Abstract:Objective To investigate clinical features of 237 gastric cancer patients from plateau region in Tibet. Methods Clinical information of 237 patients with gastric cancer admitted to hospitals in the Tibet region was retrospectively analyzed. Clinical features including sex, age, clinical, endoscopic and histopathologic compositions were statistically compared. Results In all patients, the sex ratio of male to female was 3.2:1; 51.5% were aged from 40 to 59 years; 60.8% were peasants or cadres; 68.4% had tumors in gastric antrum; 82.3% had infiltrated type of gastric cancer; 81.0% presented with lowly-differentiated adenocarcinoma; the positive rate of CEA (carcino-embryonic antigen, as a tumor marker) was 30.0%; 95.2% had stages III to IV cancer evaluated based upon TNM criterion (tumor, node, metastasis). Conclusion In the Tibetan plateau, gastric cancer was commonly seen in gastric antrum of middle-aged or senior men. The severity of carcinoma was high. Effective and comprehensive measures should be actively adopted to prevent and treat gastric cancer in the plateau region according to local clinical features.

Abstract:Objective To construct a small interfering RNA (siRNA) vector targeting ΔNp63α and investigate ΔNp63α gene interference on the proliferation and apoptosis of human esophageal squamous carcinoma Eca109 cell line. Methods Adeno-associated virus (AAV)-ΔNp63α^shRNA driven by H1 promoter was constructed and was used to infect Eca109 cells. AAV-Null and normal cell lines were utilized in the control group and blank control group, respectively. The influence of siRNA interference of ΔNp63α expression on the growth, proliferation, tumorigenic efficiency and apoptosis of Eca109 cells were analyzed in vitro and in vivo. Results Compared with the two control groups, the specific siRNA targeting ΔNp63α gene significantly down-regulated the expression of ΔNp63α protein levels in Eca109 cells (all P<0.05).The growth of Eca109 cells infected with AAV-ΔNp63α^shRNA was significantly lower than those in the two control groups (all P<0.05). Cell cycle analysis showed the proliferation index (PI) of AAV-ΔNp63α^shRNA infected cell line was significantly lower compared with the two control groups (all P<0.01). In vivo experiment exhibited that AAV-ΔNp63α^shRNA infected cells resulted in a lower tumor weight in nude mice compared with the cells in the two control groups (all P<0.05). In addition, the apoptosis index (AI) of AAV-ΔNp63α^shRNA infected cells were significantly higher than those of the other cell lines (P<0.05). Conclusion AAV-mediated expression of siRNA can significantly reduce ΔNp63α expression in Eca109 cells, slowing down the proliferation, promoting the apoptosis, and subsequently inhibiting the growth of tumor.

Abstract:Objective To investigate the chemical components of gorgonian Menella kanisa collected from the South China Sea. Methods The Et₂O extract of gorgonian Menella kanisa was purified by repeated column chromatography on silica gel, Sephadex LH-20, and reversed-phase high-performance liquid chromatography (RP-HPLC). The structures of the obtained compounds were determined on the basis of spectroscopic analysis and compared with the reported data. Results and Conclusion Five 5α,8α-epidioxy sterols were isolated from gorgonian Menella kanisa collected from the South China Sea, and their structures were determined as: 5α,8α-epidioxy-cholest-6-en-3β-ol (1), (22E)-5α,8α-epidioxy-cholesta-6,22-dien-3β-ol (2), (22E,24R)-5α,8α-epidioxy-24-methyl-cholesta-6,22-dien-3β-ol (3), 5α,8α-epidioxy-24-methyl-cholesta-6,24(28)-dien-3β-ol (4), (24E)-5α,8α-epidioxy-24-ethyl-cholesta-6,24(28)-dien-3β-ol (5). All the five epidioxy sterols have been separated from the title Menella kanisa for the first time.

Abstract:Objective To examine whether cyclosporine A (CsA) treatment can induce anemia and changes in expression of erythropoietin (EPO) and its receptor (EPOR) in renal tissues of rats with chronic CsA nephrotoxicity. Methods Sprague-Dawley rats were treated daily with subcutaneous injection of vehicle (olive oil, 1 mL/kg, control group) or CsA (15 mg/kg, CsA nephrotoxicity model group) for 4 weeks. The renal function and hemoglobin (Hb) and hematocrit (Hct) levels were analyzed by Automatic Biochemistry Analyzer in the two groups. Masson Trichrome staining was used to examine the extent of tubulointerstitial fibrosis. Immunohistochemistry and Western blotting analysis were used to examine the expression of EPO and EPOR protein. And cell apoptosis was observed by TUNEL assay and electron microscope. Results Compared with the control group, CsA nephrotoxicity group showed renal insufficiency, tubulointerstitial fibrosis, and significantly more apoptotic cells (P<0.01). Meanwhile, CsA nephrotoxicity group also had anemia, manifested by significantly decreased Hb and Hct levels (P<0.01). Immunohistochemistry and Western blotting analysis showed that CsA treatment significantly decreased EPO expression and significantly increased EPOR expression compared with the control group (P<0.01). Furthermore, we also found that EPO protein expression was negatively associated with tubulointerstitial fibrosis (r=-0.729, P<0.001) and the number of TUNEL-positive cells (r=-0.841, P<0.001). Conclusion Renal EPO expression is decreased in rats with chronic CsA nephrotoxicity, which resulting in anemia; and tubular epithelial cell apoptosis induced by CsA is related to decrease of EPO protein.

Abstract:Objective To explore the effects of vitamin A combined with other micronutrients on the nutritional status in children. Methods According to the method of stratified random cluster sampling, three kindergartens were randomly selected out of 7 in the suburbs of Chongqing. A total of 350 preschool children who met the eligibility criteria, aged between 3-6 years old in the selected kindergartens, were randomly assigned to 3 treatment groups: the vitamin A group (A group), the vitamin A plus Zn group (AZ group), and the vitamin A supplementation with multiple micronutrients group (AMM group). The supplementation lasted for 6 months in the three groups. The height and weight of the participants were measured and the Z-score value was calculated before and after supplementation to evaluate the incidence of malnutrition. Serum vitamin A, Zn, Fe, Ca and hemoglobin levels were also measured before and after the intervention. A questionnaire survey of the general situation, family status and dietary habit of the children was performed before the supplementation; also the dietary nutrient intake of the children during intervention was investigated by a 24-hour dietary recall method. Results The 24-hour dietary recall results showed that vitamin A, Zn and Ca were inadequate in the diet of the 3-6 years old children in our study. Compared with before supplementation, the serum Zn and Fe levels were significantly different in the three groups 6 months after supplementation (all P<0.01), with the increment of Zn level in AZ group being significantly higher than that in the AMM group (P<0.05). The serum vitamin A level in AZ group and AMM group were significantly increased by (0.05±0.23) μmol/L and (0.09±0.28) μmol/L (P<0.05, P<0.01), respectively; however, the increase was (0.03±0.27) μmol/L in A group (P>0.05). The increment of serum vitamin A in AMM group was significantly greater than those in A and AZ groups (all P<0.05); the increment of hemoglobin level in AMM group was also significantly greater than that in A group (P<0.05).The malnutritions rates were decreased in the A, AZ and AMM groups to different degrees, and the decreases were not significantly different between the three groups (P>0.05). Conclusion The effect of multiple micronutrient supplementation shows no significant difference on nutritional status in 3-6 aged children compared with the two micronutrient supplementations. Micronutrient supplement for children should be based on the main nutritional problems of them, and the best combination of nutrients should be chosen to improve nutrition and health status of children.

Abstract:Objective To evaluate the efficacy and safety of transarterial chemoembolization (TACE) combined with sorafenib in treatment of patients with non-resected middle stage hepatocellular carcinoma (HCC). Methods From July 2009 to July 2011, 38 middle stage HCC patients treated with TACE plus sorafenib were included in the TACE+sorafenib group and the other 38 patients with similar baseline characteristics receiving TACE alone were selected in the TACE group. All the patients belonged to the Barcelona Clinic Liver Cancer (BCLC) stage B. The adverse events of the TACE+sorafenib group and the overall survival of the two groups were analyzed. Results All the patients treated with sorafenib experienced at least one drug-related adverse event during the study, including 7 (18.4%) experienced drug-related grade 3 adverse events, with no grade 4 or higher adverse events occurred. The median overall survival time was 11 months (95% CI: 7.4-14.6 months) for the TACE group and 15 months (95% CI: 8.4-21.6 months) for the TACE+sorafenib group, showing significant difference between the two groups (P=0.019). Conclusion The combination of sorafenib with TACE is well tolerated in non-resected middle stage HCC patients, with no severe adverse events and can effectively improve the overall survival time of patients.

Abstract:Objective To observe the correlation between chronic obstructive pulmonary disease (COPD) assessment test(CAT) score and prognostic factors, so as to investigate the value of CAT score in predicting the prognosis of COPD. Methods A total of 81 patients with newly diagnosed COPD in our hospital during Jul.2011 to Sep.2012, without using inhaled corticosteroid (ICS)/long-acting β₂ agonist (LABA) or long-acting antimuscarinic agent (LAMA), were divided into group A (low risk, less symptoms), B (low risk, more symptoms), C (high risk, less symptoms) and D (high risk, more symptoms) groups according to Global Initiative for Chronic Obstructive Lung Disease (GOLD, 2011 edition), and the patients were given ICS/LABA or ICS/LABA+LAMA treatment for 3 months. The CAT score, age, smoking quantity, pulmonary function indices, body mass index (BMI), 6-min walking distance (6MWD), modified medical British research council (mMRC) dyspnea scale, and the times of acute exacerbation of COPD (AECOPD) in previous one year were collected before and after treatment. The clinical characteristics analysis and correlation analysis were performed. Results The average age of the 81 COPD patients was (66.27±8.52) years, with 88.89% being males and 85.19% having smoking history. The proportions of group A, B, C and D were 8.64%, 30.86%, 4.94% and 55.56% before treatment, respectively. The values of the forced expiratory volume in one second (FEV₁), predicted amount as a percentage of FEV1 (FEV₁%Pred), forced vital capacity (FVC), predicted amount as a percentage of FVC (FVC%Pred), peak expiratory flow (PEF), predicted amount as a percentage of PEF (PEF%Pred), and 6MWD in CAT score ≥10 groups were significantly less than those in CAT score<10 group (P<0.05). The above parameters were not significantly different between patients with CAT score being 10-20, 20-30 and ≥30 groups. mMRC scale and times of AECOPD in CAT score ≥20 groups were significantly higher than those in CAT score<10 group (P<0.05). No significant difference in FEV₁/FVC was found in different CAT score groups. The CAT score was significantly correlated with mMRC scale (pre-treatment r2=0.417, P<0.001; post-treatment r2=0.19, P<0.001), 6MWD (pre-treatment r2=0.320, P<0.001；post-treatment r2=0.19, P<0.001), pre-treatment FEV1 (r2=0.177, P=0.001 5), FEV1%Pred(r2=0.125, P=0.002), PEF(r2=0.164, P=0.002 4), PEF%Pred (r2=0.129, P=0.007 6), FVC (r2=0.098, P=0.021), FVC%Pred (r2=0.094, P=0.024), FEV1/FVC(r2=0.101, P=0.005 7), and AECOPD number (r2=0.059, P=0.028); and not correlated with the quantity of smoking (r2=0.041, P=0.083), BMI (r2=0.00, P=0.89), and post-treatment FEV1(r2=0.01, P=0.22) or FEV1%Pred (r2=0.003, P=0.09).Conclusion COPD is prone to occur in the male smokers, with the highest proportion found in group D. CAT score has a good correlation with pre- and post-treatment mMRC scale and exercise capacity, suggesting it has a potential for predicting prognosis of COPD.

Abstract:Objective To investigate the protective effects of andrographolide on septic mice and to explore the underlying mechanism. Methods Sepsis model was induced by cecal ligation and puncture (CLP) in mice. A total of 64 C57BL/6 mice were evenly randomized into 4 groups: a sham group, a CLP group, a dimethyl sulfoxide (DMSO) group and an andrographolide (AND) group. The mice were injected intraperitoneally with 0.2 mL normal saline, 0.2 mL 5% DMSO or 0.2 mL andrographolide (10 mg/kg) at 1 h, 6 h and 12 h after surgery. The survival rates were assessed 7 days after surgery and the survival curve was plotted. Another 48 C57BL/6 mice were grouped as above. The peripheral blood, peritoneal lavage fluid (PLF), lung and liver tissues were harvested 24 h after operation. Polymorphonuclear leukocyte (PMN) count in PLF was determined. Bacterial loads in the peripheral blood sample and PLF were also determined. The levels of TNF-α, IL-6 and IL-10 in the blood samples were detected by ELISA. The lung and liver tissues were observed and analyzed by hematoxylin-eosin staining. Results Compared with CLP group, AND group had a significantly increased survival rate 7 days after operation (P<0.01), a significantly decreased PMN count in PLF (P<0.01), a significantly enhanced bacterial clearance capability in both blood and PLF (P<0.01), significantly decreased serum TNF-α and IL-6 levels, significantly increased IL-10 level, and significantly alleviated lung and liver tissue injuries (P<0.01). The above parameters were not significantly different between the DMSO group and CLP group. Conclusion Andrographolide has protective effect against CLP-induced sepsis in mice, which might be associated with regulation of the immunological function, enhancement of bacterial clearance and inhibition of excessive inflammatory response during sepsis.

Abstract:Objective To investigate the effect of cellular Fas-associated death domain-like interleukin-1β-converting enzyme-inhibitory protein (c-FLIP) antisense oligonucleotide (ASODN)-loaded nanoparticles (NP) on the human orbital rhabdomyosarcoma xenograft in nude mice, so as to assess the feasibility of nanoparticles as a gene vector. Methods The model of human orbital rhabdomyosarcoma xenograft was established in nude mice, and the tumors were injected with c-FLIP ASODN NP, c-FLIP ASODN or normal saline (NS). The tumor volume and histopathological changes of tumor were observed. Western blotting analysis and immunohistochemical analysis were used to examine the expression of c-FLIP in tumor tissues of each group. Apoptosis of tumor cells was detected using TUNEL method. Results The growth of human orbital rhabdomyosarcoma in nude mice was significantly inhibited in ASODN NP group compared with the other two groups. Western blotting analysis showed that c-FLIP protein expression in ASODN NP and ASODN groups was significantly decreased compared with NS group (P<0.05). Immunohistochemical study showed that c-FLIP expression was found in the endochylema, and the c-FLIP positive cells in ASODN NP group was significantly less than those in the other two groups (P<0.05). Tumor cell apoptosis was observed in both ASODN NP and ASODN groups, with more found in the former, and only a few apoptotic cells were found in the NS group. Conclusion c-FLIP ASODN NP can effectively inhibit the growth of human orbital rhabdomyosarcoma xenograft in nude mice, indicating that nanoparticles may serve as a safe and effective vector for ASODN.

Abstract:Objective To observe the effect of exercise combined with alendronate sodium tablets (Aln) on receptor activator of nuclear factor kappa B ligand (RANKL)/osteoprotegerin (OPG) expression and mitogen-activated protein kinase (MAPK) signaling pathways in ovariectomized (OVX) rats. Methods A total of 90 6-month-old female SD rats were evenly randomized into sham group, OVX group, Aln treatment (OVX+Aln) group, exercise therapy (OVX+EX) group, and Aln combined with exercise treatment (OVX+Aln+EX) group. Eight weeks after ovariectomy, rats in the latter three groups were treated with intragastric 1 mg/(kg·d) Aln and/or exercise. After 12 weeks of treatment, dual-energy X-ray absorptiometry was used to examine the bone mineral density (BMD) of the left and right femurs. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of RANKL and OPG. Real-time PCR was used to examine RANKL and OPG mRNA expression in bone tissues. Western blotting analysis was used to examine the phosphorylation of c-Jun N-terminal kinase (JNK), p38 MAPK (p38), extracellular-regulated kinase (ERK), and c-Fos protein expression in bone tissues. Results The following results were observed after 12-week treatment. (1) Compared with sham group, the BMD values of the left and right femurs were significantly decreased in OVX group (P<0.05). The serum RANKL level was significantly increased, serum OPG level was significantly decreased, and the RANKL/OPG ratio was significantly increased (all P<0.05). The RANKL mRNA expression was significantly increased and OPG mRNA expression was significantly decreased in bone tissues (all P<0.05). (2) Compared with OVX group, the BMD values of left and right femurs of OVX+Aln, OVX+EX and OVX+Aln+EX groups were significantly increased (all P<0.05). The serum RANKL level was significantly decreased, serum OPG level was significantly increased, and the RANKL/OPG ratio decreased (all P<0.05). The RANKL mRNA expression was significantly decreased, and OPG mRNA expression was significantly increased in bone tissues (all P<0.05). (3) The femoral BMD values, serum RANKL and OPG levels, and RANKL and OPG mRNA expression in bone tissues in OVX+Aln+EX group were significantly different from those in OVX+Aln group and OVX+EX group (all P<0.05). (4) Compared with sham group, bone tissue p-JNK and c-Fos expression in OVX group was significantly increased (all P<0.05). However, compared with OVX group, bone tissue p-JNK and c-Fos expression in OVX+Aln, OVX+EX and OVX+Aln+EX groups was significantly decreased (all P<0.05), and there were no significant differences among the latter three groups. p-p38 and p-ERK expression was similar in all groups. Conclusion Exercise combined with Aln can improve the BMD values of left and right femurs in OVX rats, which may be related to the regulation of RANKL/OPG balance and inhibition of JNK phosphorylation and c-Fos expression.

Abstract:Objective To explore the neuroprotective effects of edaravone in 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP)-C57BL/6J-Parkinson’s disease (PD) model mice and the related mechanisms. Methods Totally 90 male C57BL/6J mice were evenly randomized into edaravone (ED) group, PD model group and normal saline (NS) group. Subcutaneous injection of MPTP was used to make PD model, and ED group was then administered with ED(3 mg/kg). Rotarod number was detected by rotarod test. Tyrosine hydroxylase-immunoreactive (TH-ir) neurons expression in the substantia nigra (SN) of mice was observed by immunohistochemistry staining. Brain-derived neurotrophic factor(BDNF) mRNA and protein expression in the SN of mice were tested by RT-PCR and Western blotting analysis, respectively. Results Compared with NS group, rotarod numbers in ED and PD groups were significantly less (P<0.05, P<0.01), the number of TH-ir neurons in SN was significantly reduced (P<0.05, P<0.01), and BDNF mRNA (all P<0.01) and protein (all P<0.05) expression was significantly decreased. Compared with PD mice, rotarod number in ED mice was significantly more (P<0.05), the number of TH-ir neurons in the SN was significantly increased (P<0.05), and BDNF mRNA (P<0.01) and protein (P<0.05) expression was significantly enhanced. Conclusion ED can increase the expression of BDNF mRNA and protein in the SN of C57BL/6J-PD model mice, alleviate MPTP damage, and has a protection effect on dopaminergic neurons.

Abstract:Objective To determine the median effective effect-site concentration (EC₅₀) of remifentanil required to inhibit hemodynamic responses induced by intubation during target-controlled infusion (TCI) of propofol in patients with Parkinson’s disease. Methods A 11 matched case control design was used in the study. Thirty-one ASAⅠ-Ⅱ patients with Parkinson’s disease undergoing general anesthesia were enrolled, and 31 controls were the hospitalized patients with non-Parkinson’s disease during the same period (same sex, age ±3 years old). A TCI of propofol was used to maintain a predetermined effect-site concentration of 3 μg/mL. The target effect-site concentration of remifentanil was determined by a modified Dixon’s up-and-down sequential method. The initial target effect-site concentrations of remifentanil were 3 ng/mL in the two groups and the increments or decrements between the two consecutive patients were 0.2 ng/mL. A positive response meant that either the heart rate or the systolic blood pressure was elevated by 15% in the first 5 min after intubation. Results There was no patient showed sign of muscle rigidity, and no patient recalled awareness during anesthesia in postoperative follow-up. During TCI of propofol, the EC₅₀ of remifentanil required for inhibiting intubation induced hemodynamic responses for patients with Parkinson’s disease was 2.20 ng/mL (95% confidence interval: 1.86-2.60 ng/mL). And the EC₅₀ of remifentanil required for control group was significantly higher than for patients with Parkinson’s disease (2.93 ng/mL, 95% confidence interval: 2.72-3.15　ng/mL). Conclusion When combined with TCI of propofol (target effect-site concentration 3 μg/mL), the EC₅₀ of remifentanil required for inhibiting intubation induced hemodynamic responses for patients with Parkinson’s disease is less than that for non-Parkinson’s disease.

Abstract:Objective To investigate the prognostic factors for poor postoperative outcomes of patients with dural arteriovenous fistulas (DAVFs). Methods The clinical data of 153 patients with DAVFs, who were treated in our institute over the past 6 years, were retrospectively analyzed. The clinical manifestations, angioarchitecture, treatment methods, neuroradiological results, and clinical outcomes were collected for Kaplan-Meier estimation and Cox regression analysis. Results The patients were followed up for a mean of (38.1±16.3) months. Twenty-one patients showed modified Rankin scale (mRS) 3-6 during recent follow-up; 4 patients died of postoperative complications, another 8 patients died during follow-up, and 9 patients presented with permanent neurologic defects. Kaplan-Meier estimation suggested that patients with poor preoperative neurologic status (mRS≥3), venous sinus thrombosis, combined arteriovenous approach embolization, partial obliteration, parital obliteration with cortical venous reflux (CVR), and postoperative restriction of dominant venous sinuses tended to have poor clinical outcomes. Cox regression analysis demonstrated that poor preoperative neurologic status (mRS≥3) (P=0.018), partial obliteration with CVR (P=0.001), and postoperative restriction of dominant venous sinuses (P=0.000 1) were risk factors predicting poor postoperative outcomes. Conclusion Poor preoperative neurologic status, partial obliteration with CVR, and postoperative restriction of dominant venous sinuses are the independent risk factors for poor clinical outcomes of DAVFs.

Abstract:Objective To detect the single nucleotide polymorphisms of rs1695 of glutathione S-transferase P1\[GSTP1 (rs1695)\] in the peripheral blood of breast cancer patients, and to analyze its association with the efficacy of paclitaxel/anthracycline-based chemotherapy in stage Ⅳ breast cancer patients. Methods The genotypes of GSTP1 (rs1695) gene polymorphisms were determined by polymerase chain reaction (PCR)-high resolution melting (HRM) method in 128 cases of female stage Ⅳ breast cancer, and 10% samples were tested by gene sequencing technique according to the PCR-HRM results. The SPSS 11.5 software was used for statistical analysis, the Hardy-Weinberg equilibrium test was used for genetic equilibrium distribution of genotypes, the correlation of different genotypes with chemotherapy responses was analyzed by the χ2 test and Fisher’s exact test, and non-conditional logistic regression model was used to calculate odds ratio (OR) and 95% confidence interval (CI). Results Of the 128 cases of stage Ⅳ breast cancer, GSTP1(rs1695) AA genotype accounted for 57.8% (74/128), AG genotype for 39.8%(51/128), and GG genotype for 2.3%(3/128). Hardy-Weinberg test suggested that our research had a group representation (P>0.05). Chemotherapy with paclitaxel/anthracycline yielded an efficiency rate of 57.03% (73/128) and an inefficiency rate of 42.97% (55/128), and in the latter the stable disease accounted for 18.75% (24/128). Patients carrying GSTP1 (rs1695) AG/GG genotype had a better response to chemotherapy than those carrying AA genotype (OR=4.139, 95%CI:1.907-8.975,P<0.01), and GSTP1(rs1695) G gene carriers had a better response than A gene carriers (χ²=12.163,P<0.01). Among the 70 cases receiving combined treatment with anthracycline, those carrying GSTP1 (rs1695) AG/GG genotype had a better response than those carrying AA genotype (OR=4.016, 95%CI:1.404-11.483, P<0.01), and GSTP1(rs1695) G gene carriers had a better response than A gene carriers (χ²=5.943,P<0.05). Conclusion Genetic polymorphisms in GSTP1 (rs1695) can serve as a genetic marker to forecast the chemotherapy response of stage Ⅳ breast cancer patients to paclitaxel/anthracycline-based chemotherapy, which is worthy of further large sample study.

Abstract:Objective To investigate the in vitro effect of xylitol on the proliferation and acid production of oral Candida, and so as to explore new prophylaxis methods against oral fungal infection. Methods Pathogenic oral Candida were clinically harvested, cultured and purified in laboratory from the saliva of 100 patients wearing dentures. The proliferation and acid production of 4 types of commonly-seen Candida albicans (C.albicans), Candida glabrata, Candida tropicalis and Candida krusei, were observed in Sabouraud’s glucose medium, with glucose replaced by different concentrations of xylitol (ratios of glucose and xylital being 41, 32, 23, 14, and 05), so as to evaluate the inhibitory effect of xylitol on the growth of different kinds of Candida. At the same time, the proliferation and acid production of C. albicans were compared when cultured with different concentrations (5%, 10%, 15%, and 20%) of xylitol (experimental groups) for 6, 12, 24, 48, and 72 h under aerobic and anoxic conditions. Results The D₆₀₀ values of four kinds of Candida in pure xylitol culture media were significantly lower and pH values were significantly higher compared with those in the glucose combined with xylitol culture media (P<0.05). Under aerobic condition, compared with the group without xylitol, the pH values in the experimental groups with different concentrations of xylitol were significantly higher and the D₆₀₀ values were significantly lower (P＜0.05), except for the D₆₀₀ values at 48 and 72 h with 5% xylitol, D₆₀₀ value at 48 h with 10% xylitol, and pH value at 48 h with 5% xylitol. Under anoxic condition, D₆₀₀ values were all significanlty increased and the pH values were all significantly decreased at all time points in the experimental groups (P＜0.05). Conclusion Xylitol can significantly inhibit the proliferation and acid production of the four common Candida species, suggesting that xylitol may serve as a substitute of sugar and inhibitor of pathogenic Candida and may play an important role in preventing and controlling oral infections of pathogenic Candida.

Abstract:Objective To generate Rtn4-A/B knockout mouse model and to explore the biological function of the Rtn4-B gene. Methods The targeting construct for inactivating Rtn4-A/B gene was prepared by bacterial artificial chromosome (BAC). The vector was linearized and electroporated into 129SvEv mouse embryonic stem cells (ES cells). Then the Rtn4-A/B knockout ES cells were microinjected into blastula of C57BL/6J mice after superovulation. F1 hybrid mice were bred to obtain mouse aggregation chimeras, and were identified by PCR amplification of tail genomic DNA. Results Fourteen clones of gene-targeted ES cells were identified after gene knockout and five male chimeras with a higher than 50 chimeric ratio were produced after microinjection into the blastula. Finally four Rtn4-A/B hybrid mice were obtained. Conclusion A Rtn4-A/B deficient mouse strain has been successfully generated by homologous recombination using genetically modified ES cells.

Abstract:Glucocorticoid (GC) has important physiological and pharmacological effects, which are mainly mediated by the GC receptor (GR). As a ligand-dependent transcriptional factor, GR activity is regulated by phosphorylation as well as GC; it is subject to hormone-dependent and -independent phosphorylation on several serine and threonine residues, especially in the N terminus. The GR is phosphorylated by cell-specific kinases such as cyclin-dependent kinases (CDKs), glycogen synthase kinase 3β (GSK3β) and mitogen-activated protein kinases (MAPKs). Phosphorylation regulates signaling and transcriptional activity of GR, thereby modulates the response of cells to GC and may be involved in the development and progression of diseases. Here we reviewed the recent research on GR phosphorylation and its pathophysiological significance.

Abstract:The damage of chronic kidney disease-mineral and bone disorder (CKD-MBD) has drawn increasing attention in recent years. In addition to traditional pathogenic factors, energy regulatory factors such as leptin, insulin, and adiponectin also participate in the development and progression of CKD-MBD. Hyperleptinaemia can directly or indirectly affect bone formation. CKD patients with diabetes have injured osteoblast function and are liable to have low dynamic osteopathy with low parathyroid hormone (PTH). Adiponectin is increased in CKD patients, which can be used as a marker for severity of osteopathy. More basic and clinical researches are needed to elucidate the mutual influence between CKD-MBD and energy metabolism. The interventions targeting molecular mechanisms may bring new treatments to CKD-MBD. This paper reviewed the mutual influence of CKD-MBD and energy metabolism.

Abstract:Objective To construct novel recombinant baculoviruses with Tet-On system and enhanced green fluorescent protein (EGFP) or hepatic growth factor (HGF) which could be regulated by different concentrations of doxycycline (DOX). Methods The recombinant plasmids pFast-Tet, pTRE-EGFP and pTRE-HGF were digested. The target fragments were collected and connected to pFast-Tet, the resultants were used to transform DH10Bac competent cells containing AcMNPV Bacmid and helper plasmid, and the Bacmid DNA were identified (named Ac-EGFP and Ac-HGF) after selection and extraction. Ac-EGFP and Ac-HGF were then transfected into bone mesenchymal stem cells (BMSCs), and the expression of EGFP and HGF were regulated by different concentrations of DOX (EGFP:0, 200, 500, and 1 000 ng/mL; HGF: 0, 10, 100, 500, 1 000, and 1 200 ng/mL); EGFP expression was observed under fluorescence microscope and the level of HGF expression was detected by ELISA. Results It was verified that Tet-On system was successfully constructed in a baculovirus vector with EGFP or HGF, and they were highly transfected into BMSCs. EGFP and HGF were highly expressed when exposed to high concentrations of DOX. And the expression of EGFP and HGF were gradually decreased at low concentration or absence of DOX. Conclusion Tet-On system can be used to construct a new recombinant baculovirus vector containing EGFP or HGF and it can stably and highly transfect BMSCs; different concentrations of DOX can lead to different expression of EGFP and HGF, and they are in low background expression without DOX.

Abstract:Objective To investigate the possible role of wild-type p53-induced phosphatase 1 (Wip1) in repairing radiation-induced DNA damage in nucleus pulposus (NP) cells, so as to provide reference for treatment of intervertebral disc degeneration. Methods Human NP cells were cultured in vitro and the expression of Wip1 was knocked down by small interfering (siRNA) technology; the NP cells were exposed to radiation (4,10,15 and 25 Gy). DNA damage and repair were observed by comet assay, and the results were compared with NP cells not treated by siRNA. Co-immunoprecipitation (Co-IP) was used to detect the potential binding protein of Wip1, and based on the screening results, qRT-PCR was used to examine Wip1 expression and expression of potential binding protein. Results Comet assay showed that the repair of DNA damage in Wip1 knockdown NP cells was not affected when exposed to 25 Gy radiation, but DNA damage repair was persistently reactivated. The marker molecules of DNA damage repair were not detectable 24 h after radiation in the control group, but they could be detected 48 h after radiation in Wip1 knockdown group. Co-IP results showed that the distribution of Wip1 and Bmi1 coincided in the nuclei of normal cells exposed to radiation, aggregating at the DNA damage site, and the coincidence disappeared after knockdown of Wip1. qRT-PCR results showed that, compared with the normal intervertebral tissues, degenerated intervertebral tissues had significantly decreased Wip1 and Bmi1 expression, and the expression of Wip1 was positively correlated with Bmi1 expression (P<0.05). Conclusion Wip1 plays an important role in the DNA damage repair of NP cells by regulating DNA damage repair phase, and Bmi1 may also participate in this process.

Abstract:Objective To develop a rat model of periapical periodontitis caused by repeated root canal infection with Enterococcus faecalis (E.faecalis), and to evaluate the disease progression by observing the periapical lesion area, inflammation, and TNF-α expression in the root apex. Methods Thirty Sprague Dawley rats were used in this study. The pulps of the bilateral first maxillary molar teeth of each rat were exposed, absorbent cotton with lipopolysaccharides (LPS) was implanted, and then the teeth were exposed in the oral environment for four weeks to induce periapical periodontitis of primary root canal infection with mixed bacteria. Then the root canal was subjected to disinfection treatment for two weeks, followed by inoculation with E.faecalis suspension for one week to produce repeated root canal infection model with E.faecalis for a 3-week observation. X-ray, H-E staining and immunohistochemical staining were used to observe the periapical lesion area, inflammation, and TNF-α expression in the root apex at one week, two weeks, three weeks after establishment of repeated root canal infection model; the above parameters were also observed for periapical periodontitis caused by primary root canal infection with mixed bacteria (6 weeks later) and for root canal disinfected by Ca (OH)₂(for 6 weeks). Results (1) Periapical periodontitis caused by repeated root canal infection with E.faecalis had gradually increased periapical lesion area, with unclear damage border; the infection reached a severe level during the 1-2 weeks and became chronic inflammation at week three; and the expression of TNF-α began to decrease after two weeks. (2) The periapical periodontitis caused by primary root canal infection with mixed bacteria had a smaller periapical lesion area, with clear damage border, slighter infiltration of inflammatory cells, and less TNF-α expression. (3) The periapical periodontitis with root canal disinfected by Ca (OH)₂ had the smallest periapical lesion area, with disappeared inflammation, slight TNF-α expression, and hyperplasia of cementum and alveolar bone. Conclusion E.faecalis has a strong periapical damaging effect, with continuous bone absorption observed even under the chronic inflammatory status.

Abstract:Objective To retrospectively analyze the intraoperative complications of laparoscopic radical prostatectomy (LRP) during the initial application, so as to analyze the reasons and summarize the experience. Methods The videos of the 60 patients undergoing LRP for prostate cancer in our hospital between January 2010 and November 2012 were reviewed to analyze the intraoperative complications. The patients were the first 20 cases done by 3 surgeons independently. The clinical data, tumor staging, and other related factors were collected for analyzing the reasons for complications of LRP. Results Complications were observed intraoperatively in the 23 patients. The most frequent complications included mistaken suture of bladder catheter (7 cases), open conversion due to intraoperative bleeding (5 cases), residual prostate tissue (5 cases), secondary anastomosis (4 cases), rectal injury (1 case), and specimen missing (1 case). All the events were managed during the operation. The reasons for the complications included unclear anatomical structure, unclear surgical landmark, poor cooperation with assistants, and inconsiderate operative details. Some patients were in the advanced stage, making it difficult to perform the operation. Conclusion The patients with low surgical risk should be chosen for the initial application of LRP, and a stable operation team should be formed. A standard strategy should be formed regarding the equipment selection, surgical procedure, anatomic marking, and manipulation based on viewing the operation; and improvement should be made gradually in practice.

Abstract:Objective: To study the clinicopathologic characteristics, survival time and prognostic factors of gastric cancer（GC） in young patients. Methods: The authors studied 1135 patients with GC who received curative resections. They were classified into two groups: those aged 40 years or less (young); and those aged more than 41 years (elderly). Clinicopathologic characteristics, survival rates and prognostic factors were analyzed. Results: Young patients were predominately female, had more diffuse type, and had lower proportion of tumors in the upper third of the stomach. However, survival rate was not significantly different between the two groups. TNM stage, number of lymph node metastasis were independent negative prognostic factors in young patients. Conclusions: GC in young patients demonstrates only minor deviations from the older with a similar survival rate, which suggests positive and standard treatment should be taken to young patients with GC.

Abstract:Aim To investigate the correlation between between military physical training and body fat percentage , lean body mass percentage, muscle percentage, skeletal muscle percentage and BMI. Methods A total of 5631 male graduating students and 558 new students from 16 different military academies were involved in the study to conduct the body composition testing and analyze the correlation between physical training and the indicators by using literature review and mathematical statistics. Results The body fat percentage in the male graduating students were significantly lower than that in the new students(P=0.0036）. While the lean body mass percentage, muscle percentage, skeletal muscle percentage and BMI in the male graduating students were significantly higher than those in the new students（P=0.0037、0.0020、0.0411 and <0.0001, respectively ）.Conclusion A long-term planned, targeted and military physical training program can significantly reduce body fat percentage, increase lean body mass percentage, muscle percentage, skeletal muscle percentage and BMI and enhance students' physical fitness.