Objective To study the effect of mitochondrial permeability transition(MPT) after ischemia/reperfusion(I/R), and to find the relationship between MPT and apoptosis of hepatocytes in rats. Methods SD rats were divided into three groups:sham operation group, I/R group, and I/R+CsA group. A model of hepatic I/R in rats was made according to the method of Nauta. Active caspase-3 in cytoplasm was examined by immunohistochemistry, and the content of cytochrome C protein was evaluated by Western blotting. The incidence of apoptic hepatocytes was determined by TUNEL method. Cyclosporine A(10 mg?kg -1?d -1) was used as an inhibitor of MPT owing to its inhibitive effect on mitochondrial permeability transition pore(PT pore), and the influences of CsA were analyzed. Results Pretreated with CsA ameliorated injury after reperfusion. The number of apoptotic liver cells was decreased. CsA significantly prevented the release of cytochrome C from mitochondria, demonstrating its effect on MPT through preventing PT pore from opening. Conclusion MPT may play a key role in the apoptosis of liver cell after I/R. CsA, a powerful inhibitor of PT pore, is capable of abating the effect of MPT, reducing the release of cytochrome C, retarding the activation of caspase-3 and hence ameliorates apoptosis of hepatocytes following I/R procedure.

ObjectiveTo explore the value of Tth-single strand binding protein (SSB) used as an additive to improve the polymerase chain reaction (PCR) specificity for single nucleotide polymorphisms(SNP) alleles genotyping.MethodsTth-SSB plasmid was constructed and the protein was expressed,then the expressed Tth-SSB was added into PCR system detecting cytochrome P450 Protein ( CYP2C19 * 3,636G＞A)genotype to determine the optimal usage and condition.Then,the genotypes of 30 cerebral ischemia patients were tested with established methods and compared with direct sequencing to verify the accuracy of Tth-SSB as an additive into PCR for SNP genotyping.ResultsThe purity of Tth-SSB was 85％ and optimal dosage was 1 μg.The protein could improve the specificity and reduce the dimer when Tth-SSB was added into the PCR system.Thirty patients genotyping results as follow:26 patients belong to G/G homozygote,4 patients belong to G/A heterozygote,no body belong to A/A homozygote.The coincidence acquire 100％ with parallel sequencing.ConclusionAs an additive,Tth-SSB could significantly improve the accuracy of genotyping by eliminating non-specific bands.

Objective To explore antibiotics resistance profiles and DNA fingerprints of Pseudomonas aeruginosa isolates resistant to imipenem (IRPA). Methods DNA fingerprints of 56 strains isolated from ICU (intensive care unit) were constructed by ERIC-PCR (enterobacter repetitive intergenic consensus-PCR). MICs (minimal inhibitory concentrations) were determined by agar dilution method.Results 33 genotypes were got from 56 strains by ERIC-PCR. Of 8 frequently used antibiotics, 5 of them showed resistance rate higher than 50%. Conclusion It is high time to pay attention to multi-drug resistance of IRPA. The exist of prevalence of IRPA clone in ICU advise us to control of IRPA in hospital effectively.

Objective To measure the expression of Caspase-3 and P-gp in hepatocellular carcinoma (HCC) and adjacent tissues, and to examine these two proteins' relationships with HCC clinical and pathological characteristics and the associations between the two proteins. Methods The expression of Caspase-3 and P-gp were measured in 50 HCC tissues and adjacent tissues by EnvisionTM immunohistochemistry. HCC clinic and pathological characteristics and follow-up data were also collected and analyzed. Results The expression of Caspase-3 in HCC tissues was significantly lower than that in adjacent tissues(P <0.05). The expression of Caspase-3 in HCC tissues was significantly associated with HCC's Edmonson grade and the presence of hepatitis envelope(P <0.05). On the other hand, the expression of P-gp in HCC tissues was significantly higher than that in adjacent tissues( P <0.05 ). P-gp expression in HCC tissues was significantly correlated with HCC's Edmonson grade and the presence of cirrhosis(P < 0.05 ). The expression of Caspase-3 and P-gp was negatively correlated ( r = - 0. 600,P = 0.000). A better prognosis was observed among HCC patients with either positive Caspase-3 expression or negative P-gp expression(P < 0.05). Conclusion Caspase-3 may promote apoptosis and inhibit the development of HCC, while P-gp may have an anti-apoptosis function and play a role in HCC's drug resistance. P-gp can inhibit the function of Caspase-3 and an absence or low expression of Caspase-3 may be related to apoptosis resistance and multidrug resistance.

Eight 1,6-O,O-diacetylbritannilactone (OABL) derivatives (compounds 1-8) were synthesized by esterification or reduction of 1-O-diacetylbritannilactone (ABL) isolated from Inula japonica.All derivatives were evaluated for their anti-inflammation activities through the determination of inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced macrophages.As results,compounds 5-8 (IC50 ＜ 2 μmol/L) exhibited more potent inhibition of NO production activities than the lead compound OABL.

Objective:To explore the clinical regularities in acupuncture-moxibustion treatment of cancer pain by reviewing the relevant studies published between 1985 and 2017.Methods:Based on the Chinese Medicine Acupuncture-moxibustion Information Database,quantitative analysis,correlation analysis and Chi-square test were applied to analyze the commonly used acupoints,meridian affiliations and body region distributions,commonly used methods and acupoint correlations,treatment method correlations,the efficacy of acupuncture-moxibustion plus the three-step analgesic ladder for cancer pain,and indicators.Results:Zusanli (ST 36),Ashi point and Sanyinjiao (SP 6) ranked the top on the list of frequency;points from Bladder Meridian of Foot Taiyang and Stomach Meridian of Foot Yangming were often used;points from the lower limbs and back had high frequencies.The most commonly used treatment method was acupuncture-moxibustion plus medication.Acupuncture-moxibustion plus the three-step analgesic ladder showed certain advantage compared with the two methods used separately,and among the integrated methods,acupoint application plus the three-step analgesic ladder produced the most significant efficacy.It took (44.77±55.54) min for the analgesic effect to act and the effect lasted for (12.81±14.59) h.Numerical rating scale (NRS),visual analog scale (VAS) and Karnofsky performance status (KPS) scores all showed significant changes after interventions (all P＜0.01);there was no significant change in the score of quality of life (QOL) after interventions (P＞0.05).Conclusion:Zusanli (ST 36),Ashi point and Sanyinjiao (SP 6) are commonly selected in acupuncture-moxibustion treatment of cancer pain;acupuncture and acupoint application are often used;acupuncture-moxibustion plus the three-step analgesic ladder can boost the treatment efficacy.

Objective To evaluate the using of either 225 or 150 microgrammes of mifepristone combined with misoprostol for termination of second-trimester gestation(16～24 weeks).Methods 180 women requesting voluntary induced abortion during gestation 16～24 weeks were randomised to three groups,group 1:oral mifepris- tone 225rag,group 2:oral mifepristone 150mg,and group 3:injected 100rag rivanot by amniocentestis.The total suc- cess rate,once success rate,the interval of having-medicine to uterine-constraction,the volume of bleeding within 2 hours after labour and cervical laceration rate were observed.Results The once success rate of induced labour in group 1 was higher than that in group 2 and group 3(P

Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi's extracts. Methods Twelve Sprague-Dawley rats'right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium,NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi;the normal group contained the extracts of the normal hippocampi;the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs into neurons.

Objective To investigate the relationship between the nerve growth factor ( NGF ) induced hippocampal neuroregeneration and homeobox gene Lhx 8.Methods Seventy-two SD rats were divided into control group , transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection . Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group.The choline acetyltransferase ( ChAT)/Lhx8 double labeled cells in subgranular zone ( SGZ) of hippocampus in each group were detected by immunofluorescence .Results The expression of Lhx8 gene and protein in the transected , NGF group and especially in the transected combined with NGF group was obviously higher than in the control group .The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group . Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

?AIM: To study the association of the single nucleotide polymorphism ( SNP) rs1157699 in the calcitonin receptor-like receptor ( CRLR ) gene with primary angle closure ( PAC) in a Han Chinese population.?METHODS: All samples, involved 232 PAC cases and 306 controls, were obtained from an epidemiologic survey conducted in Funing, Jiangsu Province, China. Genotyping were carried out by TaqMan-MGB probe using the real time quantitative polymerase chain reaction system to study the relationship between SNP of rs1157699 in CRLR gene and PAC.?RESULTS: The prevalence of CRLRrs1157699 genotype was 67.4%, 30.0%, 2.6% for CC, CT, TT in cases, and 71.3%, 27.0%, 1.7% in controls respectively.There was no difference between the two groups in the distribution of genotype and allele frequencies of rs1157699 (P>0.05).?CONCLUSION:Our results do not support a significant role for rs1157699 in CRLR with PAC.