Objective To investigate the anti-fatigue and anti-hypoxia effect of fermented Paecilomyces hepiali myceli-um (PHM) and to explore its mechanism.Methods Seventy-two naive Kunming male mice were randomly divided into Home, vehicle, modafinil( Mode) , low-dose PHM, moderate-dose PHM, and high-dose PHM groups.All mice were ad-ministrated with the drug or vehicle twice a day during a 14-day period.Except for the Home group, each groups was forced into the wheel fatigue device to receive a climb-run training at the time points of 40 min, 8 h, 24 h and 48 h, and to re-ceive an exhaustive training at the time point of 72 h after the last administration.The number of electric shocks was recor-ded during each training and the levels of muscle glycogen, liver glycogen, serum urea nitrogen, serum lactic acid, serum creatine kinase and blood ATP were detected after the exhaustive training using commercially available kits.In addition, 96 naive Kunming male mice were randomly and equally divided into the vehicle, low-dose, moderate-dose and high-dose PHM groups.Twelve mice in each group were chosen for the normal pressure anti-hypoxia experiment and the other 12 mice were used for the NaNO3 poisoning experiment.Results Compared with the vehicle groups, the PHM group showed a sig-nificant decrease in the number of electric shocks, improvement in biochemical parameters associated with fatigue, and an increased survival time in the anti-hypoxia and the NaNO3 toxicity tests.Conclusion PHM is potentially an effective alter-native for wild Cordyceps in the treatment of fatigue and hypoxia.

Objective To investigate the effect of three-dimensional gelatin-chondroitin sulfatehyaluronic acid (Gel-C6S-HA) composite scaffold seeded with genetically modified hair follicle stem cells (HFSCs) on tissue engineering skin angiogenesis.Methods Three-dimensional scaffolds composed of Gel-C6S-HA were fabricated by freeze-lyophilizing.VEGF165 modified HFSCs were seeded on those scaffolds and cellular morphology and adhesion were observed using scanning electron microscope.Eighteen rats were subjected to the full-thickness skin defects with dimensions of 1.2 cm × 1.2 cm on the bilateral sides of the back and covered with VEGF165 transduced HFSCs/Gel-C6S-HA scaffold (Group A),empty-vector transduced HFSCs/Gel-C6S-HA scaffold (Group B),Gel-C6S-HA scaffold (Group C),and vaseline gauze (Group D) respectively according to the random number table.At days 7,14,and 21 after surgery,wound healing was observed,HE staining and immunohistochemistry of CD31 and alpha smooth muscle actin (α-SMA) were performed,and microvessel density (MVD) was used to measure new blood vessel growth.Results Electron microscopy exhibited three-dimensional spongy structure of the scaffold with round or polygon apertures connecting mutually and the scaffold pore size of (133.2 ± 43.4) μm.Cells seeded on the scaffolds spread thoroughly out and anchored firmly after being cultured for 7 days.There were no obvious inflammation reactions on the wounds for all groups at day 7 after operation.Wound healing and scaffold degradation were faster in Group A than in other groups at days 14 and 21 after operation.Histological and immunological detections showed microvasculariztion of the scaffold in Groups A and B with fluffy three-dimensional structure and evenly distributed cells,but scaffolds remained sharp at the edge and there were small cells aggregating at subcutaneous tissue junction area in Group C.At 14 and 21 days after operation,new blood vessels were large and rich in Group A; scaffolds in Groups A and group B were filled with cells,partial of which gathered in the epidermal layer and the scaffold materials were assimilated differentially,whereas a few subcutaneous tissue cells migrated to scaffolds in Group C.MVD was significantly higher in Group A than in Groups B and C at each time point (P ＜ 0.05).Conclusion VEGF165 modified HFSCs compounded with Gel-C6S-HA composite scaffold can facilitate the growth of blood vessels and promote the angiogenesis in wound healing and hence is a promising skin substitute in clinical applications.

Objective To study the clinical features of gallbladder small cell carcinoma (GSCC),to improve the diagnosis and treatment of GSCC.Methods We retrospectively analyzed the clinical data of GSCC patients at our hospital from January 2000 to January 2012,and made a collective review of the literature.Results In this series,there were four female cases,one male case,the age at the first diagnosis was between 42-67,with the median age of 57.The main complain was pain and dis-comfort on the up and right abdomen.Tumor located in the bottom of gallbladder in 3 cases,and in the body in 2.Cholelithiasis was complicated in 4 cases.2 patients received radical resection of GSCC,followed by adjuvant chemotherapy of VP-16 and cisplatin,radioactive therapy in one.Postoperatively,these two were followed up for 45 and 32 mons with tumor free survival.3 cases received palliative resection,followed by adjuvant chemoradioactive therapy or intervention treatment,these three were followed up to 8,11,30 months respectively to their death for tumor recurrence.Conclusions GSCC is a rare disease,the initial symptoms are not often specific and easily misdiagnosed.The prognosis of GSCC is poor.

Objective To summarize and analyze the clinical feature,therapeutic mnethods and prognosis of colorectal small cell carcinoma.Methods From January 2000 to January 2012,15 patients of colorectal small cell carcinoma were analyzed retrospectively.Results There were 12 male cases,3 females.The age at diagnosis was between 39-71 years,with median age of 60.SCC located in the rectum in 12 cases,in the colon in 3 cases.The time from the onset of symptoms to final diagnosis was from 1 to 12 months.The diameter of tumors varied from 2.5 to 8.0 cm.13 cases received up-front surgery,including radical tumor resection in 6 cases,palliative resection in 7 cases,and neoadjuvant-chemotherapy followed by palliative resection in one case.The initial Ⅰ,Ⅲ B,Ⅳ B stage were 1 case,6 cases and 8 cases,respectively.The overall median survival time is 11 months,1,2 year's survival rate is 40.0％ and 20.0％,respectively.Conclusions Colorectal SCC is less common and the prognosis is poor.Multimodality management,with radical surgical resection of the primary lesion followed by standard adjuvantchemotherapy,affords good local disease control and a fair survival.

Objective To study the effects of electrical stimulation of efferent vagus nerve on the endotoxic shock in rabbits. Method Sixteen Newzealand rabbits were randomly divided into 2 groups, namely group S as the stimulation group and group C as the control group). Rabbits were subjected to bilateral cervical vagotomy and had challenge with intravenous injection of lipopolysaccharide (LPS) (E. COLI O111: B4 , DIFCO, USA) in a dose of 600 μg/kg. The distal end of the left vagus nerve trunk was connected to an electric stimulator with bipolar electrode and controlled by an acquisition system. Stimuli with stable voltage (10 V,5 Hz,5 ms) were applied twice to the nerve for 10 minutes just before and after the administration of LPS in group S. At the time before and after the infusion of LPS 30 min,60 min, 120 min, 180 min,240 min and 300 min respectively, the heart rate(HR)and the mean arterial blood pressure (MABP) in each animal were recorded, and blood samples were taken for measuring serum tumor necrosis factor-αa(TNF-α) and interleukin-10 (IL-10). Results Compared with group C,the electrical stimulation of efferent vagus nerve could significantly attenuated the LPS-induced hypotension and de-creased the contents of TNF-α[(38.12±7.85) pg/mL vs. (55.12±7.89) pg/mL, P <0.01], but increased the contents of IL-10[(55.12±9.37)pg/mL vs. (40.15±5.44) pg/mL, P <0.01]afar LPS challenge. Conclusions The stimulation of the efferent vagus nerve can down-regulate systemic TNF-α, production and attenu-ate the development of shock after LPS challenge.

OBJECTlVE To investigate the effect of Guiyuan tablets on the analgesic effect of morphine and morphine-induced tolerance and hyperalgesia. METHODS ① The model of morphine-induced acute tolerance Mice were ig treated with Guiyuan tablets 200, 400 and 800 mg·kg-1 and 15 min later were sc treated with morphine 10 mg·kg-1 every hour for consecutive 9 h. At 24 and 48 h, they were sc treated with morphine 10 mg·kg-1 alone, respectively.②The model of morphine-induced chronic tolerance Mice were ig treated with Guiyuan tablets 200, 400 and 800 mg·kg-1 and 15 min later were sc treated with morphine 10 mg·kg-1 every day for consecutive 8 d. On d 9, the mice were sc treated with morphine 10 mg·kg-1 alone. ③ The model of morphine-induced established tolerance. Mice were sc treated with morphine 10 mg·kg-1 every day for consecutive 8 d. On d 1, d 4 or d 7, the mice began to be ig co-administered with Guiyuan 200 mg·kg-1 . On d 9, they were sc treated with morphine 10 mg·kg-1 alone. The hot-plate test was used to detect the values of the baseline latency ( T0 ) and the post-treatment latency ( T1 ) before the percentage of maximal possible analgesic effect (%MPAE) was calculated. Spec-trophotometry was used to detect the nitric oxide synthase ( NOS) activity and the nitric oxide ( NO) con-tent in the tissue of the spinal cord. RESULTS The ED50 of the analgesic effect of Guiyuan tablets was 523.5 mg·kg-1 in the hot-plate test. Guiyuan tablets 200 and 400 mg·kg-1 prolonged the duration of mor-phine anti-nociception and deceased its ED50 from 4.67 to 3.14 and 0.65 mg·kg-1, respectively. ln the models of both acute and chronic tolerance, Guiyuan tablets prevented the decrease of the%MPAE and the baseline latencies ( P<0. 05 ) . ln the model of morphine-induced established tolerance, Guiyuan tablets rapidly reversed the decrease of%MPAE( P<0.05) , and this compound preparation which began to be co-administered with morphine from d 1 could significantly inhibit the increase of the NOS activity and NO content induced by morphine in the spinal cord ( P<0.01) . CONCLUSlON Guiyuan tablets are capable of enhancing the analgesic effect of morphine, prolonging the duration of morphine anti-nocicep-tion, preventing the development of morphine-induced tolerance and hyperalgesia, and might have neuroprotective effect.

ObjectiveTo evaluate the safety and effects of pseudomonas aerug PA-MSHA vaccine on cytoimmunity in advanced gastric cancer patients. MethodsSeventy two patients with advanced gastric cancer were randomly divided into experimental group and control group. Tumor bed was treated intraoperatively by pseudomonas vaccine injection in experimental group.The venous bloods were sampled prior to surgery (preoperative) and on days 10,20,and 30 postoperatively.The total lymphocyte count and lymphocyte subpopulations were detected while the postoperative complications and adverse drug reaction were observed,prognosis was evaluated. ResultsThe total lymphocyte count(P =0.042) and the counts ofCD3+ lymphocyte(P =0.027) and NK cell increased (P =0.012) obviously in experimental group.CD8+ lymphocyte decreased(P =0.037),but the counts of CI4+ lymphocyte and CD4+/CD8+ were not significantly different.Complications were not significantly different in the two groups.One year survival rate was longer ( 94.9％ ) in experimental group than that in the control group ( 83.3％ ) ( P =0.022 ).ConclusionsPeritoneal cavity administration with pseudomonas aerug vaccine was safe,and effectively helps regulate cytoimmunity in postoperative patients of advanced gastric carcinoma.

Objective At present, there is no study on effect of levetiracetam(LEV) on the gray matter structure remodeling in benign epilepsy children with central temporal spikes(BECTS).The purpose of this study was to study the influence of LEV on the gray matter structure in BECTS and to evaluate the mechanism of LEV on the brain structure of BECTS through using voxel-based MRI morphological(VBM) methods.Methods From January 2014 to September 2016, twenty-four BECTS treated with LEV(LEV group), twenty-four drug-na?ve BECTS(untreated group) and twenty-four normal children(normal group) consulted in department of Neurology, Nanjing Children′s Hospital and the Nanjing Military Region, Nanjing General Hospital were continuously included to receive three-dimensional T1-weighted imaging with 3T MRI and the gray matter volume was calculated by VBM.We compared the difference of grey matter volumes of the three groups and analyzed their correlation with epilepsy duration, age of onset and medication time and other clinical index.Results Compared with the normal group, the grey matter volume of bilateral thalamus were decreased, and the volume of bilateral Rolandic areas, anterior insula/frontal operculum/frontal triangle, left supplementary motor area, paracentral lobule, precentral gyrus, superior frontal gyrus and right middle frontal gyrus were increased in the untreated group, but the grey matter volume of the bilateral Rolandic areas, frontal operculum and left supplementary motor area were decreased in the LEV group.Compared with the untreated group, the grey matter volume of bilateral supplementary motor, left paracentral lobule, precentral gyrus, bilateral anterior insula/frontal operculum/frontal triangle, left superior frontal gyrus and right middle frontal gyrus in the LEV group were decreased.The grey matter volume of left anterior insula/frontal operculum areas was negatively correlated with the medication time in LEV group(r=-0.527, P<0.01).Conclusion T The mainly representations of BECTS are thalamic gray matter damage and epileptic-related cortical area irritation structural abnormalities, but the LEV could reshape the epilepsy-related cortical area and the gray matter in the brain area associated with clinical symptoms.

AIM: To investigate the changes in nNOS and iNOS expression of hippocampal CA3 pyramidal neurons and NO - 2/NO - 3 level of hippocampal homogenate of rats induced by stress, and to explore the effect of phenytoin on them. METHODS: Rats were subjected to forced-swimming stress, phenytoin was administered(ip) at 30 min before stress. Using the immunohistochemistry and the computerized image technique, the expression levels of nNOS and iNOS of rat hippocampal CA3 pyramidal neurons were assayed quantitatively, and the NO - 2/NO - 3 level of hippocampal homogenate was also measured using nitric acid deoxidize enzyme method. RESULTS: The nNOS average grey degree of hippocampal CA3 pyramidal neurons was significantly lower in stress group (155 42?3 77)than that in control group(164 54?4 62)and in stress plus phenytoin group(164 27?2 55)( P

Pseudomonas sp. M18, one of plant-growth-promoting rhizobacteria, can produce secondary metabolites including phenazine-1-carboxylic acid (PCA) and pyoluteorin (Plt). PA2572 gene coding protein is a probable two-component response regulator in Pseudomonas according to homologous speculations. In order to investigate its genetic function, PA2572 homologous gene, ppbR, was amplified from M18 genome, inactivated by inserting a Gm cassette. The resulting reconstruct was introduced into the M18 genome using homologous recombination technique, so as to obtain the null mutant M18P. The results showed that the M18P has less flagellar swimming and swarming motility, and yielded fewer PCA. The production of PCA was only 50% of the wild type. However, there was no remarkable difference between mutant and wild type in producing pyoluteorin in KMB medium.