BACKGROUND: Fleece Flower Root Granule, which consists of Fleece Flower Root, Desert-living Root, Achyranthes Root, and so on,possesses the effects of nourishing the liver and kidney, supplementing essence and blood, and prolonging life.OBJECTIVE: To investigate the effect of. Fleece Flower Root Granule, a Chinese remedy, on the content of serum testosterone and the expression of proliferating cell nuclear antigen of testicular spermatogenic cell in subacute aging rat induced by D-galactose.DESIGN: A controlled study with completely randomized grouping design.SETTING: Institute of Basic Medicine of Chengde Medical College.MATERIALS: The experiment was conducted in Institute of Basic Medicine of Chengde Medical College from April to July 2003. Totally 32 adult male SD rats were at random divided into 4 groups: normal control group, model group, drug group and negative control group,with 8 in each group.METHODS: [1] The rats in model group, drug group and negative control group were intraperitoneally injected D-galactose, 200 mg/kg ,once a day for consecutive 40 days to establish subacute aging rat models. After model was set up, no treatment was given further for rats in model group; while rats in drug group were intragastrically given Fleece Flower Root Granule (The granule was made from sex Chinese medicinal herbs of Fleece Flower Root, Achyranthes, Root Desertliving Root, Poria, Red Sage Root and Epimedium, the concentration was 1.5 g/L in raw drug after decoction and preparation); rats in negative group were intragastrically given normal saline, in the same volume as that of the Granule and the time as the same with that for the rats in drug group. Rats in normal control group, with no model establishment, were given the same intervention with that for rats in negative group. [2]Enzyme-linked immunosorbent assays (ELISA) was used for the level of serum testosterone, and inmmunohistochemistry was used for the condition of expression of proliferating cell nuclear antigen, with the karyon in orange brawn yellow being taken as positive. [3]Measurement difference was analyzed by t-test.MAIN OUTCOME MEASURES: The content of serum testosterone and the expression of proliferating cell nuclear antigen of testicular spermatogenic cell of rats in each group.RESULTS: All 32 rats entered the result analysis. [1] The level of serum testosterone:the level of testosterone in model group was obviously lower than that in normal control group [(0.52±0.14),(1.26±0.32)μg/L, P＜ 0.05]; the level in drug group was obviously higher than that in model group [(1.16±0.32)μg/L, P ＜ 0.05]. [2] The positive rate of testicular proliferating cell nuclear antigen in rats: The rate in model group was obviously lower than that in normal control group [(23.26±3.38)%,(39.15±3.48)%, P ＜ 0.05]; and the rate in drug group was obviously higher than that in model group [(37.47±3.18)%, P ＜ 0.05]. [3]rhe distribution and change of proliferating cell nuclear antigen in seminiferous tubule: The positive staining of proliferating cell nuclear antigen was found in slices of all groups, the immunological positive products were located at karyon, usually in brown yellow granules, only a small part of them in diffuse state; the immunological positive cells were mainly the spermospores, besides a small quantity were spermiocyte, supporting the cell and myoid cell nuclear to be stained.CONCLUSION: Fleece Flower Root Granule can increase the content of serum testosterone and reduce the level of expression of proliferating cell nuclear antigen of testicular spermatogenic cell in aging rat, thus delaying degeneration of genital function, possessing a certain role in delaying aging.

Objective To investigate the effects of inhaling high concentration of hydrogen gas on the expressions of endoplasmic reticulum stress(ERS) related protein glucose regulated protein 78 (GRP78),Caspase-12 and the neural cell apoptosis and related proteins Bcl-2 and Bax in the rats with focal cerebral ischemia reperfusion(I/R) injury.Methods Seventy-two healthy SPF male Sprague-Dawley rats were selected and then randomly divided into the control group(Ⅰ:without any treatment),sham operation group (Ⅱ),cerebral IRI group (Ⅲ) and hydrogen gas treatment group (Ⅳ),18 cases in each group.Focal cerebral ischemia reperfusion injury (IRI) model was induced by using the suture-occluded method.The neurological deficits score (NDS) was assessed at 24 h after cerebral reperfusion in four groups.The cerebral infarction severity and size were detected by TTC staining and neuronal apoptosis of brain cortex were tested by TUNEL technique.The apoptosis index (AI) was calculated.Then the expressions of GRP78,Caspase-12,Bcl-2 and Bax were assessed by Western blot and immunohistochemistry.Results As compared with the group Ⅰ and Ⅱ,NDS score,cerebral infarction size,AI and the expressions of GRP78,Caspase-12 and Bax in cerebral cortex in the group Ⅲl and Ⅳ were significantly increased,while the expression of Bcl-2 in cerebral cortex was markedly decreased(P＜0.05);compared with the group Ⅲ,NDS score,brain infarction size,AI and the expression of Caspase-12 and Bax in cerebral cortex in the group Ⅳ were markedly decreased,while the expressions of GRP78 and Bcl-2 were dramatically increased (P＜0.05).Conclusion Inhaling high concentration of hydrogen gas has a certain protective effect on cerebral IRI in rats through increasing endoplasmic reticulum GRP78 protein expression after IRI and inhibiting Caspase-12 activation,thus inhibiting ERS and promoting the repair function of endoplasmic reticulum.

Objective To investigate the role of phosphatidylinositol 3?kinase∕protein kinase B∕glycogen synthase kinase?3β ( PI3K∕Akt∕GSK?3β) signaling pathway in hydrogen?induced inhibition of neuronal ap?optosis induced by focal cerebral ischemia?reperfusion ( I∕R) in rats. Methods One hundred and twenty
healthy adult male Sprague?Dawley rats, weighing 200-220 g, were divided into 5 groups ( n=24 each) u?sing a random number table: sham operation group ( group S); cerebral I∕R group ( group I∕R); hydrogen group (group H2); LY294002 (specific PI3K inhibitor) group (group LY); LY294002+hydrogen group ( group LY+H2 ) . Focal cerebral I∕R was induced by occlusion of the middle cerebral artery for 1 h followed by 24 h reperfusion. In H2 and H2+LY groups, the animals inhaled 67% H2+33% O2 for 2 h starting from onset of reperfusion, and then inhaled H2 for 2 h every 6 h. In LY and LY+ H2 groups, LY294002 ( 10 mmol∕L) 10 μl was injected into the lateral cerebral ventricle at 10 min before reperfusion. Neurologic defi?cit was evaluated and scored ( NDS) at 24 h of reperfusion. The rats were then sacrificed, and the brains were removed for measurement of the cerebral infarct size ( by TTC staining) and apoptosis in cortical neu?rons ( by TUNEL) and for determination of the expression of Akt in the ischemic cerebral cortex, phospho?rylated Akt ( p?Akt) , GSK?3β and phosphorylated GSK?3β ( p?GSK?3β) and Bcl?2 and Bax positive cell count in the ischemic cerebral cortex ( by immuno?histochemistry) . The apoptosis index ( AI) , p?Akt∕Akt ratio and p?GSK?3β∕GSK3?β ratio were calculated. Results Compared with group S, the NDS, cerebral infarct size, AI, p?Akt∕Akt ratio, p?GSK?3β∕GSK?3β ratio and Bax positive cell count were significantly increased, and the Bcl?2 positive cell count was significantly decreased in group I∕R ( P<0?05) . Compared with group I∕R, the NDS, cerebral infarct size, AI and Bax positive cell count were significantly de?creased, and the p?Akt∕Akt ratio, p?GSK?3β∕GSK?3β ratio and Bcl?2 positive cell count were significantly increased in group H2 ( P <0?05) , and no significant changes were found in the parameters mentioned a?bove in LY and LY+H2 groups ( P>0?05) . Compared with group H2 , the NDS, cerebral infarct size, AI and Bax positive cell count were significantly increased, and the p?Akt∕Akt ratio, p?GSK?3β∕GSK?3βratio and Bcl?2 positive cell count were significantly decreased in group LY+H2 ( P<0?05) . Conclusion The mechanism by which hydrogen inhibits focal cerebral I∕R?induced neuronal apoptosis is associated with the activation of PI3K∕Akt∕GSK?3β signaling pathway in rats.

Objective To investigate the effect of high concentration hydrogen gas on neurons in the rat hippocampus CA1 region during global cerebral ischemic/reperfusion injury (GCIR) Methods Four-vessel occlusion was used to establish rat model with GCIR injury. One hundred and five healthy male Sprague-Dawley rats were randomly divided into sham operation group(SH group, n = 15), model group(4-VO group, n = 45) and treatment group(4-VO+H2 group,n = 45). After 72 h and 9 d reperfusion, hippocampal CA1 region pyramidal neurons in every group were detected with Nissle staining , immunohistochemical neuron-specific nuclear protein (NeuN), specific protein antibody microglial cells (Iba1) staining and the relationship of position between neurons and microglia was observed through fluorescence double staining. We used Morris water maze to test the space orientation ability and the learning and memory ability in rats after 9 d reperfusion. Results Compared with those of 4-VO group,the neurons of hippocampus CA1 region were closer to normal in 72 h and 9 d in 4-VO+H2 group and neuron form and the number of neuron survival were increased significantly (P < 0.05);immunohistochemical staining showed that the number of neuron survival in 4-VO+H 2 group was obviously higher than that in 4-VO group (P < 0.05) and the number of microglia in 4-VO group was obviously higher than that in 4-VO+H2 group (P < 0.05). Water maze experiment showed that the swimming time in quadrant Ⅳ in 4-VO+H2 group was longer than that in 4-VO group (P < 0.05). Conclusion Inhalation of high concentration hydrogen gas has prominent protective effect on neurons of rat hippocampal CA1 region during reperfusion. The mechanism may be related with inhibiting the microglia excitation and activation during GCIR.

Objective To explore the protective effect of in-taking high concentration hydrogen gas on neurons and dendritic spines in hippocampus CA1 region of rats after globe cerebral ischemia/reperfusion (I/R) injury and its mechanism.Methods Four-vessel occlusion (4VO) was used to establish the models of global cerebral I/R injury in rats.One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into 3 groups using a random number table:sham-operated group (inhaled 67％ N2 and 67％ O2,n=40),model group (inhaled 67％ N2 and 67％ O2 during reperfusion,n=40),and treatment group (inhaled 67％ H2 and 67％ O2 during reperfusion,n=40).After 72 h,5 and 9 d reperfusion,neuron-specific nuclear (NeuN) protein expression in the pyramidal neurons of the hippocampal CA1 region was detected with immumohistochemical staining and the positive cells were counted.And the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum were tested with colorimetry.Water maze test was used to measure the spatial orientation and memory function and Golgi staining to detect the number of dendritic spines in neurons 9 d after reperfusion.Results (1) Immunohistochemical staining of NeuN results showed that as compared with those in the model group,the neurons ofhippocampus CA1 region were significantly closer to normal with relatively intact structure,and the number of positive neurons was significantly increased in the treatment group 72 h,5 d,and 9 d after reperfusion (P＜0.05).With the reperfusion time being prolonged,the number of NeuN stained positive neurons at different time points of reperfusion in model group was gradually decreased (P＜0.05),and the numeric of the NeuN stained positive neurons at different time points of reperfusion in treatment group was slightly declined without significant difference (P＞0.05).(2) The serum SOD activity in the treatment group was significantly higher than that in the model group and sham-operated group (P＜0.05),while the MDA content in the treatment group was significantly lower than that in model group 72 h,5 d,and 9 d after reperfusion (P＜0.05).And with the reperfusion time being prolonged,the SOD activity at different time points of reperfusion showed no significant difference among the difference groups (P＞0.05).But the MDA content at different time points ofreperfusion between model group and treatment group was significantly different (P＜0.05);with the reperfusion time being prolonged,the MDA content was gradually decreased in both groups.(3) Nine d after reperfusion,water maze test found that the incubation period of treatment group was significantly shorter than that of model group (P＜0.05);the IV quadrant swimming time of space exploration in the treatment group was significantly longer than that in the model group (P＜0.05).(4) Golgi staining showed that the complexity of the neuronal dendrites branch and the number of dendritic spines of neurons in the hippocampal CA1 region of treatment group were increased than those in the model group;high-power oil microscopy indicated that the density of dendritic spines in the treatment group was significantly higher than that in the model group (P＜0.05).Conclusion In-taking of high concentrations hydrogen gas during reperfusion can definitely protect neurons in hippocampal CA1 region after globe cerebral I/R injury,and improve learning and memory function,whose mechanism may be related to hydrogen protecting the structure and function of neurons and dendritic spines,and inhibiting oxidative stress to reduce oxidative damage.

Background: Dysimmunity plays a key role in diabetes, especially type 1 diabetes mellitus. Islet-specific autoantibodies (ISAs) have been used as diagnostic markers for different phenotypic classifications of diabetes. This study was aimed to explore the relationships between ISA titers and the clinical characteristics of diabetic patients. Methods: A total of 509 diabetic patients admitted to Department of Endocrinology and Metabolism at the Affiliated Hospital of Nantong University were recruited. Anthropometric parameters, serum biochemical index, glycosylated hemoglobin, urinary microalbumin/creatinine ratio, ISAs, fat mass, and islet β-cell function were measured. Multiple linear regression analysis was performed to identify relationships between ISA titers and clinical characteristics. Results: Compared with autoantibody negative group, blood pressure, weight, total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), visceral fat mass, fasting C-peptide (FCP), 120 minutes C-peptide (120minCP) and area under C-peptide curve (AUCCP) of patients in either autoantibody positive or glutamate decarboxylase antibody (GADA) positive group were lower.Body mass index (BMI), waist circumference, triglycerides (TGs), body fat mass of patients in either autoantibody positive group were lower than autoantibody negative group. GADA titer negatively correlated with TC, LDL-C, FCP, 120minCP, and AUCCP.The islet cell antibody and insulin autoantibody titers both negatively correlated with body weight, BMI, TC, TG, and LDL-C. After adjusting confounders, multiple linear regression analysis showed that LDL-C and FCP negatively correlated with GADA titer. Conclusion Diabetic patients with a high ISA titer, especially GADA titer, have worse islet β-cell function, but less abdominal obesity and fewer features of the metabolic syndrome.

Dysimmunity plays a key role in diabetes, especially type 1 diabetes mellitus. Islet-specific autoantibodies (ISAs) have been used as diagnostic markers for different phenotypic classifications of diabetes. This study was aimed to explore the relationships between ISA titers and the clinical characteristics of diabetic patients.

Dysimmunity plays a key role in diabetes, especially type 1 diabetes mellitus. Islet-specific autoantibodies (ISAs) have been used as diagnostic markers for different phenotypic classifications of diabetes. This study was aimed to explore the relationships between ISA titers and the clinical characteristics of diabetic patients.

Objective To investigate the relationship between prognostic nutritional index (PNI) and neutropenia after adjuvant chemotherapy in patients with colorectal cancer. Methods The clinical data of 44 patients with colorectal cancer performed adjuvant chemotherapy in Shunyi District Hospital from December 2014 to January 2018 were retrospectively analyzed, and the patients were divided into group A (grade 0-2 neutropenia) and group B (grade3-4 neutropenia) according to the degree of neutropenia. The serum albumin, peripheral lymphocyte counts, and neutrophil counts within 1 week before chemotherapy were collected, and the PNI was calculated. The chi-square test and rank sum test were used to compare the clinical data, body mass index (BMI), baseline neutrophil count, and PNI between the two groups. Logistic regression analysis was used to analyze the risk factors for neutropenia after chemotherapy. Results The baseline median neutrophil counts and median PNI in group A were 3.17×109/L [(1.38-7.79)×109/L] and 50.40 (37.40-57.05), and in group B were 2.54 ×109/L [(1.22-3.87) ×109/L] and 45.50 (37.95-50.95). The baseline neutrophil counts and PNI in group A were significantly higher than those in group B, the differences between the two groups were statistically significant (Z= -2.085, P= 0.037; Z= -2.615, P= 0.009). Logistic regression analysis showed that PNI was an independent risk factor for neutropenia after chemotherapy (HR=0.803, 95％CI 0.646-0.998, P= 0.048). Conclusion PNI has a certain role in predicting neutropenia after adjuvant chemotherapy in patients with colorectal cancer.