Objective To establish human neural stem cells(HNSCs) model for further basic research and clinical application. Methods Cells from human fetal cerebral cortex were collected and cultured with serum free midium and then identified for nestin immunocytochemical staning;The cells were induced to differentiate by 5% fetal bovin serum and identified by neurofilament-200(NF-200) and glial fibrillary acidic protein(GFAP) immunocytochemical staning. Results The harvested cells appeared as clusters in suspension and both NF-200 and GFAP positive cells were observed after induction.After 12 generations of culture,these cells retained the main characteristics of NSCs.Conclusions The HNSCs were harvested from human fetal cerebral cortex and this HNSCs model can be used for futher basic research and clinical applications.;

Objective To study the changes of mesenteric arterioles on the contractile response to norepinephrine in patients with cirrhosis and portal hypertension (PHT) caused by hepatitis B after treatment by high-dose vitamin C.Methods Eleven PHT patients and 6 non-PHT volunteer were those undergoing their respective surgeriesat in our hospital from January 2010 to June 2011.The PHT patients were randomly assigned to vitamin C treatment group and placebo treatment group.After admission and before induction of anesthesia,blood taken to mesure plasma concentration of malonaldehyde (MDA).Intraoperative portal pressure was measured.We took the third level intestinal mesentery vessels with their mesentery next to the jejunum and studied the contractile response to norepinephrine of the intestinal mesenteric arterioles.We also measured RhoA/ROCK pathway associated proteins' expression and activity changes by Western-blot method.Results The average value of the plasma MDA of PHT patients with cirrhosis was much higher than that of non-PHT patients.After treatment by vitamin C,the value decreased.The dose-response curve of isolated mesenteric arterioles to norepinephrine from PHT patients with cirrhosis shifted to the right,and the EC50 value was higher than that of non-PHT patients.But this phenomenon was reversed after the patients treated with vitamin C.The protein expression of RhoA in mesenteric arteries did not change significantly among the three groups.But the protein expression and activity of ROCK-1 was significantly reduced in PHT patients with cirrhosis.After treatment with vitamin C,the expression and activity of ROCK-1 in mesenteric arteries of cirrhosis was significantly increased.Conclusion The PHT patients with cirrhosis are in the situation of oxidative stress.The high-dose vitamin C treatment can reduce oxidative stress level and improve contractile hyporeactivity of mesenteric arterioles of PHT patients with cirrhosis to norepinephrine.The protein expression and activity increasement of ROCK in RhoA/ ROCK pathway is involved in thisphenomenon.

Long-term treatment with an agonist of peroxisome proliferator-activated receptor (PPAR)-γ is associated with bone fractures in the clinical practice. However, the mechanisms underlying the fractures are not fully understood. This study was aimed to examine the effect of rosiglitazone (an agonist of PPAR-γ) of different doses on the proliferation, differentiation, and transforming growth factor beta 1 (TGF-β1)-induced expression of connective tissue growth factor (CTGF) in primary rat osteoblasts in vitro. Osteoblasts were isolated from newly born SD rats and treated with different doses of rosiglitazone (0-20 μmol/L). The proliferation and differentiation of osteoblasts were measured by MTT assay and NPP assay, respectively. The expression of CTGF was determined by RT-PCR and Western blotting. The results showed that most isolated osteoblasts displayed strong alkaline phosphatase (ALP) activity and treatment with different doses of rosiglitazone did not affect their proliferation, but significantly inhibited the differentiation of osteoblasts in a dose-dependent manner. Moreover, treatment with different doses of rosiglitazone significantly reduced the TGF-β1-induced CTGF mRNA transcription and protein expression in a dose-dependent manner in rat osteoblasts. It was concluded that the activation of PPAR-γ may inhibit the differentiation of osteoblasts by reducing the TGF-β1-induced CTGF expression in vitro.

BACKGROUND: According to up-to-date development, the best timing to repair cleft palateis at 9-12 months old after birth when the articulation does not begin to develop.However, there are many over-eight-year-old children who still suffer from unrepaired cleft palate. Therefore, it is veryimportant to adopt proper surgery for these patients to improve palate form and articulatory function. OBJECTIVE: To study the characteristics of the cleft palate in patients of over 8 years old and the effects of palate repair and posterior pharyngeal flap transplantation (PPFT) on articulation. DESIGN:A controlled study with children patients as subjects. SETTING: Department of Maxillofacial Surgery, Medical College of Stomatology, Fourth Military Medical University of Chinese PLA PARTICIPANTS: Twenty-four children patients of cleft palate who were treated in the Department of Maxillofacial Surgery ,Medical College of Stomatology, Fourth Military Medical University of Chinese PLA from January 2000 through December 2003 were enrolled in the study. Fourteen were male and ten female. Their ages varied from 8 to 20 years old (mean 13.8 years). Three cases were bilateral complete cleft palate and twelve cases were unilateral complete cleft palate. Nine were incomplete cleft palate. METHODS :Two flap palatoplasty and upper pedicle PPFT were conducted in all the patients. In thirty days after surgical treatment, the patients were asked to repeat the words of a doctor in the phonetic lab with his or her lips 10 cm away from microphone. The words of the patients were recorded and evaluated subjectively by 5 doctors specialized in pathologic phones. MAIN OUTCOME MEASURES :The palatopharynx was observed with nasopharyngoscope before and after operation. And the improvement of nasal gas leakage and enhanced rhinophonia were also evaluated. RESULTS: The repaired cleft palate was healed in the first intention in all the patients. The soft palate retreated satisfactorily and the palatopharynx was apparently closed, which created good preconditions for articulation. The patients' articulation was also improved in different degrees. All the 24 patients had the fourth grade of enhanced rhinophonia and nasal gas leakage before repairing. After palate repair and PPFT, the enhanced rhinophonia and nasal gas leakage of grade 1 was in 3 patients, grade 2 in seven, grade 3 in six and grade 4 in eight patients. CONCLUSION: The combination of palate repair and pharyngoplasty is able to improve the form of soft palate and the articulation of over-eightyear-old patients with cleft palate.

Objective To analyze the impact of medical insurance reimbursement mechanism on the income of urban community health centers run by various entities. Methods The data of the baseline survey of community health centers in 28 cities made in 2007 were called into play. These data were used to compare the percentage of the annual medical insurance income from outpatient clinics of three types of community health centers in their gross annual income and their annual outpatient clinic income. Results For 929 community health centers with income from medical insurance reimbursement, the average percentage of their annual income from outpatient clinic accounts for 24% of their gross annual income and 26. 09% of their annual outpatient income;In respect of the reimbursement rates of medical insurance at outpatient clinics, the highest come from those run by enterprises, secondly those by the government,and the least those run by social organizations or individuals. Conclusion The mechanism of medical insurance reimbursement has already affected the income of the third type of community health centers. It is recommended to raise the coverage rate of medical insurance of such type of centers and their reimbursement rates of medical insurance. This may further consolidate the policies of medical insurance reimbursement.

Objective To study the role of cell density in the tyrosol production and morphology for Candida albicans biofilms. Methods C. albicans SC5314 and clinical isolates were propagated in yeast peptone dextrose (YPD) medium. Cells were collected by centrifugation and washed twice in sterile phosphate-buffered saline (PBS) before this study, then resuspended in RPMI 1640 supplemented with L-glutamine and adjusted to a desired concentration of 5 × 10~6 cells/ml, 5×10~5 cells/ml, 5 × 10~4 cells/ml, 5 × 10~3 cells/ml after counting with a hematocytometer. Standardized C. albicans cells were prepared as above description and 2000 μl of this standardized cell suspension was dispensed into the wells, then C. albicans biofilms were formed on the bottom of the polystyrene wells. In this study, tyrosol synthesized by SC5314 and clinical isolates of C. albicans biofilm was quantified by high performance liquid chromatography (HPLC). The effects of tyrosol on morphology of C. albicans biofilms were investigated by scanning electron microscopy (SEM). Results Tyrosol production of C. albicans biofilms was affected by cell densities. At lower inoculation size(5 μ 10~3 cells/ml), there was too less tyrosol production to be detected at the early stage of the biofilms formation. At higher inoculation size (5 μ10~6 cells/ml), tyrosol can be detectable at the early stage or at the mature stage of biofilms formation. There was a sharp increase in tyrosol concentration at 24 h, while there was a decrease in tyrosol concentration after that time from the strains when the strains were at an inoculation size of 5 × 10~6 cells/ml and 5 × 10~5 cells/ml. Cell densities affected the morphology formation of the C. albicans biofilms. At the early stage of the biofilms formation, C. albicans grew less germ tube at lower cell densities than that at the higher cell densities. With the mature of the biofilms, C. albicarts grew more hyphae at higher cell densities than that at the lower cell densities. All these above showed that cell densities played an important role in the propagation for the C. albi-cans in the biofilm formation. Conclusion Cell density play an important role in the formation of the C. albi-cans biofilms and the production of the tyrosol.

Objective To evaluate the effect of ultra-low dose naloxone on postoperative hyperalgesia caused by large-dose remifentanil.Methods Forty ASA Ⅰ-Ⅲ adult patients,scheduled for gastrointestinal surgery,were randomly assigned into 2 groups (n =20 each):large dose remifentail group (group R) and ultra-low dose naloxone group (group N).Anesthesia was induced with iv injection of remifentanil,propofol and cisatracurium and maintained with inhalation of sevoflurane and infusion of remifentanil.The patients were tracheal intubated and mechanically ventilated.In group R,remifentanil was infused at a rate of 0.25 μg· kg-1 · min-1 starting from the beginning of skin incision.The infusion rate was adjusted according to hemodynamics during operation and subsequently increased/decreased by 0.05 μg· kg-1· min-1 each time.In group N,naloxone was infused at 0.1 μg·kg-1· h-1 while infusing remifentanil,naloxone infusion was stopped at the beginning of peritoneum closure and the other treatments were similar to those previously described in group R.All patients were sent to post-anesthesia care unit after surgery and stayed there for 90 min.Morphine was given when need.The patient-controlled intravenous analgesia was used for postoperative analgesia after leaving post-anesthesia care unit.The first pain time was calculated.The morphine consumption and complications such as nausea,vomiting and pruritus were recorded at 15,30,60 and 90 min and 2,6,24,48 and 72 h after surgery.Results Compared with group R,the morphine consumption was significantly reduced at each time point after surgery,the first pain time was prolonged,and incidence of nausea was decreased (P ＜ 0.05),while no significant change was found in the incidence of vomiting and prutirus in group N (P ＞ 0.05).Conclusion Infusing ultra-low dose naloxone (0.1μg· kg-1 ·h-1) during operation can attenuate postoperative hyperalgesia caused by large-dose remifentanil in patients.

Objective To investigate the compliance with endocrine therapy(ET) and assess the factors associated with treatment accuracy for breast cancer.Methods 379 patients with hormone receptor-positive breast cancer undergoing complete treatment from Jun.2006 to Jun.2008 in Jiangsu Cancer Hospital were followed up.Factors related to compliance were analyzed.Results Among the 322 (85.0％) patients successfully interviewed,15 (4.7％) patients did not receive ET,43 (13.4％) patients stopped taking drugs after discharge,14 (4.3％)patients had intermittent ET,and the rest 250 patients obeyed 5-year oral ET regularly.The treatment accuracy was 77.6％ (250/332).We found that majority of withdraw occurred within 2 months and within 2 years,accounting for 39.5％ and 48.8％,respectively.Job and education status were relevant to these patients.Moreover,too much concern of adverse drug effects and difficulty of long-term medication were the main reasons to noncompliance.Conclusions ET is effective in hormone receptor-positive breast cancer patients and has been used as a conventional therapy.However,due to the long therapeutic period and lack of medical supervision after discharge,its treatment accuracy and compliances are becoming low,resulting in decreased efficiency.It is therefore necessary to investigate such patients' management to improve the compliance and treatment accuracy of ET.

Objective To review our clinical experience with primary surgical repair of tetralogy of Fallot in neonates and premature infants and to discuss the timing of repair and major factors in treating this patients perioperatively.Methods From January 2012 to September 2015,a series of 19 consecutive neonates and premature infants(M/F =12/7) with tetralogy of Fallot were admitted in our center to receive surgical treatment,with a mean age of(17.3 ± 5.5) days(12-28 days) and a mean body weight of(2.9 ±0.7) kg(2.1-4.3 kg).All the 19 cases were symptomatic with cyanosis,saturation on room air 0.79 ± 0.12 (0.48-0.92),and shortness of breath.Before operation,2 cases were receiving an infusion of prostaglandin E1,5 were mechanically ventilated.7 were more than moderate anemia with hemoglobin of 55-87 g/L.All the patients received echocardiography,ECG and chest X-ray.The McGoon ratio and Nakata index were 1.09 ± 0.30 (0.8 to 1.6) and (135.5 ± 54.2) mm2/m2 (63-212 mm2/m2) respectively.18 cases received one-stage surgical repair and 1 premature infant under two-stage operation with the VSD closure after right ventricular outflow tract(RVOT) transannular patch augmentation.All the VSDs in the 18 cases were closed with continuous suture and RVOT were enlarged with autologous pericardium patch transannularly or not.Balanced and modified ultrafiltration were applied in all the patients.Patients were strictly followed up with a standard protocol focusing on right ventricular function and arrythmia.Several characteristics(e.g.time of operating,mechanical ventilation and ICU stay,complications,hospital stay time,cost of hospitalization) were compared between this group of patients and other TOF patients during the same period in our center.Results All the one-stage operations were successful.There was no mortality and major complication.Mean CPB and aortic clamping time were(111.5 ± 31.6)min (76-153min) and (73.3 ± 11.6) min (64-89 min) respectively.10 VSDs were closed with transventricular approach,6 witht transatrial appraoch and 2 with transatrial-ventricular approach.12 cases (66.7 ％) had a transannular RVOT patch,4 (22.2 ％) with single RVOT patch and 2 (11.1％) with transannular RVOT and left pulmonary artery patch.Atrial communication were left open in 15 cases (83.3 ％).The time of mechincal ventilation and ICU stay were (123.7 ± 59.5) h (39-239 h) and (10.1 ± 3.2) days (5-19 days) respectively.All the patients were followed up for (31.8 ± 15.7) months (9-57 months).There was no mortality and major complication.ECG showed that there was no severe arrythmia except for 3 complete right branh bundle block.The latest echocardiography results showed that right heart function was normal in all the cases and RVOT grandients was less than 30mmHg except one with 35 mmHg.There were 2 cases with residual shunt less than 2 mm and 8 cases with pulmonary valvular regurgitation less than moderate degree.Compared with other TOF cases during the same period,there was no difference according to the data mentioned above except with more time of mechnical ventilation and ICU stay and more cost of hospitalization.Conclusion Primary repair of TOF can be performed safely in symptomatic neonates and premature infants,regardless of age and body weight,with favorable early and mid-term results.Excellent teamwork and accurate prenatal and postnatal diagnosis were the two major factors in yeilding good results in these patients.

Objective To study the expression of PI3K and Akt mRNA in acute leukemia(AL) and chronic myelocytic leukemia (CML) and try to find out a relationship between the prognostic significance of acute leukemia and the expression level of PI3K and Akt. Methods The samples were collected from 63 ALpatients, 7 relapsed AL patients, 14 CML patients and 10 AL patients in complete remission(CR) patients, 11samples of normal controls(NC). The expression of PI3K, Akt mRNA were measured by semi-quantity reverse transcription polymerase chain reaction(RT-PCR). Results The expression of PI3K mRNA in AL and relapse group were significantly higher than that in normal control. In CML, the P13K mRNA expression level was higher than that in NC and CR. The similar results of the expression of Akt mRNA were observed in CML, NC and CR. At the same time, the expression level of Akt mRNA in relapse group was significantly higher than that in NC. The complete remission rate in PI3K (+) was lower than that in PI3K (-). Similar result was obtained in Akt (+) and Akt (-). Conclusion The PI3K/Akt pathway may contribute to the occurrence of leukemia. The patients with positive expression of the members of this pathway had a lower remission rate.