Objective To investigate the influence of the eye acupuncture therapy in the expressions of brain derived neurophic factor(BDNF)and tyrosine kinase receptor B(TrkB)in penumbra brain tissue of ischemic area in the rats with acute cerebral ischemia reperfusion inj ury,and to explore the related mechanism of protective effect on brain of eye acupunture.Methods 62 SD rats were randomly divided into blank control group (n=11),sham operation group (n=11)and model copy group (n=40).The rats in copy model group were used to establish models with suture method,and the successful model rats (n=32)were randomly divided into model group(n=16)and eye acupuncture group(n=16).The rats in eye acupuncture group were treated with eye acupuncture intervention;the acupoints were chosen according to human acupoint selection method, the liver, the upper-jiao, the down-jiao district,and renal were selected for acupuncture intervention. After reperfusion for 2 h,the acupuncture was performed once every 8 h,lasted for 10 times;30 min after the last intervention,the rats were sacrificed;the regional ischemia penumbra around the parts of the brain tissue was obtained and the expressions of BDNF and TrkB mRNA and protein in brain tissue of the rats were detected with RT-PCR and Western blotting method. Results Compared with blank control group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in sham operation group had no statistical significance(P>0.05);compared with sham operation group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in model group were significantly up-regulated(P<0.01);compared with model group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in eye acupuncture group were significantly up-regulated(P<0.05 or P<0.01).Conclusion The expressions of BDNF and TrkB are increased after cerebral ischemia and reperfusion injury.Eye acupuncture can protect the brain of MCAO/R rats,which may be related to the up-regulation of the expressions of BDNF and TrkB in the corresponding penumbra.

Objective to observe the influence of Candida albicans infection on t cell subsets in mice with spleen deficiency,so as to explore the immune mechanism. Methods totally 150 healthy SPF mice were randomly divided into four groups:control group(N1 group,n = 30);Candida albicans infection group(N2,n = 40),spleen deficiency model group(M1,n = 40),spleen deficiency model combined with Candida albicans in-fection group(M2,n = 40). N2 and M1 mice were infected by Candida albicans at a concentration of 2×108 CFU/mL. ten mice were randomly se-lected from each group at 7,14,and 21 days after infection respectively,and the index was detected. Flow cytometry was used to detect the percent-age of CD4+/CD8+t cells in intestinal mucosa of mice,and the expression level of IL-4 and IFN-γ mRNA was detected by Rt-PCR assay. the levels of IFN-γ and IL-4 were detected by Western blot assay. Results Compared with N1 group,the proportion of CD4+t cells in the lamina of the natu-ral layer of the small intestine was decreased(P < 0.01),the proportion of CD8+t cells was increased(P < 0.01),the ratio of the two was significant-ly decreased(P < 0.01),and the expression levels of IL-4,mRNA IFN-γ and protein in the small intestine tissues were increased in other groups (P < 0.01). In addition,the expression level of IFN-γ was significantly increased in M2 group,while the expression of IL-4 was significantly in-creased in N2 group. Conclusion the susceptibility of Candida albicans infection was increased in spleen deficiency mice,which may be closely related to the regulation of th1/th2 balance.

Objective To investigate the effects of Erlong Zuoci Pills on AQP4 expression in cochlear tissue of mice with elderly kidney deficiency deafness;To discuss the action mechanism. Methods Intraperitoneal injection of hydrocortisone method was used to duplicate mice models with kidney deficiency except the normal control group. After the models were established, mice were divided into model group and TCM group, 16 mice in each group. TCM group was gavaged by Erlong Zuoci Pills, model group and normal control group were gavaged by normal saline for 22 d. Cochlear stretched preparation technology was used to observe morphological changes in cochlear inner and outer hair cells, and supporting cells. Immunohistochemistry and Western bolt were used to detect protein expression of AQP4. Results Compared with normal control group, mice in model group missed inner and outer hair cells and supporting cells of the cochlea. Compared with model group, arrangement of cochlear inner and outer hair cells and supporting cells was neat and boundary was clear in TCM group. Compared with model group, protein expression of AQP4 in cochlear tissues in TCM group increased (P<0.01). There was no difference between TCM group and normal control group. Conclusion Erlong Zuoci Pills have significant therapeutic effect for elderly kidney deficiency deafness, and the treatment is related to the upregulation of protein expression of AQP4 in cochlear tissues.

Objective To explore the effect of tert?butylhydroquinone(tBHQ)on ultraviolet B(UVB)?induced oxidative damages in human im?mortalized keratinocytes(HaCaT),and discuss its mechanism. Methods The cultured HaCaT cells were randomly divided into 4 groups:control group(G1),ultraviolet irradiation group(G2),25μmol/L tBHQ pretreatment before ultraviolet irradiation group(G3),and 50μmol/L tBHQ pre?treatment before ultraviolet irradiation group(G4). The content of reactive oxygen species was detected by DCFH?DA method,and the cell prolifera?tion was evaluated by MTT. Western blot was used to measure the protein expression of nuclear factor E2?related factor 2(Nrf2)in both nuclear fac?tions and whole?cell of HaCaT. The mRNA expressions of CAT and SRX were determined by real?time RT?PCR. Results The content of reactive oxygen species in HaCaT cells was increased,and the cell proliferation rate was decreased significantly after ultraviolet irradiation. The pretreatment of 25 and 50μmol/L tBHQ can inhibit the UVB?induced oxidative damage in a dose?dependent manner in HaCaT cells. Compared with G2 group, tBHQ pretreatment could dose?dependently increase the level of Nrf2 protein in nuclear factions and whole?cell of HaCaT,and also the mRNA ex?pressions of CAT and SRX. Conclusion UVB irradiation can induce oxidative stress damages of HaCaT cells. tBHQ may inhibit the UVB?induced oxidative damages through enhancing Nrf2 expressions and nuclear translocation,then activating the transcription of the downstream antioxidant en?zymes CAT and SRX.

Objective To observe the effects of Jianpi Yangxue Qufeng Formula (JPYXQF) on the AQP3 in mice with chronic eczema, and explore mechanism of action. Methods Fifty healthy male mice were randomly divided into 5 groups, namely normal group, model group, positive medicine group and JPYXQF high and low dose groups. Low-dose DNCB and Sennae Fominm were used to establish mice models of chronic eczema with spleen deficiency. JPYXQF groups were treated by JPYXQF for gavage, while the positive medicine group was treated by levocetirizine hydrochloride for gavage. The expression of AQP3 in mice skin tissue was detected by immunohistochemical method. At the same time, the pathological changes of skin were observed. Results The pathology of mice skin lesion showed that JPYXQF has certain recovery effects on the inflammation injury of skin lesion. Compared with the normal group, expression of AQP3 over expressed in model group. Compared with the model group, the expression of AQP3 in all treatment groups significantly decreased, and the staining intensity decreased. In the model group, the average optical density of AQP3 was significantly higher than that in the normal group (P<0.01). Compared with the model group, the treatment groups can reduce the expression of AQP3 in mice skin tissues (P<0.05). Conclusion JPYXQF can reduce the over expression of AQP3 in skin lesion, which is probably its mechanism for the treatment of chronic eczema.

Objective To investigate the pathogen culturing of the catheter related infection(CRI),cathe-ter related bloodstram infection(CRB)and risk factors after central venous catheter(CVC)of cardiovascular surgery in order to provide the beneficial reference.Methods From Jan 2005 to Dec 2005,a total of 300 cases central ve-nous cathers were determined,and the cusp of the catheters was determined by bacteria cultivation,and blood bacte-ria cultivation.Results The infection happened in 35 of 300 patients with inserted central venous catheter.The cusps of CRI rate was 11.7%.CRB rate was 1.7%.54.3%pathogens were gram-positive cocci,34.3% were gram-negative bacilli,11.4% were fungi.The most common strain were Staphylococcus epidermis,Staphylococcus aureus,Klebsiella pneumoniae,Pseudomonas aeruginose,and Candiadia albicans.The infection rate increased obviously when the dwelling time>6 d.Conclusion CRI and CRB are the most severe complication of CVC,and it is important to cut down the death with the early diagnosis and applying antibiotics rationally.

Objective To investigate the effect of the eye-acupuncture therapy on serum TNF-α levels in rats with cerebral ischemia-reperfusion injury and the related mechanism. Methods Healthy SD rats were randomly divided into four groups: normal group, sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group according to body weight. Sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group were divided into the 3h group, the 24h group and the 72h group, a total of 10 groups (n=8). To establish the rat model of cerebral ischemia-reperfusion by suture method in ischemia-reperfusion model group and eye-acupuncture group. Eye-acupuncture was separately started immediately after reperfusion and at 30 min before sampling in the 3h eye-acupuncture group, besides, eye-acupuncture was separately taken every 12h in the 24h eye-acupuncture group and in the 72h eye-acupuncture group. The rats in normal group were not treated, those in sham-operated control group were inserted the fishing thread 1cm, and the others were identical with those in ischemia-reperfusion model group. At 3h, 24h, 72h after reperfusion, the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks in ischemia-reperfusion model group and eye-acupuncture group. The method of ELISA was taken to detect the change of serum TNF-α levels in rats after the eye-acupuncture therapy. Results Compared with the ischemia-reperfusion model group, the neurologic impairment score of the eye-acupuncture group decreased obviously; The levels of serum TNF-αin ischemia-reperfusion model group at 3 h, 24 h, 72 h respectively were(76.803±18.325)pg/ml、(85.511±13.334)pg/ml、(86.831±9.232)pg/ml. The level of serum TNF-α in normal group was(24.304±6.511)pg/ml. The level of serum TNF-α in Sham-operated control group at 3 h, 24 h, 72 h respectively were(24.928±3.792)pg/ml,(27.533±5.362)pg/ml,(29.366±5.874)pg/ml. Ischemia-reperfusion model group compare with normal group and sham-operated control group, the difference were significant(P＜0.01). The levels of serum TNF-α in rats after the eye-acupuncture therapy at 3 h, 24 h, 72 h respectively were(40.185±3.335)pg/ml, (48.523±7.687)pg/ml, (51.611± 6.403)pg/ml. Compared with ischemia-reperfusion model group, the levels of serum TNF-α were strongly reduced(P＜0.01). Conclusion The eye-acupuncture therapy could play a role in improving cerebral ischemia-reperfusion injury evidently and the mechanism was related to its reducing serum TNF-α levels.

2-Keto-L-gulonic acid (2-KLG), the precurcor of L-ascorbic acid synthesis, was prepared directly from D-glucose by tandem fermentation. In the first step fermentation Erwinia sp. SCB 247 translated D-glucose to 2,5-diketo-D-gluconate (2,5-DKG), which accumenlated 180 mg 2,5-DKG per ml in the broth. In the second step fermentation Corynebacterium sp. SCB 3058 reduced 2,5-DKG to 2-KLG, which accumulated 35 mg 2-KLG per ml in the broth. This reductive fermentation was obtained under aerobic conditions by adding a hydrogen donor such as glucose.The average yield of five batches fermentation was 56.3 mol%, from D-glucose to 2-KLG in 10 L fermentor.

Objective To explore the effect and potential mechanism of Huayu Mingmu Recipe(HMR)-containing serum on high glucose-induced angiogenesis of human retinal microvascular endothelial cells(HRMECs).Methods CCK-8 method is used to screen the optimal sugar concentrations,as well as volume fraction of drug-containing serum.A model of high glucose-induced HRMECs dysfunction was established.The HRMECs were divided into 6 groups such as normal control group which was treated with normal ECM culture medium(containing 5.5 mmol·L-1 glucose)and 10%blank serum,the mannitol control group and the high glucose model were given 19.5 mmol·L-1 mannitol and 19.5 mmol·L-1 D-glucose on the basis of the treatment of normal control group,respectively.The low-,medium-,and high-dose groups of HMR were given 10%low-,medium-,and high-dose medicated serum(without blank serum)on the basis of the model group.The colony experiment was used to detect the number of cell colonies.Transwell experiment was used to test the number of cell migration,and tube formation experiment was used to determine the forming of tubes.Immunofluorescence,Western Blot and RT-PCR were used to detect levels of protein and mRNA expression of FactorⅧ,platelet endothelial cell adhesion molecule-1(CD31),cell differentiation factor(CD34),vascular endothelial growth factor A(VEGFA),vascular endothelial growth factor receptor 2(VEGFR2).Results Compared with the normal control group,the model group could promote colony formation of HRMECs(P＜0.01),cell migration(P＜0.01),and lumen formation(P＜0.01).The levels of protein and mRNA expressions of Factor VIII,CD31,CD34,VEGFA,VEGFR2 were significantly increased(P＜0.01)in model group.Compared with the model group,low-,medium-and high-dose HMR-containing serum groups could inhibit colony formation of HRMECs(P＜0.05,P＜0.01),cell migration(P＜0.01),and lumen formation(P＜0.05,P＜0.01).The levels of protein and mRNA expressions of Factor VIII,CD31,CD34,VEGFA,VEGFR2 were significantly reduced(P＜0.05,P＜0.01)in HMR-containing serum groups.There was no statistically significant difference in results of various tests between the normal control group and the mannitol control group(P＞0.05).Conclusion HMR-containing serum can inhibit the proliferation,migration,and tube formation of HRMECs induced by high glucose,and then prevent or reduce angiogenesis.The mechanism may be related to the regulation of VEGFA/VEGFR2 signaling pathway.

Objective:To investigate the effect and mechanism of Grifola frondosa extract on inflammatory response of colon tissue in rats with ulcerative colitis(UC)by regulating interleukin-6(IL-6)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods:Forty SD rats were randomly divided into blank control group,UC model group,Grifola frondosa treatment group,western medicine treatment group and combined treatment group,with 8 rats in each group.After UC rats were established by free drinking 3%DSS for 7 days,the treatment group were given Grifola frondosa extract 10 mg/(kg·d),sulfasalazine 0.3 g/(kg·d),and the same amount of two drugs,for 14 consecutive days.During the experiment,general state of rats were observed,and the disease activity index(DAI)score was calculated;pathological changes of rats colon tissue were observed by HE staining;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in rats colon tissue were detected by Western blot;content of IL-6 in rats serum was detected by ELISA;protein contents and expressions of IL-6R and MPO in rats colon tissue were determined by immunohistochemistry.Results:Compared with blank control group,general state of rats in UC model group was poor,DAI score was increased,obvious tissue mucosal defects and inflammatory cell infiltration were observed by HE staining;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in rats colon tissue and contents of IL-6R and MPO were significantly increased(P<0.01);content of IL-6 in rats serum was significantly increased(P<0.01),the difference was statistically significant.Compared with UC model group,general condition of rats in each treatment group was improved,DAI score was decreased,HE staining showed that mucosal defects were improved to varying degrees,and occasionally inflammatory cell infiltration was observed;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in colon tissue were significantly decreased(P<0.01),contents of IL-6R and MPO in colon tissue and content of IL-6 in serum were significantly decreased(P<0.01 or P<0.05),the differences were statistically significant.Conclusion:Grifola frondosa extract can reduce the inflammatory response in colon tissue of UC rats by regulating expressions of IL-6/JAK2/STAT3 signaling pathway related factors.