Study on the mechanism of Wuzi-Yanzong-Wan-medicated serum interfering with the mitochondrial permeability transition pore in the GC-2 cell induced by atractyloside / 中国天然药物

Wuzi-Yanzong-Wan (WZYZW) is a classic prescription for male infertility. Our previous investigation has demonstrated that it can inhibit sperm apoptosis via affecting mitochondria, but the underlying mechanisms are unclear. The purpose of the present study was to explore the actions of WZYZW on mitochondrial permeability transition pore (mPTP) in mouse spermatocyte cell line (GC-2 cells) opened by atractyloside (ATR). At first, WZYZW-medicated serum was prepared from rats following oral administration of WZYZW for 7 days. GC-2 cells were divided into control group, model group, positive group, as well as 5%, 10%, 15% WZYZW-medicated serum group. Cyclosporine A (CsA) was used as a positive control. 50 μmol·L-1 ATR was added after drugs incubation. Cell viability was assessed using CCK-8. Apoptosis was detected using flow cytometry and TUNEL method. The opening of mPTP and mitochondrial membrane potential (MMP) were detected by Calcein AM and JC-1 fluorescent probe respectively. The mRNA and protein levels of voltage-dependent anion channel 1 (VDAC1), cyclophilin D (CypD), adenine nucleotide translocator (ANT), cytochrome C (Cyt C), caspase 3, 9 were detected by RT-PCR (real time quantity PCR) and Western blotting respectively. The results demonstrated that mPTP of GC-2 cells was opened after 24 hours of ATR treatment, resulting in decreased MMP and increased apoptosis. Pre-protection with WZYZ-medicated serum and CsA inhibited the opening of mPTP of GC-2 cells induced by ATR associated with increased MMP and decreased apoptosis. Moreover, the results of RT-qPCR and WB suggested that WZYZW-medicated serum could significantly reduce the mRNA and protein levels of VDAC1 and CypD, Caspase-3, 9 and CytC, as well as a increased ratio of Bcl/Bax. However, ANT was not significantly affected. Therefore, these findings indicated that WZYZW inhibited mitochondrial mediated apoptosis by attenuating the opening of mPTP in GC-2 cells. WZYZW-medicated serum inhibited the expressions of VDAC1 and CypD and increased the expression of Bcl-2, which affected the opening of mPTP and exerted protective and anti-apoptotic effects on GC-2 cell induced by ATR.

Hyperoside protects mouse spermatocytes GC-2 cells from oxidative damage by activating the Keap1/Nrf2/HO-1 pathway / 南方医科大学学报

OBJECTIVE@#To study the protective effect of hyperoside (Hyp) against ydrogen peroxide (H2O2)- induced oxidative damage in mouse spermatocytes GC-2 cells and explore the role of the Keap1/Nrf2/HO-1 pathway in this protective mechanism.@*METHODS@#GC-2 cells were treated with 2.5 mmol/L azaacetylcysteine (NAC), 50, 100, and 200 μmol/L hyperoside, or the culture medium for 48 h before exposure to H2O2 (150 μmol/L) for 2 h. CCK-8 assay was used to detect the changes in cell viability, and cell apoptosis was analyzed using flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) activity and malondialdehyde (MDA) in the culture medium. Western blotting and RT-qPCR were used to detect the protein and mRNA expression levels of nuclear factor erythroid 2-related factor2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), and heme oxygenase-1 (HO-1); the nuclear translocation of Nrf2 was detected using immunofluorescence assay.@*RESULTS@#Exposure to H2O2 significantly lowered the proliferation rate, reduced the activities of SOD, GSH and CAT, and obviously increased MDA content, cell apoptosis rate, and the expressions of Keap1 and Nrf2 mRNA and Keap1 protein in GC-2 cells (P < 0.05 or 0.01). Treatment of the cells prior to H2O2 exposure with either NAC or 200 μmol/L hyperoside significantly increased the cell proliferation rate, enhanced the activities of SOD, GSH-PX and CAT, and lowered MDA content and cell apoptosis rate (P < 0.05). Treatment with 200 μmol/L hyperoside significantly decreased the mRNA and protein expressions of Keap1 and increased the expressions of HO-1 mRNA and the protein expressions of Nrf2 and HO-1 (P < 0.05 or 0.01). Hyperoside also caused obvious nuclear translocation of Nrf2 in the cells (P < 0.05).@*CONCLUSION@#Hyperoside protects GC-2 cells against H2O2- induced oxidative damage possibly by activation of the Keap1/Nrf2/HO-1 signaling pathway.

Effect of ligustrazine on Gas/cAMP/PKA signal activity and airway inflammation in asthmatic rats / 中国药理学通报

Aim To investigate the effect of ligus- trazine on G-protein coupled receptor signal activity in ovalbumin ( OVA)-sensitized and challenged asthmatic rats.Methods This study replicated an OVA-in¬duced juvenile asthmatic rat model, and investigated the effect of ligustrazine on airway hyperresponsive- ness, airway inflammation, lung histopathology, IL-4, IL-5 , 1L-13 , TNF-cx levels in serum, cAMP concentra¬tion in plasma, CREB, PKA, Gas and GRK2 expres¬sion in lung tissues.Results It was showed that the resistance of lung (RL) in asthmatic rats increased, while the pulmonary dynamic compliance ( Cdvn) sig¬nificantly decreased ( P < 0.05 ) compared with the control rats, and the lung tissue injury was serious; the levels of IL-4, IL-5, IL-13, and TNF-cx in serum were significantly increased ( P <0.01 ).The cAMP level in plasma was reduced and the CREB and PKA expres¬ sions were down-regulated significantly ( P < 0.05 ).The Gas expression in lung in asthmatic rats was down- regulated while the GRK2 expression was up-regulated ( P <0.01 ).Ligustrazine coulrl improve the lung inju¬ry and lung function of asthmatic rats, decrease RL and increase Cdvn ( P < 0.05 ) , decrease the levels of 1L-4, 1L-5, 1L-13, and TNF-a in serum (P <0.01) , increase cAMP in plasma and up-regulate the PKA and CREB expression, down-regulate the GRK2 expression and up-regulate the Gas expression significantly ( P < 0.01).Conclusions The above study shows that li¬gustrazine can reduce the AHR, inhibit the inflamma¬tion in asthma, and the up-regulating of Gas/cAMP/ PKA signal activity may be its mechanism.

Effect of Wuzi Yanzong Wan on Sperm Apoptosis by Intervening with Mitochondrial Permeability Transition Pore / 中国实验方剂学杂志

Objective:To study the effect of Wuzi Yanzong Wan on the expressions of voltage-dependent anion channel 1 （VDAC1）， adenine nucleotide transposase （ANT）， cyclophilin D （CypD） and other proteins， and analyze its mechanism in intervening with sperm mitochondrial permeability transition pore （mPTP） opening. Method:Forty rats were randomly divided into 4 groups， namely normal group， model group， positive group （Shengjing capsule， 1.6 g·kg-1·d-1）， and Wuzi Yanzong Wan group （4.0 g·kg-1·d-1）， with 10 rats in each group. Except for the normal group， tripterygium wilfordii glycosides （GTW， 30 mg·kg-1） was intragastrically administered for 8 weeks to establish the oligozoospermia model. After the 4th week， each group was given drugs through intragastric administration for 4 weeks， and fasted for 12 h after the last administration. These rats were anesthetized with 3% chloral hydrate， and their testis and epididymis tissues were collected. Western blot was used to determine the protein expressions of VDAC1，ANT，CypD，B-cell lymphoma/leukemia2 （Bcl-2）， Bcl-2 associated X protein （Bax）， Caspase-3， Caspase-9 in rat testis， Testicular tissue and its ultrastructure were observed under electron microscopy. The apoptosis in spermatogenic cells was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL）. Result:Western blot results showed that compared with the normal group， the expressions of VDAC1， CypD， Caspase-3， Caspase-9 and Bax/Bcl-2 in the model group were increased （P<0.01）. The expressions of VDAC1， CypD， Caspase-3， Caspase-9 and Bax/Bcl-2 were significantly decreased in the positive group and the Wuzi Yanzong Wan group compared with the model group （P<0.01）. There was no significant change in the expressions of ANT and Bcl-2 protein between the groups. Testicular ultrastructural evaluation showed different sizes and disordered arrangement of sperm mitochondria and a large number of swelling and vacuoles in the model group， while complete structure and neat arrangement of sperm mitochondria and much less swelling and vacuole in positive group and Wuzi Yanzong Wan group. TUNEL results showed that the apoptosis rate of spermatogenic cells in the model group was significantly higher than that of the normal group （P<0.01）， while the apoptosis rate in the positive drug and Wuzi Yanzong Wan group was significantly lower than that of the model group （P<0.01）. Conclusion:Wuzi Yanzong Wan may resist germ cell apoptosis by inhibiting the expressions of VDAC1， CypD and Bax， reducing the permeability of mPTP， and preventing the cascade activation reaction of the Caspase family of apoptosis proteins.

Role of T cells in acute kidney injury / 中国免疫学杂志

Acute kidney injury (AKI) refers to the clinical syndrome of rapid loss of renal function caused by various causes.AKI increases the risk of developing chronic kidney disease (CKD) and the mortality of patients.The increase in the rate has caused a great economic burden on the family and society.The pathogenesis of mediating AKI is numerous,and it is currently believed that innate and adaptive immune-mediated inflammatory reactions are involved in the initiation,progression and repair stage of AKI.T lymphocytes play a key role in it.This article will review the progress of the role of T cells in AKI.

Research progress of role of NLRP3 inflammasome in lupus nephritis / 中国免疫学杂志

Systemic lupus erythematosus (SLE)is an autoimmune disease whose pathogenesis is extremely complicated.With the further research,the role of inflammasome in the pathogenesis of Lupus nephritis has also been gradually emphasized.Among them,the nucleotide-binding oligomerization domain-like receptors family pyrin domain containing 3 (NLRP3) inflammasome is the most exhaustive inflammasome.We summarize the related studies on the role of NLRP3 inflammasome in Lupus nephritis in recent years.We found that NLRP3 inflammasome not only plays an important role in the pathogenesis of Lupus nephritis,but also participates in the process of kidney injury by circulating immune cells and renal innate cells.Finally,we introduced two specific inhibitors of NLRP3 inflammasome,β-hydroxybutyrate and MCC950,which provided a new strategy for the treatment of Lupus nephritis.

Herbs protect against osteoporosis through anti-inflammation action / 中国组织工程研究

BACKGROUND: The occurrence and development of osteoporosis are shown to be directly related to the inflammatory response induced by immune dysfunction. OBJECTIVE: To summarize the mechanisms of osteoclast differentiation and formation at cellular and molecular levels, as well as the underlying mechanisms of several kinds of medical herbs against osteoporosis, thus paving ways for finding more effective and safe herbs for anti-osteoporosis.METHODS: PubMed database was retrieved using the English keywords of (osteoporosis OR bone loss) AND lipopolysaccharide AND bone resorption", and WanFang database was searched with the Chinese keywords of "osteoarthritis, receptor activator of nuclear factor-κB ligand, lipopolysaccharide,Dendrobium moniliforme,Portulaca oleracea,Ampelopsis sinica,Schizonepeta".The literatures addressing osteoporosis, inflammation and herbal medicine were screened, and those using lipopolysaccharide-induced mouse models were included. Finally, four eligible literatures were enrolled for review. RESULTS AND CONCLUSION:In vivo experiments,CT images and pathological sections of the cancellous bone in the mouse distal femur show that Dendrobium moniliforme,Purslane oleracea,Ampelopsis sinica,and Schizonepeta exert inhibitory effects on lipopolysaccharide-induced osteoporosis.In vitro experiments reveal that these four kinds of herbs fight against osteoporosis by inhibiting osteoclast differentiation and further reducing bone resorption.

Protective effect of Wuziyanzong Pills on rats with experimental oligoasthenospermia and its action mechanism / 中华男科学杂志

<p><b>Objective</b>To investigate the protective effect of Wuziyanzong Pills (WYP) in the rat model of oligoasthenospermia (OAS) and its action mechanism.</p><p><b>METHODS</b>Sixty male SD rats were equally randomized into six groups: normal control, OAS model, Shengjing Capsules (1.6 g per kg of the body weight), low-dose WYP (1 g per kg of the body weight), medium-dose WYP (2 g per kg of the body weight), and high-dose WYP (4 g per kg of the body weight). The OAS model was established by intragastric administration of Tripterygium glucoside at 30 mg per g per d for 6 weeks. From the 3rd week of modeling, the rats of the medication groups were treated intragastrically with corresponding drugs for 4 weeks. Then all the rats were sacrificed for measurement of the testicular and epididymal organ coefficients, examination of epididymal sperm quality and apoptosis, and detection of the openness of the sperm mitochondrial permeability transition pore (MPTP). Histopathological changes in the testis were observed by HE staining and the apoptosis of spermatogenic cells determined by Hochest staining.</p><p><b>RESULTS</b>WYP obviously improved the organ coefficients of the testis and epididymis, increased sperm concentration, motility and viability, decreased the apoptosis of spermatogenic cells, and inhibited the abnormal openness of MPTP in the OAS model rats. HE staining showed that the number and levels of spermatogenic cells were significantly increased while Hochest staining manifested that the apoptosis of spermatogenic cells was remarkably inhibited in the seminiferous tubules of the testis in the WYP-treated rats.</p><p><b>CONCLUSIONS</b>WYP can improve sperm quality and reduce the apoptosis of spermatogenic cells (including sperm) in OAS model rats, which may be related with its inhibitory effect on the abnormal openness of MPTP.</p>

Puerarin prevents bone loss in ovariectomized mice and inhibits osteoclast formation in vitro / 中国天然药物

The present study aimed at investigating the effects of Puerarin (PR), a major isoflavonoid isolated from the Chinese medicinal herb Puerariae radix, on bone metabolism and the underlying mechanism of action. The in vivo assay, female mice were ovariectomized (OVX), and the OVX mice were fed with a diet containing low, middle, and high doses of PR (2, 4, and 8 mg·d(-1), respectively) or 17β-estradiol (E2, 0.03 μg·d(-1)) for 4 weeks. In OVX mice, the uterine weight declined, and intake of PR at any dose did not affect uterine weight, compared with the control. The total femoral bone mineral density (BMD) was significantly reduced by OVX, which was reversed by intake of the diet with PR at any dose, especially at the low dose. In the in vitro assay, RAW264.7 cells were used for studying the direct effect of PR on the formation of osteoclasts. PR reduced the formation of tartrate resistant acid phosphatase (TRAP)-positive multi-nucleated cells in the RAW 264.7 cells induced by receptor activator for nuclear factor-κB Ligand (RANKL). MC3T3-E1 cells were used for studying the effects of PR on the expression of osteoprotegerin (OPG) and RANKL mRNA expression in osteoblasts. The expression of OPG mRNA and RANKL mRNA was detected by RT-PCR on Days of 5, 7, 10, and 12 after PR exposure. PR time-dependently enhanced the expression of OPG mRNA and reduced the expression of RANKL mRNA in MC3T3-E1 cells. In conclusion, our results suggest that PR can effectively prevent bone loss in OVX mice without any hyperplastic effect on the uterus, and the antiosteoporosis activity of PR may be related to its effects on the formation of osteoclasts and the expression of RANKL OPG in osteoblasts.

Bone metabolism and gut microbiota / 中国组织工程研究

BACKGROUND:The gut microbiota in our intestine performs numerous useful functions and has a major impact on the host’s health. Recently some studies have revealed that the gut microbiota cannot only control intestinal activity but also affect bone metabolism by regulating the immune system. OBJECTIVE:To review the new research development in the effects of gut microbiota on bone metabolism. METHODS: We retrieved the PubMed database using “gut microbiota, immune system, bone metabolism, osteoporosis” as keywords. A total of 46 articles were included which were related to gut microbiota, immune system and bone metabolism. For the articles in the same field, those published recently or in authorized journals were selected. RESULTS AND CONCLUSION:Gut microbiota-osteoporosis research wil bridge the gaps between bone physiology, gastroenterology, immunology, and microbiology.In vivo experiments in the germ-free mice and human body have found that the gut microbiota has important effects on bone metabolism, and the intervention of antibiotics, probiotics and prebiotics has further confirmed the effects of gut microbiota on bone mass. The gut microbiota has more obvious effects on bone mass in the adolescent and post-menopause periods.

Puerarin effects on the mRNA expression of osteoblast differentiation-related proteins / 中国组织工程研究

BACKGROUND:Experimental studies have showed that puerarin has an obvious protective effect on osteoporosis in ovariectomized and orchiectomized mice. But the influence of puerarin in the molecular level in the process of osteoblast differentiation is seldom reported. OBJECTIVE:To observe the effect of puerarin on the mRNA expression of alkaline phosphatase, bone sialoprotein, osteopontin and osteocalcin in osteoblasts. METHODS:The MC3T3-E1 cells from mice cultured in vitro were randomly divided into control group, puerarin group (10-6 mol/L puerarin) and estradiol group (10-7 mol/L estradiol) to observe the effects of puerarin on the differentiation of osteoblasts. mRNA expression of alkaline phosphatase, bone sialoprotein, osteopontin and osteocalcin in MC3T3-E1 cells was determined using RT-PCR method. RESULTS AND CONCLUSION:Puerarin and estradiol both could prolong the expression of alkaline phosphatase that reached the peak at 12 days. Puerarin and estradiol strengthened the mRNA expression of bone sialoprotein at 10 and 12 days, reduced expression of osteopontin at 5 and 12 days, and increased expression of osteocalcin at 10 and 12 days. These results reveal that puerarin can induce the differentiation of cultured osteoblasts by influencing osteoblast differentiation-related protein mRNA expressions, which may be one of the important molecular mechanisms of puerarin for prevention of osteoporosis.

Wuzi yanzong pills increases sperm mitochondrial membrane potential and protects its ultrastructure in oligo-asthenozoospermia model rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effects of Wuzi Yanzong Pills (WYP) on sperm mitochondrial membrane potential (MMP) and its ultrastructure in oligo-asthenozoospermia model rats.</p><p><b>METHODS</b>Oligo-asthenozoospermia models were made in 50 male rats weighing 200 - 220 g by intragastric administration of Tripterygium Glucosides at 30 mg per kg per d for 8 weeks, and then equally allocated to a model control, a Huangjing Zanyu Capsule (HZC) control, a low-dose WYP, a medium-dose WYP, and a high-dose WYP group. Another 10 age-matched normal male rats were included as normal controls. The rats in the model and normal control groups were given intragastrically distilled water at 10 ml/kg, those in the HZC group administered HZC at 3.01 g/kg, and those in the low-, medium- and high-dose WYP groups medicated with WYP at 2.30, 4.60 and 9.20 g/kg, respectively, once daily for 30 days. At 30 minutes after the last administration, we detected the sperm MMP by JC-1 fluorescent staining and flow cytometry, and examined the sperm ultrastructure under the JEM-1230 transmission electron microscope.</p><p><b>RESULTS</b>JC-1 + % and its fluorescence intensity were (33.77 +/- 6.19)% and 1 468 +/- 496 in the model control, (56.34 +/- 10.35)% and 3 277 +/- 895 in the HZC control, (40.80 +/- 10.40)% and 2 016 +/- 767 in the low-dose WYP, (59.40 +/- 6.51)% and 3 897 +/- 643 in the medium-dose WYP, and (60.71 +/- 7.81)% and 3 371 +/- 647 in the high-dose WYP group, significantly reduced in comparison with (70.80 +/- 4.92)% and 4 360 +/- 945 in the normal control group (P < 0.05), but remarkably higher in the medium- and high-dose WYP groups than in the model controls (P < 0. 05). After modeling, the sperm membrane was loose and degenerated, the mitochondria swelling, variously sized and with incomplete membrane, and the axonemal structure unclear or ruptured. After 30 days of WYP administration, compared with the model control group, the rats exhibited integrated sperm membrane and mitochondrial membrane, reduced mitochondrial swelling and basically normal axonemal and microtubular structures.</p><p><b>CONCLUSION</b>Tripterygium Glucosides could decrease the sperm mitochondrial membrane potential and damage the mitochondrial structure, while WYP could significantly increase the sperm mitochondrial membrane potential and reduce the sperm mitochondrial structure damage. The protection of the integrity of sperm mitochondrial structure and function is one of the mechanisms of WYP acting on oligo-asthenozoospermia.</p>

Distribution of genotypes in ESBLs producing E. coli strains isolated from posthepatitic cirrhosis' patients with bloodstream infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.</p><p><b>METHODS</b>E. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.</p><p><b>RESULTS</b>A total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.</p><p><b>CONCLUSION</b>CTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.</p>

Establishment of a quantitative detection method for Golgi protein73 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.</p><p><b>RESULTS</b>The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.</p><p><b>CONCLUSION</b>Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.</p>

Effects of epithelial cell injury of the lower respiratory tract in the pathogenesis of allergic responses in a rat model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Asthma is a complex disease involving genetic and environment interactions. Atopy is a strong risk factor for asthma. The airway epithelium not only forms a physical barrier but also provides immune defense against harmful materials. To explore the effects of airway epithelium on asthma, we hypothesized that environmental injuries could act on bronchial epithelial cells and damage the physical barrier, which might facilitate allergens to stimulate immunoreactions and play an important role in the pathogenesis of asthma.</p><p><b>METHODS</b>Thirty eight-week-old male Wistar rats were randomly divided into five groups with six rats in each group: control group, asthma group, ovalbumin (OVA) + OVA group, lipopolysaccharide (LPS) group and LPS + OVA group. In the control group, 0.9% saline was injected intraperitoneally on day 1. Fourteen days later, the rats were exposed to aerosolized 0.9% saline. In the asthma group, the rats were sensitized with an injection of 10 mg of OVA, followed by an aerosolized 2% OVA challenge 14 days later. The OVA + OVA group was sensitized by an inhalation 2% OVA, 20 minutes a day, from day 1 to day 7, and then OVA challenged in the same way as the asthma group. In the LPS group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with saline as in the control group. While in the LPS + OVA group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with OVA as in the asthma group. The expression of interleukin (IL)-4, interferon-gamma (IFN-γ) and thymic stromal lymphopoietin (TSLP) in the lungs was detected by reverse transcription polymerase chain reaction (RT-PCR) and the pulmonary pathological changes were also observed. The level of IL-4, IFN-γ and IgE in plasma was detected by enzyme-linked immunosorbent assay (ELISA). Bronchoalveolar lavage fluid (BALF) was collected to conduct differential cell counts. Flow cytometry analysis was also used to count Th1 and Th2 cells.</p><p><b>RESULTS</b>The pathological changes in the LPS + OVA group were similar to the asthma group, while in other groups, the pathological changes were not obvious. The ratio of lymphocytes in BALF, IL-4/IFN-γ in plasma and the expression of the TSLP and IL-4 in the asthma and LPS + OVA groups were higher than in the control group and the OVA + OVA group (P < 0.05). The level of IgE was higher in the asthma, LPS and LPS + OVA groups than in the control group and the OVA + OVA group (P < 0.05). By flow cytometry analysis, the Th1/Th2 ratio was lower in the LPS + OVA and asthma groups than in other groups (P < 0.05).</p><p><b>CONCLUSIONS</b>The experiment results show that the injury to the bronchial epithelial layer may be the initial event of allergic responses. This finding implies that a rational approach to therapeutics would be to increase the resistance of the airways to environmental injuries rather than concentrating on suppressing inflammation.</p>

A clinical and pathological analysis of 22 cases of primary sclerosing cholangitis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To characterize the clinical, laboratory, imaging and pathological features of primary sclerosing cholangitis (PSC) and investigate the impact of ursodeoxycholic acid (UDCA) therapy on patient prognosis.</p><p><b>METHODS</b>The medical records of 22 patients diagnosed with PSC between 2002 and 2011 were retrospectively reviewed. The PSC diagnosis had been made in patients with suspect biochemical abnormalities following evaluation by magnetic resonance cholangiopancreatography (MRCP) and/or endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC). Fibrosis and inflammation were assessed by immunohistochemical analyses of tissue biopsies. Outcome of patients treated with UDCA (13-15 mg/kg/day, oral) were compared to that of patients without UDCA treatment by the X2 or corrected X2 tests.</p><p><b>RESULTS</b>Among the 22 PSC patients, the majority was male (n=15) and presented with fatigue, dark urine, and body weight loss (n=15). Four cases had ulcerative colitis. At admission, all 22 cases showed elevated levels of alkaline phosphatase[ALP: (348+/-184) U/L], 19 cases showed elevated alanine aminotransferase [ALT: (94.0+/-67.0) U/L] and aspartate aminotransferase [AST: (98.0+/-67.0) U/L], and 15 cases showed elevated levels of total bilirubin (99.0+/-115.0) mumol/L and direct bilirubin (74.4+/-92.4 mumol/L. ERCP examination showed segmental intrahepatic bile duct stenosis with expansion, and stiff and enlarged gallbladder bile ducts, but unclear findings for the common bile ducts and pancreatic ducts. MRCP showed beading of the intrahepatic bile duct, stiffness of the bile duct wall, and dilation of the common bile duct. Fibrosis and inflammation were observed in the bile ducts, along with hyperplasia and the typical features of "onion skin" fibrosis and fibrous obliterative cholangitis. Five of the 10 patients treated with UDCA improved, and seven of the 12 patients in the non-UDCA treatment group improved. There was no statistically significant difference in outcome between the groups (paired X2=0.333, corrected X2=0.083, P more than 0.05).</p><p><b>CONCLUSION</b>PSC patients were predominantly male and the common clinical manifestations were fatigue, dark urine, and body weight loss. At admission, serum biochemical indicators of cholangitis were increased significantly and subsequent imaging studies confirmed the suspected diagnosis by showing obvious characteristic changes. UDCA treatment did not significantly improve patient prognosis.</p>

Efficacy evaluation of laparoscopic gastric bypass for the treatment of obese type 2 diabetes mellitus / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the treatment outcomes of obese type 2 diabetes mellitus (T2DM) after laparoscopic gastric bypass.</p><p><b>METHODS</b>The clinical data of 18 patients with obese T2DM who underwent laparoscopic Roux-en-Y gastric bypass in Beijing Shijitan Hospital between March 2009 and February 2011 were retrospectively analyzed. The clinical parameters included preoperative and postoperative blood glucose, blood lipid, nutrition status and weight lose.</p><p><b>RESULTS</b>Eighteen patients included 8 men and 10 women. The range of age was 27-62 years (mean, 42.4±10.7 years). The range of BMI was 28.7-57.4 kg/m(2)(mean, 34.9±6.9 kg/m(2)). All the patients underwent laparoscopic Roux-en-Y gastric bypass, and no mortality, complication or conversion to open operation occurred. At 3 months after operation, there were significant changes in OGTT, BMI, HbA1c, Homa-IR and Homa-β(all P<0.05). Fourteen patients(77.8%) showed clinical complete remission, and the overall effective rate was 100%(18/18). The level of blood lipid decreased significantly (P<0.05), and the change of nutritional status was not statistically significant(P>0.05).</p><p><b>CONCLUSION</b>Gastric bypass is an effective treatment for obese type 2 diabetes mellitus.</p>

The verification of a new DNA double-detection diagnostic kit for identifying novel influenza A (H1N1) / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To verify a new kit of "universal and novel influenza A (H1N1) virus nucleic acid double-detection methods (PCR-fluorescence probe)".</p><p><b>METHODS</b>150 cases of throat swab specimens were collected consecutively. After RNA was extracted, the specimens were detected by the verified kit. At the same time, the same specimens were detected by Real-time PCR diagnostic kit from Beijing CDC as the control. The data were analysed by the Kappa in agreement and by McNemar chi2 in difference test.</p><p><b>RESULTS</b>The consistency rate of the verified kit and the Beijing CDC kit was universal primer M 97.33%, H1N1 98.67% respectively. The Kappa test and McNemar chi2 test showed that two methods had a higher consistency. Compared to the CDC kit, the "false negative rate" and "false-positive rate"of double-check kit were lower.</p><p><b>CONCLUSION</b>The kit of "universal and novel influenza A (H1N1) virus nucleic acid double-detection methods (PCR-fluorescence probe)" from Shanghai Kehua Bio-Engineering Co., Ltd can be used to detect influenza A and novel influenza A (H1N1).</p>

The early diagnosis value of EV 71 IgM class antibodies in the hand, foot and mouth disease / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Assessment of detection of IgM antibodies for human enterovirus 71 (EV 71) in early diagnosis for the hand, foot and mouth disease (HFMD).</p><p><b>METHOD</b>The sera and throat swabs from 38 patients which were clinical diagnosis as HFMD, were continuous daily collected in our hospital in 2010. These specimens were detected by EV 71 IgM antibodies assay, real time RT-PCR methods for EV 71 and Enterovirus.</p><p><b>RESULTS</b>Among 38 HFMD patients, the cumulative positive rates of EV 71 IgM antibodies were: 60.5% on day 1, 71.1% on day 2, 81.5% in the first 3-4 days, 92.1% on day 5, 92.1% on day 6, and the positive rate of nucleic acid detected by the real time RT-PCR for EV 71 and Enterovirus were 60.5%, 73.6%.</p><p><b>CONCLUSION</b>The positive rate of EV 71 IgM antibodies in the hand, foot and mouth disease just can occur on day 1, and reach to peak on day 5, which can be used as one of indicators of early diagnosis of hand, foot and mouth disease.</p>

Clinical and laboratory characteristics of a new influenza A (HIN1) epidemic / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analysis the clinical and laboratory characteristics of Patients infected with new influenza A (HIN1) virus.</p><p><b>METHODS</b>All cases with new influenza A (H1N1) confirmed on polymerase chain reaction assay on throat swabs. There were included in a prospective evaluation of clinical characteristics, laboratory results, treatment and overcome of new influenza A (H1N1).</p><p><b>RESULTS</b>There were 35 patients in the epidemic. Clinical illness developed within a mean of 1.7 days. Fever occurred in 97.1%, sore throat 65.7% cough 51.4%, headache 28.6%, and myalgia 31.4%. All patients were treated with oseltamivir lasted 5 days. The mean duration of viral shedding was 4.5 days. All were cured and left hospital after day 7.</p><p><b>CONCLUSION</b>It was infected by new influenza A (H1N1) typically in this epidemic.</p>