From the water soluble extract of Flos Lonicera japonica Thunb.,three triterpenoid saponins were isolated. On the basis of detailed analysis of 1HNMR, 13CNMR, 1H-1HCOSY, HMBC, HMQC, FAB MS, their structures were decided as 3-O-?- L-rhamnopyranosyl-(1→2)-?-L-arabinopyranosyl hederagenin 28-O-?-Dxylpyranosyl (1→6 )-?-D-glucopyranosyl ester (Ⅰ), 3-O-?-L-arabinopyranosyl hederagenin 28-O-?-L-rhamnopyranosyl (1→2) [?-D-xylpyranosyl (1→6)] -?-glucopyranosyl ester (Ⅱ) a nd 3-O-?-L-rhamnopyranosyl (1→2) -?-L-arabinopyranosyl hederagenin 28-O-?-L-rhamnopyranosyl (1→2)[?-D-xylpyranosyl (1→6)]?-D-glucopyranosyl ester（Ⅲ), and chlorogenin tetraacetate (Ⅳ). Pharmacological experiment showed thatthese compounds had cytoprotective effects against carbon tetrachloride induced hepatic injury.

Animal experiment on influenza virus infection carries certain biohazard risk, with a threat to the health of researchers and public health.The risk levels differ by influenza virus types and subtypes.This article combs the domestic and national laws and rules, and explores the biosafety management of animal study on influenza virus.

Objective To investigate the effect of donepezil hydrochloride on the expression of Calpain Ⅰ-Cdk5/p25 pathway in the hippocampal CA1 area by cerebral ischemia and reperfusion in mice.Methods Mice were divided into model group,sham-operated group and donepezil-treated group.The expression of Calpain Ⅰ in hippocampal CA1 area was measured by immunohistochemistry staining respectively at 4,6 and 8 weeks post cerebral ischemia and reperfusion.Western blot was used to evaluate Cdk5 and p25 protein expression.Results The abilities of learning and memory performance was damaged significantly at 4,6 and 8 weeks after surgery compared to sham-operated group (P＜ 0.05).The expression of Calpain Ⅰ of model group were (0.098 ± 0.009),(0.129 ±0.01),(0.116 ± 0.01),which were higher than that of sham-operated group (0.03 ± 0.003),(0.031 ± 0.003),(0.029 ±0.003) and there was significant difference (P ＜ 0.05).The expression of Cdk5 in model group was (0.54 ± 0.05),(0.73 ± 0.07),(0.7 ± 0.06),which were higher than that of sham-operated group (0.23 ±0.02),(0.31 ± 0.02),(0.33 ± 0.02) and there was significant difference (P ＜ 0.05).The expression of p25 in model group was (0.44 ± 0.04),(0.51 ± 0.04),(0.55 ± 0.06),which were higher than that of sham-operated group(0.19 ± 0.02),(0.24 ± 0.02),(0.2 ± 0.02) and there was significant difference (P ＜ 0.05).The expression of Calpain Ⅰ of donepezil-treated group was (0.041 ± 0.004),(0.054 ± 0.004),(0.046 ± 0.003),which were lower than that of model group.The expression of Cdk5 was (0.28 ± 0.02),(0.33 ± 0.03),(0.38 ± 0.02),and expression of p25 was (0.26 ± 0.02),(0.25 ± 0.03),(0.21 ± 0.02),which were lower than that of model group respectively(P ＜ 0.05).Conclusion Donepezil hydrochloride probably improve the learning and memory abilities by reducing the expression of Calpain Ⅰ and Cdk5/p25.

ObjectiveTo inyestigate the effect of miRNA-155 expression on the differentiations and functions of CD4 + T lymphocyte in patients with unstable angina pectoris. MethodsTwenty-one patients with unstable angina pectoris (UAP) were enrolled in this study, and another 18 patients with normal coronary arteries evidenced by angiography were assigned as control group. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of miRNA-155 in CD4 + T lymphocyte of the peripheral blood. And the levels of IFN-γ and IL-4 in serum were determined by using enzyme-linked immunosorbent assay (ELISA). The CD4 + T lymphocyte were isolated from mononuclear cells prepared with Ficoll-Hypaque density-gradients centrifugation from human peripheral blood by magnetic cell sorting system. Then, the CD4 +T cells (2 × 106 cells/mL) were seeded in culture plates with 6 wells.The CD4 + T lymphocytes were divided into 3 groups in experiment: control group, miRNA-155 group, and miRNA-155 inhibitor group. The numbers of Th1 and Th2 cells were measured by flow cytometry analysis (FACS).The protein levels and expressions of IFN-γRα, T-bet, GATA-3 mRNA were measured by using western blotting and qRT-PCR, respectively. The levels of IFN-γ and IL-4 in culture supernatants of CD4 +T lymphocytes were detected by using ELISA. ResultsIn comparison with the control group, there was significant increase in the expression of miRNA-155 and serum level of IFN-γ in the UAP group (P ＜0. 01 ). There was a positive correlation between miRNA-155 and serum IFN-γ ( r =0. 842, P ＜ 0. 0l ).However, in vitro, the number of Th1, the protein level and expression of T-bet mRNA, and supernatant IFN-γ increased, and the protein level and expression of IFN-γRα protein decreased in miRNA-155 group in comparison with the control group (all P ＜0. 01 ). Interestingly, there were no significant differences in the number of Th2 cells, the expressions of GATA-3mRNA and IFN-γRα mRNA, GATA-3 protein and supernatant IL-4 between control group and miRNA-155 group ( all P ＞ 0. 05 ). And the miRNA-155 inhibitor could attenuate the effect of miRNA-155 effectively. ConclusionsThe miRNA-155 can promote differentiations and improve the function of Thl, playing an important role in the pathogenesis of unstable angina pectoris.

BACKGROUND:Currently,there are many studies concerning the pathogenesis,process,and damage of glaucoma,however,there is not an ideal glaucoma modelOBJECTIVE:To prepare rabbit intraocular hypertension models using three different material injections,and to verify the practical value of intraocular hypetension modelsMETHODS:Thidy New Zealand rabbits were divided randomly into 3 groups,with 10 animals in each group.One eye of each rabbit was served as the experimental eyes and the other eye as control eyes.Autoblood.methyl cellulose,C3F8 was injected into the anterior chamber of the experimental eyes.and the normal saline was injected into the control eyes.The intraocular pressure(IOP)was monitored prior to injection and at hours 0,24,36,48,72,96.120 and 168 after injection.RESULTS AND CONCLUSION:Intraocular hype Rension models could be induced by injecting 3 kinds of materials,and the IOP was obviously increased after injection(P＜0.05),and the ranges and periods of increasing were varied.The periods of increasing of 3 materials were 1,3 and 7 days,respectively,which could maintain for longer time for a second injection.The IOP ranged 1.86-6.65 kPa,and mild anterior segment inflammation could be found.The experiment demonstrated that intraocular hypeansion models using three different material injections are ideal models,which is characterized by simple,reliable and controllable.The suitable model can be selected for acute or chronic glaucoma research.

The reasons why the relationships between doctors and patients hasn't advanced to a full and true harmonious stage in our country are multiple.From the legal point of view,the research on the important legal issues of doctor-patient relationship is deficient.The law regulating the relationship is hysteretic and even lacking.The system settling the doctor-patient conflict is distempered and imperfect.Both the doctors' and patients' legal knowledge and consciousness are inadequate.Therefore,it is necessary to discuss the legal measures from the four aspects to construct a harmonious medical service system.

Objective To observe the influences of NaAsO2 on H3K36me3 modifications,mRNA transcription of O6-methylguanine-DNA methyltransferase gene (MGMT) in HaCaT cells,and to explore the relationship between the transcription of MGMT gene regulated by H3K36me3 and DNA damage induced by arsenic,in order to provide new ideas and scientific basis for prevention and intervention of arsenism.Methods HaCaT cells were treated with 1.25,2.50,5.00 and 10.00 μmol/L NaAsO2 for 24 h,and were also treated with 10.00 μmol/L NaAsO2 for 6,12 and 24 h.HaCaT cells that treated with 0.00 pmol/L NaAsO2 and 0 h were used as blank control group.The degree of DNA damage in peripheral blood cells was detected by single cell gel electrophoresis.The level of H3K36me3 modifications was detected using Western blotting.Quantitative real-time polymerase chain reaction was used to detect the mRNA levels of MGMT gene.Quantitative chromatin immuno-precipitation was used to detect the level of H3K36me3 modifications in the coding regions (ChIP1 and ChIP2) of MGMT gene.Results ①Among the groups of HaCaT cells treated with 2.50,5.00 and 10.00 μmol/L NaAsO2,the levels of tail DNA％ (11.83 ± 1.15,16.85 ± 2.52,24.23 ± 2.75) and olive tail moment (OTM,10.90 ± 1.13,16.19 ± 2.26,23.83 ± 2.79)were significantly increased compared with those of the control group (0.00 μmol/L,2.40 ± 0.51,2.26 ± 0.40,all P ＜ 0.05).After treated with 10.00 μmoFL NaAsO2 for 12 and 24 h,compared with the control group (0 h,3.66 ± 1.02,3.38 ± 1.00),the degrees of tail DNA％ (15.51 ± 1.92,24.18 ± 2.42) and OTM (13.58 ± 2.04,23.14 ± 2.11)were significantly increased (all P ＜ 0.05).②Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the levels of H3K36me3 modifications (60.59 ± 9.75,57.82 ± 11.28,39.45 ± 7.09) were lower at the dosages of 2.50,5.00 and 10.00 μmol/L NaAsO2 (all P ＜ 0.05).Compared with the control group (0 h,100.00 ± 0.00),the levels of H3K36me3 modifications (48.47 ± 9.67,47.75 ± 6.98) were lower after treated with 10.00 μ mol/L NaAsO2 for 12 and 24 h (all P ＜ 0.05).③The levels of H3K36me3 modifications in HaCaT cells exposed to different doses of NaAsO2 were negatively associated with the tail DNA％ and OTM (r =-0.897,-0.903,all P ＜ 0.05).④Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the mRNA levels of MGMT gene were lower at the dosages of 2.50,5.00 and 10.00 pmol/L NaAsO2 (78.20 ± 3.50,61.40 ± 2.60,49.15 ± 4.70,all P ＜ 0.05).⑤There was no observed H3K36me3 enrichmem regularity in the gene encoding ChIP1 and ChIP2 regions of MGMT gene in all doses of NaAsO2 groups (all P ＞ 0.05).Conclusions H3K36me3 may be involved in the regulation of arsenicinduced DNA damage in HaCaT cell.Amenic could inhibit the mRNA transcription of MGMT gene in HaCaT cells,but the transcription of MGMT gene regulate by H3K36me3 is not closely related to DNA damage induced by arsenic.

OBJECTIVE: To assess the effect of one-stage repair of secondary nasolabial deformity and nasal septoplasty for cleft patients on nasal airway resistance (NAR). METHOD: Using active anterior rhinomanometry, NAR was measured in eighteen patients with cleft lip and palate who suffered form one-stage repair of secondary nasolabial deformity and septoplasty at per-and-post operation. RESULT: NAR was (0.664 +/- 0.200) kPa/(s x L) before operation, (0.304 +/- 0.180) kPa/(s x L) six months after operation, and (0.396 +/- 0.250) kPa/(s x L) twelve months after operation respectively. The differences are statistically significant (P < 0.01) between the NAR before and after operation. Subjective impression score of nasal patency was 7.5 +/- 1.5 before-operation, 2.1 +/- 2.0 after-operation for six months, 3.0 +/- 2.4 after-operation for twelve months. There are significant differences in the subjective impression score of nasal patency as well (P < 0.01). CONCLUSION Correction of septal deformities play a very important role in the operation for secondary nasolabial deformity, which can decrease NAR and improve the subjective impression of nasal patency.
Adolescent ; Airway Resistance ; Cleft Lip ; surgery ; Cleft Palate ; surgery ; Female ; Humans ; Male ; Nasal Septum ; abnormalities ; surgery ; Nasopharynx ; surgery ; Postoperative Period ; Rhinoplasty ; Young Adult

Objective To investigate the differences in susceptibility to Lewis lung carcinoma and T lymphocyte subsets in the immune microenvironment between young and elderly mice.Methods Six C57/B6 mice at two months(young)and six mice at twelve months(aged)were injected with Lewis lung carcinoma cells at the dose of 1 × 106 in the left armpit to establish a murine model of lung carcinoma.The weight and tumor growth were monitored.Blood samples for routine blood tests were collected after 24 days.The proportions of CD4+ and CD8+T cells in the spleen were detected by flow cytometry,and the infiltration of CD4+,CD8+ T cells and related effector T cells in the tumor microenvironment were determined in the same way.Results The body weight of tumor bearing mice in the aged group was significantly higher than that in the young group(P ＜0.001);The tumor weight in the aged group(5.084±0.528)g was significantly higher than that in the young group(2.963 ±0.378)g(t =3,349,P =0.012);Routine blood tests showed that the numbers of leukocytes and subsets(except mononuclear)in the aged group were significantly lower than in the young group(P ＜0.05);Flow cytometry found that the effector and memory/effector CD4+T cell ratios in the spleen were significantly higher in the aged group than in the young group(P ＜0.001)and the expression of effector and memory/effector CD8+T cells in the tumor microenvironment was also significantly higher than in the young group(P ＜0.05);Quantitative expression values of IL-6 and IL-10 in the tumor microenvironment were 25090±3820 and 10670± 1793 in the aged group and 6252±864 and 3061±451 in the young group,respectively.Moreover,the expression levels of IL-6 and IL-10(t =3.925,P =0.01;t =3.552,P =0.02)in the tumor microenvironment in the aged group were significantly lower than those in the young group.Conclusions Young mice are more susceptible to Lewis lung carcinoma,probably as a result of differences in inflammation and immunity.