AIM:To explore the effect of long-term (6 months) endogenous testosterone deprivation by orchidectomy on the function of voltage-dependent potassium channels of vascular smooth muscle cells in rats. METHODS:Wistar rats were raised for 6 months after castration. Isometric tension measurement of aortic rings,whole-cell patch-clamp technique and Western blotting analysis were employed to examine the functional and posttranscriptional alterations of voltage-dependent potassium channels. RESULTS:Voltage-dependent potassium channel blocker,4-aminopyridine,significantly decreased the constriction of aortic artery rings from male rats after 6-month castration. In castrated rats the amplitude of voltage-dependent potassium currents of aortic artery smooth muscle cells was significantly decreased compared with that in control rats. Meanwhile,the expression of Kv 1.5 channel protein,which plays an essential role in mediating vasomotor function,was also reduced. The functional and molecular alterations of voltage-dependent potassium channels were both restored when the rats were concomitant applied with physiological level of testosterone after castration. CONCLUSION:Long-term deprivation of endogenous testosterone in rats significantly attenuates the function of voltage-dependent potassium channels,and the decreases in expression of Kv1.5 channel protein accounts for this alteration. Long-term application of physiological concentration of testosterone,which recovered the impaired function of voltage-dependent channels,may be beneficial for male gender with hypotestosteronaemia.

OBJECTIVETo observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.

RESULTSRP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).

CONCLUSIONSHJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Animals ; Berberine ; Berberine Alkaloids ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavanones ; Flavonoids ; Glucose ; metabolism ; Iridoids ; Models, Animal ; Neurons ; Oxygen ; metabolism ; Rats ; Reperfusion Injury ; drug therapy

OBJECTIVETo study the effect of Modified Dachengqi Decoction (MDD) as whole course therapy on mediators of inflammation in severe acute pancreatitis (SAP) model rats, and to compare interventional advantages over intestinal mucosal barrier (IMB) of SAP rats between whole course therapy of MDD and early stage therapy of MDD.

METHODSTotally 190 SD rats were divided into five groups according to random digit table, i.e., the sham-operation group, the model group, the octreotide (OT) group, the early stage MDD treatment group, the whole course MDD treatment group, 38 in each group. SAP models were established with retrograde injection of 5% sodium taurocholate into the pancreaticobiliary duct. Three hours after modeling normal saline (NS) was administered to rats in the sham-operation group and the model group by gastrogavage, once per 12 h.1.35 µg/100 g OT was subcutaneously injected to rats in the OT group, once every 8 h. 0.4 mL/100 g MDD was administered to rats in the early stage MDD treatment group, and 6 h later changed to NS (once per 12 h).0.4 mL/100 g MDD was administered to rats in the whole course MDD treatment group, once every 12 h. The accumulative survival rate and morphological manifestations of pancreas and small intestine were observed under microscope 48 h after modeling. Pathologic scores of the pancreas and small intestine were conducted at 4, 6, 24, and 48 h after modeling. Contents of serum amylase (AMY), alanine transaminase (ALT), and TNF-α were also detected. The expression of high mobility group box protein 1 (HMGB1) in the small intestine tissue was also detected by Western blot. The positive rate of bacterial translocation in mesenteric lymph nodes (MLNs) was observed within 48 h. Correlations between serum TNF-α or HMGB1 in small intestinal tissue and pathological scores of the pancreas or the small intestine were analyzed.

RESULTSThe accumulative survival rate was 100. 0% in the sham-operation group, 79. 2% in the whole course MDD treatment group, 70. 8% in the OT group, 45. 8% in the early stage MDD treatment group, and 37.5% in the model group. At 6 h after modeling, pathological scores decreased more in the whole course MDD treatment group, the early stage MDD treatment group, the OT group than in the model group (P < 0.05). At 24 and 48 h after modeling, pathological scores of the pancreas and the small intestine decreased more in the whole course MDD treatment group and the OT group than in the early stage MDD treatment group (P <0. 05). At 6, 24, and 48 h after modeling, serum contents of AMY and ALT both decreased more in the whole course MDD treatment group, the early stage MDD treatment group, the OT group than in the model group (P < 0.05). At 48 h after modeling serum contents of AMY and ALT both decreased more in the whole course MDD treatment group and the OT group than in the early stage MDD treatment group (P < 0.05). At 6 h after modeling serum TNF-α levels decreased more in the whole course MDD treatment group, the early stage MDD treatment group, the OT group than in the model group (P < 0.05). At 6, 24, and 48 h after modeling the level of HMGB1 in the small intestinal tissue decreased more in the whole course MDD treatment group, the early stage MDD treatment group, the OT group than in the model group (P < 0.05). Of them, HMGB1 levels at 24 and 48 h were lower in the whole course MDD treatment group and the OT group than in the early stage MDD treatment group (P < 0.05). The number of MLNs bacterial translocation at 48 h after modeling was lower in the whole course MDD treatment group and the OT group than in the early stage MDD treatment group and the model group (P < 0.05). Serum TNF-α contents within 6 h were positively correlated with pathological scores of pancreas (r = 0.579, P < 0.01). ROC curve showed that serum TNF-α contents could predict the severity of SAP (ROC = 0.990, 95% Cl: 0.971 to 1.000). HMGB1 in the small intestine was positively correlated with pathological scores of the small intestine (r = 0.620, P < 0.01).

CONCLUSIONSEarly stage use of MDD could effectively reduce the release of TNF-α, while whole course use of MDD could effectively inhibit the expression of HMGB1. The latter could preferably attenuate injuries of the pancreas and the small intestine, lower MLNs bacterial translocation, and elevate the survival rate.

Animals ; Bacterial Translocation ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; HMGB1 Protein ; Intestinal Mucosa ; drug effects ; Octreotide ; Pancreas ; Pancreatitis ; drug therapy ; Plant Extracts ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Taurocholic Acid ; Tumor Necrosis Factor-alpha

Objective To observe the expression of Bcl-2,Bax and proliferating cell nuclear antigen(PCNA) in liver of rats with hepatic fibrosis and the effects of transforming growth factor (TGF)-?1 vaccine on them.Methods Thirty healthy male Sprague-Dawley rats were assigned into 3 groups,named healthy control group(n=10),hepatic fibrosis group(n=10) and TGF-?1 vaccine treated group(n=10).The animal model with hepatic fibrosis was established by injecting solution dimethylnitrosamine (DMN) into abdominal cavity with concentration as 0.5% and dose as 0.2 mL/ 100 g.In TGF-?1 vaccine treated group,every rat was not only injected with DMN but also 150?g TGF-?1 vaccine protein.On the 42nd day,all rats were sacrificed.Then the blood and the liver tis- sues were collected.The expression levels of Bcl-2,Bax and PCNA in liver tissues were detected by S -P immunohistochemistry and observed by routine pathological evaluation.Alanine aminotransferase (ALT),aspartate aminotransferase(AST) and albumin(Alb) were determined by auto biochemical analytical tool.Serum levels of hyaluronic acid (HA),laminin(LN) were detected by radioimmunoas- say (RIA).Results The expression of Bax,which promoted apoptosis,directly correlated with pathological grade in liver of rats,while the expression of Bcl-2 and Bcl-2/Bax,which protected a gainst apoptosis,inversely correlated with pathological grade in liver of rats.The expression levels of TGF-?1 and Bax in healthy control group were significantly lower than those of fibrosis group,how ever,the expression levels of Bcl-2 were comparable between these two groups.As compared with fi- brosis group,the expression of TGF-?1 was significantly lower while the expression of Bcl-2 was sig nificantly higher in TGF-?1 vaccine treated group.However,the expression of Bax was comparable between these two groups.The expression level of PCNA of fibrosis group was significantly higher than that of healthy control group but dramatically lower than that of TGF-?1 vaccine treated group (Both P

OBJECTIVETo study the impact of total flavones from Artemisia anomala (TFAS) on activation of macrophages, cell oxidative stress, auto-nitration of CuZn-SOD, platelet aggregation and isolated vascular tension.

METHODLPS and IFN-gamma induced activation of macrophages and oxidative stress in rats; H2O2 and nitrite induced auto-nitration of CuZn-SOD; ADP, AA and collagen induced platelet aggregation in vitro in mice; PE stimulates isolated vascular tension; nitrite content of macrophages was measured by Griess assay; MTT assay and FRAP assay was applied for cell viability and total cell antioxidant capacity; auto-nitration of CuZn-SOD was measured by Western blot and colorimetric methods; platelet aggregation was detected by turbidimetry; and aorta ring relaxation was recorded by isolated vascular function experience devices for rats.

RESULTTFAS demonstrated dose dependence (25, 50, 100, 200 mg x L(-1)) on inhibiting induced macrophages NO production from generating, while increasing cell viability and total anti-oxidant capacity. Auto-nitration of CuZn-SOD was suppressed by TFAS in dose dependence (0.5, 5, 50 mg x L(-1)). TFAS showed an inhibitory effect on collagen-induced platelet aggregation at 50 mg x L(-1) and an endothelium-dependent relaxation effect on PE-induced vasoconstriction at 1 g x L(-1).

CONCLUSIONTFAS shows effect on anti-inflammation, anti-oxidation, anti-nitration, anti-platelet aggregation and vasodilatation in experiment in vitro, which may inhibit vascular inflammatory by regulating multiple target points. It is among material bases for promoting blood circulation and removing blood stasis.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Aorta ; drug effects ; immunology ; physiology ; Artemisia ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Flavones ; administration & dosage ; Humans ; Macrophages ; drug effects ; immunology ; Mice ; Oxidative Stress ; drug effects ; Rats ; Vasodilation ; drug effects

The modern studies indicate that there is a close relationship between mast cells and the acupuncture effect, and acupuncture can activate mast cells to induce a series of vascular reaction and immunological effect. The authors hold that acupuncture is a kind of nociceptive stimulus, which can cause inflammatory reaction in the sites of acupuncture, and then further activate the nerve-endocrine-immune network to cause the cascade amplification of the acupuncture effect. The inflammatory reaction induced by acupuncture is one of the initial factors of acupuncture effect.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Immunity ; Mast Cells ; immunology ; Nerve Fibers ; immunology ; Neuroimmunomodulation ; Skin ; immunology

Objective To study sleep characteristics in patients with temporal lobe epilepsy (TLE) through polysomnography (PSG). Methods Twenty-five TLE patients (TLE group) and eighteen healthy volunteer subjects (control group) were recruited to our study. Patients of two groups were evaluated by whole-night PSG, including total time in bed (TIB), total sleep time (TST), sleep efficiency (SE), sleep latency (SL), rapid eye movement latency (REML), wake after sleep onset (WASO), the percentages of non-REM (NREM) 1, 2 and 3 stages and the percentages of rapid eye movement (REM) occupied TST (N1％, N2％, N3％and REM％), the apnea-hypopnea index (AHI), hypopnea index, mean oxygen saturation (SpO2) and nadir SpO2, periodic leg movements (PLMs) index and PLMs index of REM sleep, sleep stage shifts (SSS) and sleep stage shifts per hour (SSS/h), NREM1, NREM2, NREM3 and REM sleep stage and wake shifts (abbreviated as N1, N2, N3, REM and W) and their proportions of SSS (abbreviated as N1/SSS, N2/SSS, N3/SSS, REM/SSS and W/SSS). Results Compared with control group, WASO, PLMs, PLMs index of REM sleep, SSS, SSS/H and N2 were significantly increased in TLE group. Moreover, compared with control group, SpO2 was decreased in TLE group (P<0.05). Conclusion Our results suggest that TLE patients have sleep disorder manifested as disorder of sleep structure, increased incidents of respiratory and motion events.

The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.
Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Chromogenic Compounds ; Female ; Gene Expression Profiling ; methods ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization, Fluorescence ; methods ; Neoplasm Invasiveness ; pathology ; physiopathology ; Receptor, ErbB-2 ; genetics ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity

Objective To investigate the incidence of nutritional risk in infants with lower respiratory tract infection, and to compare the effects of different nutritional risks on clinical outcomes, and to provide evi-dence for clinical nutritional management of infantile lower respiratory tract infection. Methods Infants and young children with lower respiratory tract infection who were hospitalized in our hospital from January 2013 to March 2016 were selected as subjects. Nutritional risk screening was performed using the Nutritional Status and Growth Risk Screening Tool ( STRONGkids) . Results A total of 957 infants with lower respiratory tract infec-tions were included in the study. The incidence of high nutrition risk and low and medium nutritional risk were 17. 6％ and 82. 4％, respectively. The clinical cure rate was 68. 5％ and 71. 4％ respectively. The children with pneumonia and bronchitis had high nutritional risk. The incidence rates were 20. 60％ and 4. 87％, respectively, and the difference was statistically significant (χ2=25. 52, P=0. 000) . Time-effect single factor analysis ( Kaplan-Meier method):The hospitalization time for infants with low nutritional risk and high nutri-tional risk was 9. 3 ( 0. 3) d and 13. 3 ( 1. 0) d, respectively. The difference between the two groups was sta-tistically significant. (χ2=28. 33, P=0. 000) , the total hospitalization expenses were 5653. 5 ( 224. 8) yuan and 10079. 5 ( 1755. 8) yuan respectively. The difference between the two groups was statistically significant (χ2=4. 47, P=0. 034) . Multivariate COX regression analysis:High nutritional risk was a risk factor for hospi-talization of hospitalized infants with lower respiratory tract infection ( RR=1. 57, P=0. 024 ) . Conclusion There is a high incidence of high nutritional risk in infants with lower respiratory tract infection. Compared with children with low and moderate nutritional risk, the hospitalization time is longer, the hospitalization cost is in-creased, and the clinical cure rate is lower, which is the risk of clinical outcome. factor. Therefore, it is neces-sary to conduct nutrition risk screening for infants with lower respiratory tract infections, and provide a theoreti-cal basis for clinical nutrition evaluation and nutritional intervention.

OBJECTIVE: To express and purify the extra cellular full-length human apolipoprotein M(ApoM). METHODS: The ApoM gene fragment was amplified from the human liver cDNA library by PCR. The resulting product was cloned into pGEXT vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid E.coli JM109 and then transformed into E.coli DL21(DE3) where it was induced to express protein by IPTG. RESULTS: The ApoM gene was cloned by PCR and a 560 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoM gene in GenBank. Then ApoM cDNA gene fragment was induced by IPTG, and a 24 kD recombinant ApoM protein was tested on SDS-PAGE. CONCLUSION Human ApoM gene is successfully cloned and its recombinant proteins are expressed.
Apolipoproteins ; biosynthesis ; genetics ; isolation & purification ; Apolipoproteins M ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Lipocalins ; Molecular Sequence Data ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification