA)eukaryotic expression plasmid targeting mice Mcl-1 gene for transfection and further screen out the shRNA expression plasmid with the most obvious effect in silencing Mcl-1 gene.Methods Semi-quantitative PCR method was used to detect the expression of Mcl-1 mRNA in the two kinds of macrophages.Western blotting was adopted to detect the expressions of Mcl-1 proteins in the two kinds of macrophages.Three different gene loci for Mcl-1 shRNA fragments were designed with small molecules interfering RNA (siRNA)software.Eukaryotic expression plasmid Mcl-1 shRNA 1-3 carrying the shRNA fragments was constructed by a company.And the eukaryotic expression plasmid vector was transfected into scavenger cells Raw264.7 mice via through the liposome.The transfection results 24 h and 48 h after transfection were observed under the inverted fluorescence microscope;Mcl-1 mRNA and protein expression were detected by real time quantitative PCR and Western blot,respectively.Results The relative expression levels of Mcl-1 mRNA and protein in mouse macrophages Raw264.7 were significantly higher than those of human macrophages (P <0.05).The shRNA expression vector constructed within 24 h and 48 h could decrease Mcl-1 mRNA and protein levels in Raw264.7 cells,especially with the most obvious silencing effect at 48 h.The 48-h transfection group differed significantly from normal group,liposome group and negative control group (P <0.05).Compared with Mcl-1shRNA1and Mcl-1shRNA2,Mcl-1shRNA3 had the strongest effect in silencing Mcl-1mRNA and protein.Conclusion We have successfully screened out experimental Raw264.7 cells and Mcl-1 shRNA eukaryotic expression plasmid which has an obvious silencing effect targeting on Mcl-1 in mice macrophages Raw264.7.

AIM:To investigate the effect of inhibiting Mcl-1 gene expression on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis using a technique of RNA interference .METH-ODS:The BALB/c mice were infected with prepared bacterium of the virulence strains of Xinjiang , H37Rv, H37Ra and BCG.Mcl-1-shRNA was applied to the mouse model of infection , and the control groups were set up .On 1 d, 3 d, 5 d and 7 d, the mouse peritoneal macrophages were collected .The expression of Mcl-1 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The apoptotic rate of peritoneal macrophages was analyzed by flow cytometry . RESULTS:The expression of Mcl-1 at mRNA and protein levels was up-regulated in the peritoneal macrophages from the mice infected with different virulence of Mycobacterium tuberculosis, and the cells from the mice infected with virulence strains of Xinjiang and H37Rv expressed higher level of Mcl-1 than the uninfected control cells (P<0.05).The expres-sion of Mcl-1 at mRNA and protein levels was reduced by RNA interference as compared with control group ( P<0.05 ) . Inhibition of Mcl-1 expression induced apoptosis of peritoneal macrophages in the mice .CONCLUSION: The Mcl-1 ex-pression at mRNA and protein levels in mouse peritoneal macrophages infected with different virulence of Mycobacterium tu-berculosis was effectively suppressed by Mcl-1-shRNA, which can induce macrophage apoptosis .