AIM To explain the variation of blood vessel factors on pulmonary hypertension(PH) induced by monocrotaline(MCT). METHODS Rats were given an injection of MCT ih(60 mg?kg -1 ). The concentration of endothe line(ET), nitric oxide(NO), thromboxane B 2(TXB 2) and 6 keto prostaglandin F 1? (6 keto PGF 1? ) were measured by radioimmunoassay and colorimetric analysis after the model had been set up successfully for 4 wk. RESULTS MCT either raised 6 keto PGF 1? and NO or prevented the progression of ET and TXB 2. CONCLUSION The mechanism of PH induced by MCT probably associated with the variation of those blood vessel factors.

Objective:To observe the effect of photodynamic therapy(PDT)on apoptosis of human lung adenocarcinoma cell line A549.Methods:Human lung adenocarcinoma cell line A549was used and the optimal parameters of PDT were se-lected by MTT colorometric assay.The experiment was divided into2groups:control group and PDT group.Twenty-four hours after PDT(HPD concentration10mg/L,laser dosage10J/cm 2 ),cell cycle and apoptosis rate were estimated by flow cytometry;agarose gel electrophoresis and TUNEL were used to observe the DNA fragments and apoptosis index(AI).Re-sults:The killing effect was saturated when HPD was10mg/L and laser was10J/cm 2 .Under the same laser(10J/cm 2 ),A549cells were exposed to6different duty time fabrications and the outcomes showed that their D 492 had no statistical differ-ence.Agarose gel electrophoresis of DNA showed DNA ladder,a biochemical hallmark of apoptosis in the PDT group.Flow cytometry showed that PDT group had G 0 /G 1 stage arrest.The apoptosis rate was(18.443?7.122)%in PDT group vs (0.301?0.361)% in control group(P

Objective To investigate the changes and significance of plasma interleukin (IL)-6,IL-8,tumor necrosis factor-alpha (TNF-α) and nitric oxide synthase (NOS) in acute blood loss patients.Methods The levels of plasma IL-6,IL-8,TNF-α and NOS were determined in 25 patients with acute blood loss(blood loss group) and 25 healthy controls(control group) by enzyme linked immunosorbent assay.Result The levels of plasma IL-6,IL-8,TNF-α and NOS in blood loss group were higher than those in control group (0.284 ± 0.027 vs.0.204 ± 0.016,0.059 ± 0.079 vs.0.037 ± 0.039,0.460 ± 0.024 vs.0.372 ±0.018,0.637 ±0.054 vs.0.443 ±0.040,P＜0.05 or ＜0.01).Conclusions First,the expressions of IL-6,IL-8,TNF-α and NOS are strongly induced at the circumstances of acute early stage blood loss which can cause inflammatory reaction,aggravate the damage of soft tissues and organs,and easy to lead the blood loss to uncontrolled hemorrhagic shock.Second,the expression of NOS is strongly induced at the circumstances of acute blood loss which should reduce the damage of soft tissue,and alleviate the hemorrhagic shock.Therefore,the levels of plasma IL-6,IL-8,TNF-α and NOS can estimate the illness prognosis and curative effect,which have important clinical instruction value for effective treatment of hemorrhagic shock.

AIM: To establish HPLC fingerprint for the identification of Hydrocotyle sibthorpioides and its adulterants. METHODS: The chromatographic seperation was performed on a Diamonsil C_(18) (250 mm×4.6 mm,5 μm)with a mobile phase consisting of acetonitrile-0.1% phosphoric acid, gradient eluent, at the flow rate of 1.0 mL/min. The UV detection was set at 340 nm. RESULTS: There was an apparent difference in fingerprint between Hydrocotyle sibthorpioides and its adulterants. CONCLUSION: The method is stable and reliable with a good reproducibility andprovides a reference standard for identifying Hydrocotyle sibthorpioides and its adulterants.

Objective To analyze the features of cutaneous lesions and age of onset of tuberous sclerosis(TS).Methods A total of 54 TS patients were recruited;their clinical data were analysed.Results The occurance of facial angiofibromas,periungual fibromas,hypomelanotic macules and shark macules was 72.5%,26%,70%and 48%,respectively,and the median age of onset of these lesions was 7,14,0.2 and 6 years,respectively,with a peak at 1-9,10-19,＜1 and 1-9 years,respectively.Conclusions Cutaneous lesions are commonly seen in TS,and most of them develop early in life.

Objective To investigate the effects and probable mechanism ofShenfu injection on the oxidaitve damage of H2O2-induced PC12 cells.Methods PC12 cells were cultured and exposed to 100μmol/L H2O2 for 1 h to establish the oxidative damage model. The protective effect ofShenfu injection was observed by the cell survival rate measured by colorimetric MTT assay, and the leakage rate of lactic dehydrogense (LDH). Western blot methods were used to detect the expression of NF-κB signaling pathway.Results Compared with the model group,Shenfu injection at 5, 10, 20 ml/L could improve the PC12 cells survival rate (83.11% ± 2.59 %, 87.99% ± 0.59%, 85.26% ± 1.07%vs. 73.82% ± 1.82%;P<0.01 orP<0.05), decrease the LDH leakage rate (32.75% ± 4.10%, 28.52% ± 1.14%, 35.79% ± 1.62%vs. 64.34% ± 3.18%;P<0.01 or P<0.05). Western blot results showed thatShenfu injection could protect the PC12 cells from oxidaitve damage by suppressing the p-p65/p65 (1.30 ± 0.10, 1.17 ± 0.06, 1.37 ± 0.15 vs. 1.70 ± 0.10;P<0.01 orP<0.05), p-IκBα/IκBα (1.07 ± 0.12, 1.00 ± 0.10, 1.03 ± 0.06 vs. 1.17 ± 0.06; P<0.01 orP<0.05).ConclusionShenfu injection has a obvious antioxidant effect on PC12 cells in vitro.

Based on previous report that the Chinese herb Ligustrum lucidum (LL) extract directly inhibited hepatitis C virus (HCV) replicase (NS5B) activity, the active components of LL extract to inhibit HCV NS5B activity and their inhibition mode were investigated in this study. LL extract was separated using ethyl acetate and thin layer chromatography (TLC). The inhibitory activity of separated fractions on HCV NS5B was analyzed by the inhibitory assay of NS5B activity. The results showed that only fractions 1 and 2 inhibited NS5B activity, and fraction 2 possessed higher inhibitory activity than fraction 1. HPLC analysis combined with inhibitory assays indicated that ursolic acid and oleanolic acid are the active components within fractions 1 and 2 to inhibit NS5B activity, separately. Moreover, oleanolic acid possessed higher inhibitory activity than ursolic acid. Further inhibition mode analysis found that both oleanolic acid and ursolic acid suppressed NS5B activity as noncompetitive inhibitors. The Ki values of ursolic acid and oleanolic acid were about 4.7 microg x mL(-1) (10 micromol x kg(-1)) and 2.5 microg x mL(-1) (5.5 micromol x kg(-1)), respectively. Taken together, these results demonstrated that oleanolic acid and ursolic acid suppressed NS5B activity as noncompetitive inhibitors, implying that the two natural products have potential value for HCV therapy.

AIM: To establish HPLC fingerprint for the identification of Hydrocotyle sibthorpioides and its adulterants. METHODS: The chromatographic seperation was performed on a Diamonsil C_18(250 mm?4.6 mm,5 ?m)with a mobile phase consisting of acetonitrile-0.1% phosphoric acid,gradient eluent,at the flow rate of 1.0 mL/min.The UV detection was set at 340 nm. RESULTS: There was an apparent difference in fingerprint between Hydrocotyle sibthorpioides and its adulterants. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for identifying Hydrocotyle sibthorpioides and its adulterants.

Objective To develop a multiplex touchdown PCR for simultaneous detection of extended-spectrum β-lactamases (ESBLs )-producing Enterobacteriaceae and methicillin-resistant Staphylococcus aureus (MRSA ). Methods Blood culture positive specimens from 102 hospitalized patients were collected between March 2013 and September 2014,four pairs of specific primers were designed based on SHV,TEM,and OXA genes of ESBLs-pro-ducing Enterobacteriaceae and MecA gene of MRSA,drug-resistant genes were amplified with single touchdown PCR and multiplex touchdown PCR, the results were compared with Kirby-Bauer disk diffusion method. Results Each single PCR amplified a specific band,four drug-resistant genes were also detected by multiplex touchdown PCR;the lower detection limits of multiplex touchdown PCR for DNA of MecA,SHV,TEM,and OXA were 4.37 ng,2.19 ng,4.53 ng,and 3.59 ng,respectively.Compared with Kirby-Bauer disk diffusion method, the overall sensitivity and specificity of multiplex touchdown PCR were 100.00% and 88.24% respectively,for ES-BLs were 100.00% and 87.23% respectively,for MRSA were both 100.00%.Conclusion A higher sensitivity and specificity multiple touchdown PCR assay has been developed,and it can be used in the rapid diagnosis and epidemi-ology investigation of bloodstream infection caused by ESBLs-producing Enterobacteriaceae and MRSA,and is help-ful for guiding antimicrobial use in clinic.

Aim To investigate the effect of indirubin derivative PHⅡ-7 and TRAIL on proliferation in breast cancer cell MCF-7 and its MDR counterpart MCF-7/ADR and the mechanism.Methods Growth inhibition rate was examined respectively by MTT assay under treatment with TRAIL or PHⅡ-7 or in combination. Cell apoptosis and ROS production were examined by flow cytometry.The change of TRAIL receptors(DR4/DR5 )in mRNA was analysed by realtime PCR.Re-sults IC50 of PHⅡ-7 on MCF-7 and MCF-7/ADR was (4.49 ±1.55 ),(3.44 ±0.90 )μmol · L-1 respec-tively;MDA-MB-231 was TRAIL sensitive cell line, and apparently TRAIL induced apoptosis in MDA-MB-23 1 .Low concentration of PHⅡ-7 in combination with TRAIL could augment TRAIL-induced cytotoxic effect including apoptosis while TRAIL or PHⅡ-7 treatment alone had limited cytotoxity to those cells.Besides, PHⅡ-7 at this concentration had little toxicity to hu-man peripheral blood mononuclear cells even if in com-bination with TRAIL.PHⅡ-7 generated ROS produc-tion inside MCF-7 and MCF-7/ADR cells and up-regu-lated DR4/DR5 expression concentration dependently. Once upon ROS scavenger NAC involved,the effect of TRAIL receptors up-regualtion by expression was abro-gated.Conclusions PHⅡ-7 at low concentration could improve the sensitivities of breast cancer cell MCF-7 and MCF-7/ADR to TRAIL,the mechanism of which may be the ability of ROS production by PHⅡ-7 help up-regulated TRAIL receptor DR4,DR5 .Our re-search set a solid foundation for PHⅡ-7 in combination with TRAIL in future clinical application.