Objective To investigate the effects of penehyclldine hydrochloride postconditioning on hind limb ischemia-reperfusion (I/R) injury in rats. Methods One hundred and forty-four male Wistar rats weighing 220-250 g were randomly divided into 4 groups (n = 36 each): control group (group C), I/R group, ischemic postconditioning group (group IPO) and penehyclidine hydrochloride postconditioning group (group PHPO). Limb ischemia was induced by occlusion of bilateral hind limb for 3 h followed by 24 h reperfusion. In group IPO, the animals were subjected to three cycles of 30 s reperfusion and 30 s ischemia on the proximal part of the hind limb,while in group PHPO, the animals received iv injection of penehyclidine hydrochloride 0.15 mg/kg (in normal saline 1 wl) via the caudal vein immediately after reperfusion. Blood samples from the inferior vena cava were taken for determination of activities of serum LDH and CK and concentrations of TNF-α and IL-10 at O, 1, 3, 6, 12 and 24 h of reperfusion. Then 6 rats were sacrificed at each time point respectively and the quadriceps femoris tissues of the hind limb were removed for determination of MDA content, activities of SOD, MPO, Na+ -K+-ATPase and Ca2 + - Mg2+ -ATPase, and HIF- 1 α expression and microscropic examination. Results Compared with group C, the activities of serum LDH and CK and concentrations of TNF-α and IL-10 were significantly increased, the activities of SOD, Na+-K+-ATPase and Ca2+-Mg2+-ATPase were decreased, MPO activities and MDA content were increased, HIF-1α expression was up-regulated in group I/R, IPO and PHPO (P ＜0.05 or 0.01). Compared with group I/R, the serum activities of serum LDH and CK and TNF-α concentration were significantly decreased, IL- 10concentration was increased, activities of SOD, Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase were increased, and MPO activity and MDA content were decreased, HIF-1α expression was down-regulated (P ＜ 0.01), and the pathologic changes was reduced in group IPO and PHPO. Compared with group IPO, the serum LDH activity and TNF-α concentration were significantly decreased, CK activity and IL-10 concentration were increased, activities of SOD and MPO and MDA content were decreased and HIF-1α expression was down-regulated (P ＜ 0.05 or 0.01).Conclusion Penehyclidine hydrochloride postconditioning can reduce hind limb I/R injury in rats, and the mechanism is related to inhibiting inflammatory response and lipid peroxidation, reducing calcium overload and improving cellular energy metabolism.

Female ; Humans ; Infant, Newborn ; Mouth ; microbiology ; Pregnancy ; Premature Birth ; microbiology

Aim To study the protective effects of sphingosine 1-phosphate (S1P) postconditioning on rat myocardial cells injured by hypoxia/reoxygenation in reperfusion injury salvage kinase ( RISK ) signal path-way. Methods The cultured rat H9c2 cells were ran-domly divided into seven groups: ( 1 ) control group;(2) hypoxia/reoxygenation (H/R) group; (3) S1P group;(4) S1P+LY294002 group(S1P+LY); (5) LY group; ( 6 ) S1 P +PD98059 group ( S1 P +PD );(7) PD group. The viability of H9c2 cells was detec-ted using MTT method. The content of MDA in the cultured medium and the activity of T-SOD and Mn-SOD were measured with colorimetry. The concentra-tion of intracellular free calcium ion was detected by confocal microscopy. The rate of cell apoptosis was de-termined by flow cytometric analysis. Western blot was used to assess phosphorylation of Akt and ERK1/2 in H9c2 cells. Results Compared with the H/R group, S1P significantly increased vaibility of cells, lowered the rate of apoptosis, decreased the content of MDA in the culture medium, increased the activity of T-SOD and Mn-SOD, reduced concentration of intracellular calcium and increased the phosphorylation of Akt and ERK1/2 . When added LY294002 or PD98059 , the effects of S1P above were inhibited. Conclusion S1P protects H9 c2 cells against hypoxia/reoxygenation inju-ry. The protection of S1P was inhibited by LY294002, the inhibitor of PI3 K/Akt and PD98059 , the inhibitor of ERK1/2 . S1 P protects H9 c2 cells against hypoxia/reoxygenation injury via RISK pathway.

Objective To investigate the expressions of TLR2 and TLR4 in patients with psoriasis vulgaris and the efrect of methotrexate(MTX)on them.so as to explore the therapeutic mechanism of MTX in psoriasis vulgafis.Methods Forty-three patients with psoriasis vulgaris were recruited into the study together with 30 normal human controls.Oral MTX was given to patients with an interval of 12 hours for three times per week until the control of conditions followed by 4 weeks of mainmining treatment.The dosage of MTX was 5 mg initially and decreased to 2.5 mg in the maintaining period.Flow cytometry was used to detect the expression of TLR2 and TLR4 in peripheral blood CD14~+ cells from the controls and patients at baseline,4 and 8 weeks after the beginning of treatment.Results The expression rate of TLR2 and TLR4 in CD14~+ cells was(92.6±4.3)%and(48.5±4.6)%,respectively,in untreated patients,significantly higher than that in normal controls(botll P＜0.01).A significant increase was observed in the expression rate of TLR2 and TLR4 in patients with active psoriasis compared with those with inactive psoriasis [(97.5±4.1)%vs(87.6±5.6)%,(55.3±5.8)%vs(40.7±7.1)%,both P＜0.05].Eigh weeks after the beginning of treatment with MTX.the expression rate of TLR2 and TLR4 significantly decreased to (79.6±6.7)%and(34.6±5.9)%.respectively(both P＜0.05).The psoriasis area and severity index(PASI)score had no significant correlation with the expression rate of TLR2 or TLR4(r=0.24.0.27,both P＞0.05).Conclusions TLR2,TLR4 and innate immune response mediated by both receptors play an important role in the pathogenesis of psoriasis.MTX may exert its therapeutic effect on psoriasis by inhibiting the expression of TLR2 and TLR4.

Objective:To observe the bone marrow mesenchymal stem cell isolation,identification methods and protection mechanisms in the kidney of diabetic rats.Methods: Thirty-one rats were used to isolate bone marrow mesenchymal stem cells (BMSCs) by adherent culture method.Cell morphology was observed by phase contrast microscope.The surface molecular weight of cultured cells was detected by Real-time PCR.30 rats were induced by intraperitoneal injection of streptozotocin to establish diabetic rat model (n=15).And stem cell treatment group (n=15) according to the different treatment methods.Rats in the control group were treated with insulin combined with probucol.Stem cell treatment group was implanted with stem cells.Before and after treatment,fasting blood glucose,24 h urinary protein excretion,creatinine clearance rate and kidney weight were measured.PAS staining.And the renal tissue was detected by Western blot.Results: The primary bone marrow mesenchymal stem cells (MSCs) were cultured for 24 hours,and the cells were irregularly distributed in the culture flask.After 7 days of culture,the cells were regular,elongated,elliptical,and strong refraction,and the cells were fused with each other.The morphology of BMSCs was uniform and showed a long fusiform shape,CD44,CD31 and CD34 were observed by RT-PCR.The levels of fasting blood glucose,24 h urine protein excretion and creatinine clearance rate in the observation group were significantly higher than those in the control group (P<0.05).The results of PAS staining showed that the glomerular structure was clear,the morphology was regular,the arrangement rules of glomerular cells,the basement membrane and the encapsulation of the glomerular membrane were significantly higher than those of the control group Man′s capsule clear (P<0.05).In the control group,the glomerular volume increased,the mesangial area widened and the stroma increased significantly.The Bcl-2 protein level and the Bax and Caspase-3 protein levels were significantly decreased in the stem cell treatment group.The levels of Bcl-2 and Bax and Caspase-3 protein in the control group were significantly decreased (P<0.05).The ROS,MDA and SOD levels in oxidative stress group were lower than those in control group (P<0.05).The difference of ROS,MDA and SOD between the two groups was not statistically(P>0.05).Conclusion: It is suggested that BMSCs can be isolated from rat bone marrow mesenchymal stem cells by using adherent culture method and labeled with green fluorescent protein in vitro.Bone marrow mesenchymal stem cells can protect the kidney and inhibit the oxidative stress of diabetic kidney.

Objective To discuss the correlation between SIRT3 protein and clinicopathological parameters of gastric carcinoma.Methods Immunohistochemistry and Western blot were used to detect the expression of SIRT3 in the gastric carcinoma and normal gastric tissue.The correlation between the expression of SIRT3 and clinicopathological parameters was analyzed.Results The immunohistochemistry showed that the positive expression rate of SIRT3 protein in gastric carcinoma tissue (53.8 ％,43/80) was obviously lower than that in normal gastric tissue (86.0 ％,43/50),and the expression of SIRT3 protein showed close relationship with invasion depth,lymph node metastasis and TNM stage (P ＜ 0.05),rather than the age,gender,tumor size,or differentiation status (P ＞ 0.05).The Western blot showed that the expression rate of SIRT3 protein (SIRT3/β-actin) in gastric carcinoma tissue (0.655±0.317) was lower than that in normal gastric tissue (0.803±0.329) (P ＜ 0.05).Conclusion The expression of SIRT3 protein is lower in gastric cancer than that in normal gastric tissue,and relates to invasion depth,lymph node metastasis and TNM stage.SIRT3 may inhibit the occurrence and development of gastric cancer.

Objective To investigate the expressions of angiopoietin(Ang)-1,-2 and their receptor,Tie-2 in Patients with psoriasis vulgaris and their significance.Methods Tissue samples were collected from 30 patients with psoriasis vulgaris and 15 normal controls.Streptavidin-biotin complex(SABC)method was used to detect the expression of Ang-1,-2 and Tie-2 in these samples.Microvessel density (MVD)was assessed by CD34 staining.Results The expressions of Ang-2 and Tie-2 were significantly higher in patients with psoriasis vulgaris than in the normal controls(7.65±2.14 vs 2.54±0.91,6.09±1.97vs 1.95±0.62,both P<0.01),and higher in patients with active psoriasis than in those with inactive psoriasis (9.85±2.42 vs 4.86±1.63,8.43±1.98 vs 4.57±1.92,both P<0.05),whereas no significant difference was observed in the expression level of Ang-1 between Patients and controls or between patients with active psoriasis and those with inactive psoriasis(all P>0.05).Mcreased MVD was observed in patients、with psoriasis vulgaris compared with the controls(15.46±5.28 vs 2.68±0.79,P<0.01).Also,the expression levels of Ang-2 and Tie-2 were significantly difierent between patients with high MVD and those with low MVD(both P<0.05),while no difference was nomd in the expression of Ang-1 (P>0.05).In psoriatic lesions,a negative correlation was observed between the expression of Ang-1 and Ang-2(r=-0.521,P<0.05),and MVD was negatively correlated with the expression of Ang-2(r=0.656,P<0.01)but unrelated to the expression of Ang-l(r=0.237,P>0.05).Conclusions It is demonstrated that Ang-2 and Tie-2 closely correlate with the angiogenesis in psoriasis,and Ang-2 plays a dominant role in the regulation of angiogenesis in psoriasis.

Objective: To understand the level and risk factor of lead exposure among children in one city of Jiangsu. Methods: In northern Jiangsu Province, 373 children from 2 primary schools were enrolled and were tested for blood lead and heavy metal exposure. Lead exposure was tested in household dust of 46 children. A multivariate Logistic regression was used for lead exposure risk analysis. Spatial distribution of lead contamination in household dust was conducted and compared with the location of industrial enterprises. Results: The geometric mean of blood lead in 373 children was 25.80 mg/L,the blood lead of 3 children (0.8%) was more than 100 mg/L. Pencil biting ( OR=4.26, 95%CI=1.61-10.68, P <0.05) and lead contamination in surrounding environment ( OR=2.93, 95%CI=1.24-7.34, P =0.02) was positively related to high blood lead level in children. The geometric mean household dust lead concentrations in 46 children was 302.27 μg/mg, and household with high dust lead levels were mainly located around manufacturing enterprises. Conclusion Environmental factors correlate with blood lead level in children. Efficient strategies and public health policies are urgently needed to control and prevent environmental lead pollution. Families and schools should actively carry out health education to engourage children good hygiene habits, and effectively reduce lead exposure.

OBJECTIVETo construct a low-pH-sensing system in Streptococcus mutans (S. mutans) and to visually detect the pH in situ.

METHODSPromoter of ureaseⅠ(PureⅠ) and green fluorescence protein (gfp) DNA fragments were amplified by polymerase chain reaction (PCR) from the genome of Streptococcus salivarius 57.I and S. mutans containing the gfp fragment. The two amplified DNA fragments were ligated together and further integrated into pDL278 to construct the recombinant plasmid pDL278-pureⅠ-gfp. This recombinant plasmid was then transformed into S. mutans UA159 cells. Subsequently, the intensity of the optical density per unit area of the low-pH-sensing system was measured and compared under different pH conditions and different processing times.

RESULTSPureⅠ and gfp DNA fragments were amplified successfully with the correct molecule sizes (450 and 717 bp, respectively). The recombinant plasmid pDL278-pureⅠ-gfp was constructed and further verified by PCR and sequencing. The intensity of the optical density per unit area of the low-pH-sensing system increased with decreasing pH and increasing processing time.

CONCLUSIONSA low-pH-sensing system was constructed successfully in S. mutans. Our research verified that pureⅠ of Streptococcus salivarius can function well in S. mutans as an acid induced promoter, and provided a new method of detecting the pH of plaque biofilms in situ.

Biofilms ; Dental Plaque ; Green Fluorescent Proteins ; Hydrogen-Ion Concentration ; Plasmids ; Promoter Regions, Genetic ; Streptococcus mutans ; Streptococcus salivarius

OBJECTIVETo study the impact of neoadjuvant radiochemotherapy on erectile and urinary functions in mid-low rectal cancer patients.

METHODSSeventy mid-low rectal cancer patients from January 2012 to May 2013 in The Sixth Affiliated Hospital, Sun Yat-sen University were prospectively enrolled. According to tumor staging and patient decision, patients received neoadjuvant radiochemotherapy(50 Grays administered over a six-week period and four cycles of concomitant mFOLFOX6 chemotherapy followed by operation (study group) or surgery alone(control group). Dropouts, loss to follow up and relapse during follow-up were removed from the analysis. A total of 30 patients stayed in study group and 29 patients in control group. To assess erectile and urination functions, the five-item version of the international index of erectile function (IIEF-5) and the international prostate symptom score (IPSS) questionnaires were used before therapy and 12 months after surgery.

RESULTSIn both study and control groups, total IIEF-5 score was decreased significantly at postoperative 12-month compared to initial assessment(P<0.01). Compared with control group, IIEF-5 score change was significantly higher in study group (9.6 ± 6.1 vs. 5.3 ± 5.3; P<0.01). Total IPSS score in both groups was increased significantly at postoperative 12-month compared to initial assessment(P<0.05). No significant difference was found in IPSS score change between the two groups (3.0 ± 3.4 vs. 1.5 ± 3.0, P>0.05). Univariate analysis on study group showed that age, tumor location and maximal diameter were associated with erectile dysfunction. Age was associated with urination dysfunction (all P<0.05).

CONCLUSIONNeoadjuvant radiochemotherapy has significant impact on erectile dysfunction after surgery in mid-low rectal cancer patients.

Chemoradiotherapy ; Erectile Dysfunction ; Humans ; Male ; Neoadjuvant Therapy ; Neoplasm Staging ; Prospective Studies ; Rectal Neoplasms ; Surveys and Questionnaires ; Urinary Incontinence