This study was aimed to observe effects of Albizia Julibrissin Flower Total Flavonoids (AJFTF) on the learning and memory abilities and plasma monoamine neurotransmitters 5-HT and NE content of depression model rats . A total of 90 SD rats were randomly divided into the normal group , model group , western medicine group ( Venlafaxine Hydrochloride 0 . 0125 g?kg-1 ) and high , middle and low dosage groups of AJFTF ( equivalent to the crude drug of 10 , 5 , 2 . 5 g?kg-1 ) . Depression model rats were induced by solitary cultiva-tion and chronic unpredictable stress . The learning and memory abilities of model rats were evaluated with the Morris water maze . And the plasma contents of 5-HT and NE were determined by enzyme-linked immunosor-bent assay ( ELISA ) . The results showed that AJFTF ( administered with high , middle and low dosage for 21 days ) significantly shortened the incubation period of the Morris water maze positioning and navigation ( P <0 . 05 or P < 0 . 01 ) , and increased the times of crossing the platform in space exploration ( P < 0 . 05 or P <0 . 01 ) . There was a certain dose-effect relationship . The AJFTF ( high , middle and low dosage ) increased the plasma contents of 5-HT and NE ( P < 0 . 05 ) . It was concluded that AJFTF can improve learning and memory abilities of depression model rats which indicates that AJFTF have potential antidepressant effects . Its action mechanism may be related to increase plasma monoamine neurotransmitter content of 5-HT and NE .

OBJECTIVE To investigate the productive rate of AmpC ?-lactamases and ESBLs produced by the Gram-negative bacilli from the nosocomial infection in our hospital. METHODS AmpC ?-lactamases and ESBLs were detected by the improved cefotaxime and ceftriaxone three-dimensional test. RESULTS The productive rate of AmpC ?-lactamases was 16.00%.Among them,the productive rate of AmpC ?-lactamases was only 8.84%.The Enterobacter cloacae,Serratia marcescens and Enterobacter aerogenes were easy to produce the enzymes(36.00%,31.25% and 28.00%).The productive rate of AmpC ?-lactamases and ESBLs at the same time was 7.16%. CONCLUSIONS The productive rate of the enzymes by the Gram-negative bacilli from the nosocomial infection is rather high.

Background Recently researches indicated that estrogen plays important role in maintaining the normal metabolism of lens. Objective This study was to investigate the changes of estrogen receptor( ER ) α and β expressions in lens upon estrogen level in castrated female rat. Methods Sixty clean adult female Wistar rats were randomized into castrated group,sham operation group,ovariectomy group,ovariectomy with low-dose estradiol eyedropping group,ovariectomy with high-dose estradiol eyedropping group,ovariectomy with low-dose estradiol injecting group and ovariectomy with high-dose estradiol injecting group,and 10 rats for each.The castrated animal models were established by ovariectomy for 5 months.Then 50％,100％ oestradiol benzoate eyedrops were used 4 times per day respectively and 0.2 or 0.4 mg/kg oestradiol benzoate were intramuscularly injected at two-day interval for 6 weeks in corresponding experimental group.Serum estradiol concentration was detected in the rats of various groups at 5 months after ovariectomy and 6 weeks after administration of estradiol benzoate.The animals were sacrificed using the excessive anesthesia method and the lenses were obtained for the assay of ERα and ERβ expressions.The use of the animals complied with the Statement of ARVO. Results No obvious opacification of lenses and the changes of structure and morphology in lens were seen in the rats of various groups under the slit lamp microscope and light microscope during the observing duration after ovariectomy.The significant differences were found in serum estradiol concentrations among the 6 groups ( F=15490.527,P=0.000) or between before and after usage of estradiol benzoate( F=943.236,P =0.001 ).Six weeks after usage of estradiol benzoate,the expressions of ERα and ERβ in the lenses were lower in the castrated group,ovariectomy with high-dose estradiol eyedropping group and ovariectomy with low-dose estradiol injecting group compared with the the sham operative group (P＜0.05),but those in the ovariectomy with low-dose estradiol eyedropping group and ovariectomy with high-dose estradiol injecting group were elevated in comparison with above groups( P＜0.05 ),and expressions of ERα and ERβ in the lenses were similar to the sham operative group ( ERα:28.04±6.80 vs.31.30±7.11 ;ERβ:27.75±7.13 vs.25.38±5.59).Mean A values of ERα and ERβ in the lenses were lower in the castrated group,ovariectomy with high-dose estradiol eyedropping group and ovariectomy with low-dose estradiol injecting group compared with the sham operative group (P＜0.05),but those in the ovariectomy with low-dose estradiol eyedropping group and ovariectomy with high-dose estradiol injecting group were elevated in comparison with above groups ( P＜0.05 ),and mean 4 values of ERα and ERβ in the lenses were similar to the sham operative group (ERα:0.1833 ±0.0087 vs.0.1859 ±0.0067; ERβ:0.1689±0.0059 vs.0.1686±0.0095). Conclusions The expressions of ERα and ERβ in the LECs are associated with the level of serum estradiol.The effects of estrogen on lens were different by different medication way.Low-dose estradiol eyedropping was a more feasible approach to the prevention of cataract.

OBJECTIVETo discuss the protective effect of alkaloids from Piper longum (PLA) in rat dopaminergic neuron injury of 6-OHDA-induced Parkinson's disease and its possible mechanism.

METHODThe rat PD model was established by injecting 6-OHDA into the unilateral striatum with a brain solid positioner. The PD rats were divided into the PLA group (50 mg x kg(-1) x d(-1)), the madorpa group (50 mg x kg(-1) x d(-1)) and the model group, with 15 rats in each group. All of the rats were orally given drugs once a day for 6 weeks. Meanwhile, other 15 rats were randomly selected as the sham operation group, and only injected with normal saline in the unilateral striatum. The behavioral changes were observed with the apomorphine (APO)-induced rotation and rotary rod tests. The number of tyrosine hydroxylase (TH)-positive cells in rat substantia nigra and the density of TH-positive fibers in striatum were detected by tyrosine hydroxylase immunohistochemistry. The content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), catalase (CAT), malondialdehyde (MDA), nitric oxide (NO) and nitric oxide synthase (NOS) in rat substantia nigra and striatum were measured by the spectrophotometric method.

RESULTAfter being induced by APO, PD rats showed obvious rotation behaviors, with decreased time stay on rotary rod and significant reduction in the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. The activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity significantly decreased, whereas the activities of NOS and the content of MDA, NO significantly increased. PLA could significantly improve the behavioral abnormality of PD rats and increase the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. It could up-regulate the activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity, and decrease the content of NOS and the content of MDA, NO.

CONCLUSIONAlkaloids from P. longum shows the protective effect in substantia nigra cells of 6-OHDA-induced PD model rats. Its mechanism may be related with their antioxidant activity.

Administration, Oral ; Alkaloids ; administration & dosage ; pharmacology ; Animals ; Apomorphine ; pharmacology ; Catalase ; metabolism ; Dopamine Agonists ; pharmacology ; Dopaminergic Neurons ; drug effects ; metabolism ; pathology ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Motor Activity ; drug effects ; Neostriatum ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; physiopathology ; prevention & control ; Piper ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism ; Tyrosine 3-Monooxygenase ; metabolism

Type 1 diabetes mellitus (T1D) is an immune-mediated disease. The autoreactive T cells in T1D patients attack and destroy their own pancreatic cells. In order to systematically investigate the potential autoreactive T cell receptors (TCRs), we used a high-throughput immune repertoire sequencing technique to profile the spectrum of TCRs in individual T1D patients and controls. We sequenced the T cell repertoire of nine T1D patients, four type 2 diabetes (T2D) patients and six nondiabetic controls. The diversity of the T cell repertoire in T1D patients was significantly decreased in comparison with T2D patients (P = 7.0E
08 for CD4+ T cells, P = 1.4E
04 for CD8+ T cells) and nondiabetic controls (P = 2.7E
09 for CD4+ T cells, P = 7.6E
06 for CD8+ T cells). Moreover, T1D patients had significantly more highly-expanded T cell clones than T2D patients (P = 5.2E
06 for CD4+ T cells, P = 1.9E
07 for CD8+ T cells) and nondiabetic controls (P =1.7E
07 for CD4+ T cells, P= 3.3E
03 for CD8+ T cells). Furthermore, we identified a group of highly-expanded T cell receptor clones that are shared by more than two T1D patients. Although further validation in larger cohorts is needed, our data suggest that T cell receptor diversity measurements may become a valuable tool in investigating diabetes, such as using the diversity as an index to distinguish different types of diabetes.

ABSTRACT:OBJECTIVE To investigate the effects of Whenshen Zhixie Fang administrated through bellybutton and the in-fluence on 5-HT and NO of spleen and kidney yang deficiency type IBS-D patients.METHODS By using computer,parallel group and non-blinded trials,patients were randomly divided into two groups.58 cases in the treatment group were given Wen-shen Zhixie Fang administrated through bellybutton together with orally taken Montmorillonite powder.The control group was given orally taken Montmorillonite powder only.The treating effects were assessed after ten days.RESULTS Total effective rate of the treatment group was 82.76%,which was better than that of the control group (66.10%)(P <0.05).Abdominal pain and discomfort were significantly relieved in the treatment group,and the shape of the stool and frequency were also changed for better(P <0.01),which were better than those of the control group except the stool.And 5-HT was significantly decreased and NO was increased,which were better than those of the control group(P < 0.01).CONCLUSION Wenshen Zhixie Fang administered through Bellybutton can improve the clinical symptoms of IBS-D and its mechanism may be related to the regulation of 5-HT and NO levels and reduction of intestinal sensitivity.

Objective To study the inhibitory effect of Tilianin on non -small cell lung cancer cell lines (A549) and the associated mechanisms.Methods The control group and 10,20,40,80 and 160 μmol· L-1 experimental group were treated with 0,10,20,40,80 and 160 μ mol· L-1Tilianin for 24 h,cell proliferation and toxicity test kit (CCK8) was used to observe the proliferation of A549 cells.The apoptosis of A549 cells in each group was detected by flow cytometry.In the hypoxia model group,A549 cells were cultured in hypoxic incubator for 4 h.In the hypoxia experimental group of 10,20 and 40 μmol · L-1Tilianin,A549 cells were pretreated with 10,20 and 40 μmol · L-1Tilianin for 4 h,and then cultured in hypoxic incubator for 4 h.Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the gene expressions of vascular endothelial growth factor(VEGF) and HIF-1α (HIF-1α).The protein expressions of VEGF-A,HIF-1α and p-Akt were detected by Western blot.Results Compared with control group,Tilianin significantly inhibited the proliferation of A549 in a dose -dependent manner.The inhibitory rates of proliferation of 10,20,40,80 and 160 μmol · L-1 experimental groups were (5.83 ±1.67)％,(6.77 ±0.87)％,(12.26 ±0.23)％,(22.97 ±0.50)％ and (46.24±1.44)％,the difference was statistically significant (P ＜0.05).The apoptosis rates of 10,20,40,80 and 160 μmol · L-1Tilianin were (6.80 ±0.62)％,(14.70 ±1.36)％,(24.76 ±4.37)％,(39.26±6.42)％ and (62.31 ±1.79)％,respectively,compared with the control group,the differences were statistically significant (P ＜ 0.05).In the hypoxia model group,the gene expression of HIF-1α was 4.30 ± 0.26,VEGF expression was 6.02 ± 0.53,compared with the control group,the differences was statistically significant (P ＜0.05).In 10,20,40 μmol · L-1Tilianin hypoxia experiment group,the gene expression of VEGF were 4.73 ±0.20,2.31 ±0.09 and 1.47 ±0.16,respectively.The expression of HIF-1α were 3.01 ± 0.11,1.81 ± 0.13 and 1.03 ± 0.16.Compared with hypoxia model group,the differences was statistically significant (P ＜ 0.05).The expression of p-AKT protein in hypoxic model group was (106.47 ± 2.08)％,the expression of HIF-1αwas (204.31 ± 8.35)％,the expression of VEGF-A was 212.30 ±4.80.The expression of p-AKT protein in 10,20,40 μmol · L-1 hypoxic experimental group were (87.51 ±2.72)％,(75.18 ± 1.67)％ and (32.40 ± 0.86)％,the protein expression levels of HIF-1α were (182.54 ± 6.42),(90.95 ± 2.76)％,(15.03 ± 4.21)％,the protein expression of VEGF-A were (156.38 ± 5.12) ％,(79.62 ± 2.84) ％ and (13.72 ± 4.64) ％,with significant difference (P ＜ 0.05).Conclusion Tilianin has the effect of inhibiting the proliferation and inducing apoptosis of A549 in vitro through AKT / HIF-1α signaling pathway.

OBJECTIVETo investigate the effect of air pollution on respiratory health in school-aged children in the main urban area of Chongqing, China.

METHODSThe main urban area of Chongqing was divided into polluted area and clean area according to the air pollution data shown on the Environmental Protection Agency Website of Chongqing between 2010 and 2015. A cluster sampling method was used to select 695 third- or fourth-grade children from 2 primary schools in the clean or polluted area as study subjects, with 313 children from the clean area and 382 children from the polluted area. Pulmonary function was examined for all children and a standard American epidemiological questionnaire (ATS-DLD-78-C) was used to investigate the prevalence of respiratory diseases and symptoms.

RESULTSCompared with the clean area, the polluted area had significantly higher concentrations of inhalable particles (PM), fine particulate matter (PM), and nitric oxide (NO) (P<0.05). The multivariate logistic regression analysis was performed after adjustment for confounding factors, and the results showed that compared with those in the clean area, the children in the polluted area had significantly higher risks of cough (OR=1.644), cough during cold (OR=1.596), expectoration during cold (OR=2.196), persistent expectoration (OR=1.802), and wheezing (OR=2.415). The boys and girls in the clean area had significantly higher forced vital capacity and forced expiratory volume in one second than those in the polluted area (P<0.05).

CONCLUSIONSAir pollution in the main urban area of Chongqing is associated with the increased prevalence of respiratory symptoms in school-aged children and has certain effect on children's pulmonary function.

Air Pollution ; adverse effects ; Child ; Female ; Forced Expiratory Volume ; Humans ; Logistic Models ; Male ; Respiratory Tract Diseases ; etiology ; Vital Capacity

BACKGROUNDDengue is currently a significant global health problem but no vaccines are available against the four dengue serotypes virus infections. The development of safe and effective vaccines has been hampered by the requirement of conferring complete protection against all four dengue serotypes and the lack of a convenient animal model. Virus-like particles (VLPs) have emerged as a promising subunit vaccine candidate. One strategy of vaccine development is to produce a tetravalent dengue subunit vaccine by mixing recombinant VLPs, corresponding to all four dengue virus serotypes. Towards this end, this study aimed to establish a Pichia pastoris (P. pastoris) expression system for production of dengue virus type 1 (DENV-1) VLPs and evaluate the humoral and cellular immune response of this particle in mice.

METHODSA recombinant yeast P. pastoris clone containing prM and E genes of DENV-1 was constructed and DENV-1 VLPs expressed by this clone were analyzed by sucrose density gradient centrifugation, Western blotting, and transmission electron microscope. Groups of mice were immunized by these particles plus adjuvant formulations, then mice were tested by ELISA and neutralization assay for humoral immune response, and by lymphocyte proliferation and cytokine production assays for a cellular immune response.

RESULTSOur data demonstrated that recombinant DENV-1 VLPs consisting of prM and E protein were successfully expressed in the yeast P. pastoris. Sera of VLPs immunized mice were shown to contain a high-titer of antibodies and the neutralization assay suggested that those antibodies neutralized virus infection in vitro. Data from the T lymphocyte proliferation assay showed proliferation of T cell, and ELISA found elevated secretion levels of interferon IFN-γ and IL-4.

CONCLUSIONSP. pastoris-expressed DENV-1 VLPs can induce virus neutralizing antibodies and T cell responses in immunized mice. Using P. pastoris to produce VLPs offers a promising and economic strategy for dengue virus vaccine development.

Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Dengue Virus ; genetics ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Male ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; metabolism ; T-Lymphocytes ; immunology

BACKGROUNDThe potential application of retinoic acid receptor activators, such as all trans-retinoic acid (ATRA), for treating various cancers have been studied both pre-clinically and clinically. Whether ATRA has an anticancer effect on human esophageal squamous cancer cell (ESCC) is still unknown. We have explored the anticancer effect of ATRA in ESCC, and in this study, the effects of ATRA on levels and patterns of expression of the vascular endothelial growth factor (VEGF) signal transduction pathway in transplantable tumor growth of the human ESCC cell line, EC9706, in nude mice.

METHODSThe animal model of the ESCC xenograft was made by subcutaneous implantation of tumor cells into nude mice. Reverse transcription-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical assays were used to detect the expression of the VEGF signal transduction pathway in ESCC xenograft tissues.

RESULTSCompared to the control group, the tumor inhibition rates in the low dose ATRA, high dose ATRA, and 5-FU groups were 83.21%, 88.32%, 91.02%, respectively. The protein and mRNA levels of VEGF were down-regulated after being treated with ATRA and 5-FU compared to the control group (P < 0.05). The study also revealed that ATRA specifically down-regulated VEGF and the component of the VEGF signal transduction pathway of CD31, CD34, and CD105 (component of the TGF-β receptor) in ESCC xenograft tissues (P < 0.05).

CONCLUSIONSATRA can significantly inhibit tumor growth and has anticancer effects on transplantable tumor growth of human ESCC cell line EC9706 in nude mice. These findings indicate that ATRA specifically down regulated VEGF and the components of VEGF signal transduction, which may be an important mechanism responsible for the neoangiogenesis inhibition of ESCC cells.

Animals ; Blotting, Western ; Carcinoma, Squamous Cell ; drug therapy ; metabolism ; Cell Line, Tumor ; Esophageal Neoplasms ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Neovascularization, Pathologic ; drug therapy ; metabolism ; Real-Time Polymerase Chain Reaction ; Tretinoin ; therapeutic use ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays