OBJECTIVE:To optimize the extraction craft of gentiopicrin from gentian.METHODS:The extraction craft was optimized by orthogonal test with gentiopicrin as an index,and the content of gentiopicroside was determined by TLC scanning method.RESTLUTS:The quantity of menstruum and the extraction times of gentiopicrin had significant influence in the gentiopicrin extraction(P

Objective To investigate the therapeutic effect of epigallocatechin gallate (EGCG) on the bladder neo-plasm induced by N-methyl-N-nitrosourea (MNU) in Wistar rats. Methods Seventy-two female rats were randomly divid-ed into two groups, treatment group and control group. The rat model of bladder tumor was established by intravesical instilla-tion of MNU (2mL/time, once every two weeks) inside bladder. Rats in treatment group were given EGCG (2 mL/time, once/day), and control group were given the same volume of distilled water. Six rats were executed respectively at the 3rd, 5th, 7th, 9th, 11th and 13th week, and the bladder histopathological changes were studied in two groups. Results The untypical hyper-plasia was observed after 5 weeks. The bladder neoplasm was observed at the 11th week in control group. The bladder neo-plasm was not found in treatment group. There was a significantly lower pathological score in treatment group than that of control group except for the first two weeks (P＜0.05). Conclusion EGCG is effective for treatment of the bladder neo-plasm induced by MNU.

AIM:To investigate whether angiotensin-(1-7) [Ang-(1-7)] protects H9c2 cardiac cells against high glucose (HG)-induced injury and inflammation by inhibiting the interaction between Toll-like receptor 4 (TLR4) acti-vation and necroptosis .METHODS:The expression levels of receptor-interacting protein 3 ( RIP3;an indicator of necrop-tosis) and TLR4 were determined by Western blot .Cell viability was measured by CCK-8 assay.The activity of lactate de-hydrogenase ( LDH) in the culture medium was measured with a commercial kit .The releases of interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α) were measured by ELISA .The intracellular level of reactive oxygen species ( ROS) was analyzed by 2 ’ , 7 ’-dichlorfluorescein-diacetate ( DCFH-DA ) stating followed by photofluorography .Mitochondrial membrane potential ( MMP) was examined by rhodamine 123 staining followed by photofluorography .RESULTS:After the H9c2 cardiac cells were treated with HG (35 mmol/L glucose) for 24 h, the expression of RIP3 was obviously increased . Co-treatment of the cells with 30μmol/L TAK-242 (an inhibitor of TLR4) attenuated the up-regulation of RIP3 induced by HG.Furthermore, the expression of TLR4 was significantly increased after the cells were exposed to HG for 24 h, and co-treatment of the cells with 100μmol/L necrostatin-1 ( Nec-1;a specific inhibitor of necroptosis ) and HG for 24 h attenua-ted the up-regulation of TLR4 expression induced by HG .Moreover, 1μmol/L Ang-(1-7) simultaneously blocked the up-regulation of the RIP3 and TLR4 induced by HG.On the other hand, co-treatment of the cells with 1μmol/L Ang-(1-7), 30 μmol/L TAK-242 or 100 μmol/L Nec-1 and HG for 24 h attenuated HG-induced injuries and inflammatory response , leading to the increase in the cell viability , and the decreases in the activity of LDH , ROS generation , MMP loss as well as the releases of IL-1βand TNF-α.CONCLUSION:Ang-(1-7) protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the interaction between TLR 4 activation and necroptosis .

[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.

Objective: To summarize the clinical experience and short-term outcome of minimally invasive occlusion in patients with peri-membranous ventricular septal defect (PmVSD) via right subaxillary route under trans-esophageal echocardiography (TEE) guidance.
Methods: A total of 122 PmVSD patients treated in our hospital from 2014-01 to 2015-07 were summarized. There were 54 male and 68 female with the mean age of (2.7±2.2, 0.5-9.7) years, mean body weight of (13.9±6.0, 6.1-38.0) kg and mean PmVSD diameter of (3.8±0.8, 2.5-7.0) mm. The patients were taken left lateral position, a (2-3) cm incision was performed along right mid-axillary line between the 3rd rib and 4th rib, the thoracic entrance was at 4th inter-costal space. A purse-string suture was conducted on right atrial surface, a special hollow probe was inserted into right atrium and crossed tricuspid into right ventricle under TEE guidance; the probe was adjusted to the point or crossed VSD into left ventricle followed by guide wire insertion to establish a deliver pathway, and ifnally, occlusion device was regularly deployed to close the defect. Post-operative ECG, TEE and chest X-ray were conducted for followed-up study.
Results: There were 119/122 (98.4%) patients occluded successfully and 3 failed patients were converted to cardiopulmonary bypass surgery at the original incision. The average size of occluder was (4.9±1.1, 4-10) mm and all devices were concentric. The patients were followed up at the mean of (8.3±5.0, 1.0-19.8) months, during that period, 12/119 (10.1%) had new mild tricuspid regurgitation, 16 (13.4%) suffered from incomplete right bundle branch block, 4 (3.4%) had small residual shunt and 2 of them were self-closed at 1 and 3 months after operation respectively. There were no complete atrio-ventricular block, no new aortic valve regurgitation and no device dislocation.
Conclusion: Minimally invasive occlusion of PmVSD via right subaxillary route under TEE guidance was a safe, effective, feasible and better cosmetic method for treating relevant patients; while its long-term outcome should be further observed.

Aim To investigate the role of ATP-sensi-tive potassium channels-Akt pathway in exogenous hy-drogen sulfide( H2 S) inhibiting the high glucose( HG)-induced injury in H9c2 cardiac cells. Methods The expression level of Akt protein was tested by Western blot assay. The cell viability was measured by cell counter kit-8(CCK-8 assay). The number of apoptotic cells was tested by Hoechst 33258 nuclear staining fol-lowed by photofluorography. The intracellular levels of reactive oxygen species ( ROS ) were detected by DCFH-DA staining followed by photofluorography. Mi-tochondrial membrane potential ( MMP ) was examined by JC-1 staining followed by photofluorography. Results H9c2 cells were treated with 35 mmol·L-1 glucose (high glucose, HG) for 0 ~24 h respectively. After treating for 3 h, the expression level of phosphorated ( p )-Akt protein began to be obviously reduced, the maximum reduced expression level was observed after the cells were exposed to HG for 24 h. Pretreatment of the cells with 50 μmol · L-1 pinacidil ( Pin, a KATP channel opener) or 400 μmol·L-1 NaHS( a donor of H2 S) prior to exposure to HG considerably blocked the down regulation of p-Akt expression level induced by HG. However, pretreatment with 1 mmol · L-1 KATP channel blocker glibenclamide( Gli) obviously attenua-ted the inhibitory effect of NaHS on HG-induced down-regulation of p-Akt expression level. On the other hand, the protective effects of NaHS against the HG-induced cardiomyocyte injury were markedly blocked by 30 μmol·L-1 Ly294002(an inhibitor of Akt), as indicated by the decrease in cell viability and MMP dissipation as well as the increases in the number of apoptotic cells and ROS generation. Conclution KATP channels-Akt pathway mediates the protective effect of H2 S against the HG-induced cardiac injury.

AIM:Tostudywhe ther theangiotens in-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardio-myocytes against high glucose (HG)-induced injury by inhibiting nuclear factor-κB (NF-κB) pathway.METHODS:The cell viability was measured by CCK-8 assay.The intracellular levels of reactive oxygen species ( ROS) were detected by DCFH-DA staining .The number of apoptotic cells was tested by Hoechst 33258 nuclear staining .Mitochondrial membrane potential ( MMP) was examined by JC-1 staining.The levels of NF-κB p65 subunit and cleaved caspase-3 protein were de-termined by Western blotting.RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose (HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated ( p) NF-κB p65, peaking at 60 min.Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30μmol/L and HG for 24 h inhibited HG-induced cy-totoxicity, evidenced by an increase in cell viability .On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage , indicated by decreases in the number of apoptotic cells , cleaved caspase-3 level, ROS generation and MMP loss .However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor (Mas receptor).Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG.CONCLUSION:Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway .

Aim To investigate whether nicorandil (Nic)protects H9c2 cardiac cells against high glucose (HG)-induced injury and inflammation by inhibiting nuclear factor-κB (NF-κB )/cyclooxygenase-2 (COX-2 )pathway.Methods Cell viability was measured by cell counter kit-8 (CCK-8)assay.The expression lev-els of NF-κB,COX-2 and cleaved caspase-3 were de-termined by Western blot.The activity of lactate dehy-drogenase (LDH)in the culture medium was measured with commercial kits.The intracellular level of reactive oxygen species (ROS)was detected by 2′,7′-dichlor-fluorescein-diacetate (DCFH-DA)staining followed by photofluorography.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography.Mitochondrial membrane poten-tial (MMP)was examined by rhodamine 123 staining followed by photofluorography.The secretion levels of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were detected by ELISA.Results After H9 c2 cardiac cells were treated with 35 mmol · L-1 glucose (high glucose,HG)for 24 h,the cell viability was significantly decreased .Pre-treatment of the cells with 20~100 μmol·L-1 Nic for 60 min or 50 μmol· L-1 Nic for 30~120 min before exposure to HG signif-icantly attenuated the decrease in viability induced by HG.On the other hand,HG increased the expression levels of phosphorated (p)-NF-κB p65 and cyclooxy-genase-2 (COX-2 )in H9c2 cardiac cells.Pre-treat-ment of the cells with 50 μmol·L-1 Nic for 60 min at-tenuated the up-regulation of p-NF-κB p65 and COX-2 expression levels induced by HG.Furthermore,HG induced considerable injuries and inflammatory re-sponse,leading to increases in LDH activity,ROS generation,MMP loss,the number of apoptotic cells, the expression of cleaved caspase-3 as well as the se-cretion levels of IL-1βand TNF-α.Pre-treatment of the cells with 50 μmol·L-1 Nic for 60 min before HG exposure,or co-treatment of the cells with 100 μmol· L-1 PDTC (an inhibitor of NF-κB)or 10 μmol·L-1 NS-398 (an inhibitor of COX-2)and HG for 24 h ob-viously reduced the above injuries and inflammatory re-sponse induced by HG. Conclusion Nic protects H9 c2 cardiac cells against HG-induced injury and in-flammation by inhibiting NF-κB/COX-2 pathway.

Objective: To develop a model for rapid and non-destructive determination of amoxilcillin sodium and sulbactam sodium for injection using the analysis of near infrared diffuse reflectance spectroscopy (NIR) and chemometrics.Methods: Totally 41 batches of commercial samples and 20 batches of laboratory samples were analyzed by NIR and the legal methods.The first derivative and vector normalization were selected as the preprocessing methods and 8 720-5 446 cm-1 was selected as the frequency range.Results: The quantitative model was constructed based on 16 batches of commercial samples and 15 batches of laboratory samples (0.75 g) and the content ranged from 4.45% to 61.82% for amoxilcillin and 15.75% to 30.25% for sulbactam.The root mean square errors of cross validation (RMSECV), determination coefficients (R 2) and root mean square errors of prediction (RMSEP) respectively was 0.858 , 0.998 1 and 0.936 for amoxilcillin, and respectively was 0.541 , 0.988 1 and 0.423 for sulbactam.The model was tested based on 5 batches of commercial samples and 5 batches of laboratory samples (0.75 g) and the results well met with those of the legal methods with difference ≤ 1.5 %.The model also applied in 10 batches of commercial samples (1.5 g) and 2 batches of samples from the other manufacturers.Conclusion: The non-destructive quantitative NIR methods are accurate with good reproducibility, and applicable for the rapid analysis of amoxilcillin sodium and sulbactam sodium for injection.

AIM: To investigate effects of Retinervus luffae fructus (RLF) on level of serum lipid and body weight in hyperlipidemia rats. METHODS: Thirty-two male SD rats were randomly divided into four groups: control group (A), hyperlipidemia group (B), hyperlipidemia + RLF group (C), RLF group (D). Both group A and C were fed normal diet every day, while group B and group D fed high fat diet. Meanwhile, group C and D were administered with RLF solution at the dose of 10 mL/kg, respectively for 14 days, while group A and B were administered with drinking water. RESULTS: (1) At the end of experiment, a significant reduction was found in the levels of serum total cholesterol (TC) and triglyceride (TG) of group C animals treated with RLF solution; (2) The levels of serum TC of group D was progressively decreased compared to the level of serum TC at the beginning of experiment; (3) The level of high-density lipoprotein cholesterol (HDL-C) of group C remained unaltered 8d after treatment with RLF solution; (4) The body weight in group C was obviously lower than that in group B. CONCLUSION: RLF had an obvious hypolipidemic effect on hyperlipidemia rats. It can inhibit the decrease in the HDL-C and the increase of body weight in rats. [