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 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 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.

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.

OBJECTIVETo examine the effect of ONO-AE3-208, an EP4 antagonist, on the formation of bone metastasis from prostate cancer in mice.

METHODSThirty-four 6-week old nude mice were divided into an experimental and a control group of equal number to be treated by intraperitoneal injection of ONO-AE3-208 and double distilled water, respectively. Then PC3/LUC cells were constructed by stably transfecting luciferin to prostate cancer PC3 cells and inoculated into the left ventricle of the mice to establish an animal model of systemic bone metastasis. The time of metastasis formation, photon tumor burdens, and changes of the survival curves after modeling were compared between the two groups of mice.

RESULTSAt 30 days after modeling, bioluminescence imaging analysis showed that the photon tumor burdens were significantly increased in a time-dependent manner in the control group in comparison with those in the experimental group (P < 0.01). The rate of metastasis formation was significantly higher in the former than in the latter (93.3% vs 33.3%, P < 0.001). The median time of metastasis formation was 29 d (95% CI 26.547 - 35.262) in the experimental animals as compared with 21 d (95% CI 17.213 -24.787) in the controls (P < 0.001).

CONCLUSIONEP4 antagonist ONO-AE3-208 can inhibit the formation of bone metastasis from prostate cancer in mice.

Animals ; Bone Neoplasms ; prevention & control ; secondary ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Male ; Mice ; Mice, Nude ; Naphthalenes ; pharmacology ; Neoplasms, Experimental ; prevention & control ; Phenylbutyrates ; pharmacology ; Prostatic Neoplasms ; pathology

Objective To evaluate the safety of surgical procedures for patients after heart valve prosthesis implantation.Methods Clinical data of 12 cases with heart valve prosthesis implantation undergone other surgical treatment from November 1996 to December 2005 were retrospectively analyzed.All the cases had routine oral warfarin with prothrombin time (PT) of 20.0—28.3 s averaged 23.5 s, international normalized ratio (INR) for prothrombin of 1.79—2.23 averaged 1.95 and heart functional class Ⅰ—Ⅲ.Among them,appendectomy was performed in three cases with acute appendicitis,reposition and repair in one with inguinal hernia,radical gastrectomy in two with gastric carcinoma,left hemicolectomy in one,cholecystectomy in three,left femoral head replacement in one,and bilateral high ligation and ablation of great saphenous vein in one.Elective surgical operation was performed in seven cases,and emergency operation in five.In those with elective surgery,warfarin was stopped 2—3 days before operation,while 5—10 mg vitamin K_1 was injected intramuscularly 6—8 hours before emergency surgery with preoperative median PT of 15.1 and 15.3 s and median INR of 1.24 and 1.30,respectively.In operation,5—10 mg vitamin K_1 were injected intravenously into the patients by drip depending on their bleeding on the surface of wound.ECG,blood pressure,hemoglobin and oxygen saturation were routinely monitored for all the cases intraoperatively and postoperatively.For the cases with heart function above class Ⅱ,fluid infusion was adjusted based on intubated central venous pressure,and for those with general anesthesia,analyses of blood gases and electrolyte were monitored routinely in operation.Results OPeration time averaged 20—160 rain in all the 12 patients,with blood loss 5—280 ml in average and without complications of massive hemorrhage,thrombosis and heart failure.Conclusions Surgical operation was safe for patients with heart valve prosthesis implantation,if preoperative PT and INR were adjusted to about 15 s and 1.30,respectively by cessation of warfarin or application of vitamin K_1,combined with careful manipulation and strengthened perioperative management.

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.

Objective To evaluate the clinical features and the key points in the diagnosis and management of splenic hemangioma.Methods The clinical presentations,laboratory tests,imaging and pathological results,treatment,and prognosis of 21 cases of splenic hemangioma admitted in Peking Union Medical College Hospital from April,1989 to July 2007 were retrospectively analyzed.Results The clinical presentations of splenic hemangiom are not specific which include left upper quadrant mass or discomfort,abdominal pain,etc.The diagnosis of imaging includes Doppler ultrasound,CT,MRI,DSA,etc.Splencetomy is recommended for all splenic hemangioma with severe symptoms or rupture.Conclusion Asymptomatic patients with small splenic hemangioma(＜4 cm)can be managed conservatively.Symptomatic large hamangioma may need a sp]enectomy.

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.