Objective To investigate the mechanism of the radiosensitivity effect of Cox-2 gene in esophageal cancer.Methods Cox-2 specific siRNA was constructed and transfected to EC9706 cells to downregulate intracellular Cox-2 expression.The expressions of MMP-2,Bcl-2 mRNA,AKT and phosphorylated AKT proteins were assayed after radiation.Colony formation,cell proliferation,apoptosis and cell invasion in vitro were examined as well.One-way ANOVA method was used to analyze the data.Results Affter 2 and 4 Gy irradiation,a significant increase in the mRNA expression of Bcl-2 was observed in the Cox-2 up-regulation group (F =3.36,4.32,P ＜ 0.05).In the group of Cox-2 downregulation,the expression of MMP-2 mRNA was significantly reduced(F =3.86,8.09,P ＜ 0.05).Affter irradiation,a significant decreaseof Bcl-2 mRNA (F =3.73,5.64,P ＜ 0.05) as well as an increase of Bax(F =7.03,7.42,P ＜ 0.05) was detected,and the levels of total and phosphorylated AKT proteins had the highest level in the Cox-2 upregulation group and had the lowest level in the Cox-2 downregulation group.In the Cox-2 downregulation group,the apoptosis induction obviously increased with dose (F =317.40,P ＜ 0.05),and the proportion of cells in Go-G1 phase gradually increased but the proportion of cells in S and G2-M phases decreased,concomitant with the obvious suppression of cell proliferation,in addition,cell invasion was decreased.Conclusions Downregulation of intracellular Cox-2 mRNA expression,concomitant with subsequent downregulation of MMP-2 and Bcl-2 and upregulation of Bax,resulted in reduction of the invasion and metastatic capabilities of tumor cells,and induction of Go-G1 phase arrest and apoptosis.Downregulation of AKT and phosphorylated AKT (pAKT) protein expression might also interfere with the capability of the PI3K/Akt signal transduction pathway to resist radiotherapy.

AIM: To investigate the influence of Buxinqi Capsule (BXQ) on myocardial ischemia reperfusion injury in rats. METHODS: In this study, the experimental model was established by reperfusion for 60 minutes in rats after ligaturing left anterior descending coronary artery (LAD) for 30 minutes. Serum creatine phosphokinase (CPK), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyodebis (MDA), area of myocardial infarction and occurrence of arrhymia were investigated. RESULTS: BXQ significantly decreased level of CPK and LDH and MDA, and obviously improved the activity of SOD, decreased reperfusion arrhythmias and arrhythmias severity index (ASI), and decreased the area of myocardial infarction. CONCLUSION: BXQ has protective effect on the damage of myocardia ischemia reperfusion in rats

ObjectiveTo investigate the effects of using transforming growth factor-β3 (TGF-β3) combined with dental pulp stem cells (DPSCs) on the regeneration of rabbit facial nerve.MethodsSixteen New Zealand adult rabbits were selected randomly.These rabbits were divided into three groups (TGF-β3 and DPSCs treated group,TGF-β3 control group and PBS control group.treatment group (silicone guidance channel with collagen protein sponge were filled with TGF-β3 and DPSCs),TGF-β3 control group (silicone guidance channel with collagen protein sponge were filled with TGF-β3) and PBS control group(silicone guidance channel with collagen protein sponge were filled with PBS).After 3 months,a series of examinations were performed,including gross morphology,histological staining,neuroelectrophysiological tset.ResultsIn the experimental group,the diameter of the regenerating nerve and near distal nerve stem were almost the same.There were no formation of neuroma.The adventitial angiogenesis was rich and with tough texture.3 months after operation,in the experimental group,the facial nerve membrane integrity,nerve fibers arranged in neat rows,form a more complete,myelin swelling.The total number of regeneration of nerve fibers in the experimental group were more than of control groups,statistical analysis was significant (P＜0.05).Diameter of regenerating nerve fibers in the experimental group were greater than that of control groups,statistical analysis was significant (P＜0.05).The ultra-thin section showed that the regenerated fibers in the treatment group were mainly myelinated never fiber.The layer structure of myelin sheath was clear,and there were rich organells axoplasma.The neuroelectrophysiological examinations revealed that the latency of nerve and muscle action conduction in the treatment group was shorter than that of the control groups,the treatment group:(1.96±0.32) ms,the TGF-β3 control group:(2.35±0.41) ms,the PBS control group:(3.42±0.55) ms.The wave amplitude of nerve and muscle action conduction in the treatment group was obviously higher than that of control groups,the treatment group:(11.06±3.25) mV,the TGF-β3 control group:(8.40± 1.68) mV,the PBS control group:(4.62±0.77) mV.ConclusionThe combination of TGF-β3 and DPSCs can improve the effects on the repair of facial nerve injury.

Objective To investigate the feasibility of combing the application of transforming growth factor-β3 (TGF-β3) with concentration of 100 ng/μl and dental pulp stem cells (DPSCs) for recovering rabbit facial nerve transverse trauma.Methods Thirty-six healthy adult Zelanian rabbits of clean-grade were selected and randomly divided into group DPSCs+TGF-β3 (experimental group), group TGF-β3 (control group 1) and group PBS (control group 2) with 12 rabbits in each group.The operations for all three groups were applied at rabbit's left cheek.A model of traumatic transection was set on upper buccal branch, then 100 ng/μl TGF-β3 solution and 0.1 ml of 1 ×108/L DPSCs suspension were added into regeneration chamber for the experimental group, while the same amount of 100 ng/μl TGF-β3 solution was added for group TGF-β3 and the same amount of PBS for group PBS.The recovery of facial nerve regeneration with the prepared animal's specimen was evaluated in the 1st, 4th and 12th week after the operation on sacrificed rabbits.Results The effects on nerve regeneration recovery for the experimental group was superior to that of control groups 1 and 2 with all the 36 models included in the result analysis, and that of control group 1 was superior to that of control group 2, which was getting better with the extension of time.Conclusions The combined application of TGF-β3 and DPSCs can effectively promote the facial nerve regeneration, which is better than that of single application of TGF-β3.Meanwhile, the effect with TGFβ3 application is better than that with application of PBS.

Objective To assess mucin gene expression in Barrett's esophagus.Methods Mucin core protein-MUC1,MUC2,MUC3,MUCSAC and MUC6 were detected by immunohistochemistry.The re- lationship between mucin expression and magnification-endoscopic characteristics,pathohistologic epithelial types of Barrett's esophagus was analyzed.Results Mild expression of MUC1 was predominantly found in the superficial epithelium of both gastric and specialised intestinal metaplasia.In a small number of specimens, mild expression of MUC1 was also noted in glands.Strong MUC2 expression was noted only in the goblet cells in Barrett's oesophagus.MUC3 was expressed in the superficial columnar cells of specialized intestinal metaplasia with or without globlet cells but not in gastric metaplasia of the oesophagus.In some specimens MUC3 was expressed in the vacuolus of the globlet cells and the lumen of gland.Strong staining of MUCSAC was noted in the columnar epithelium of both gastric metaplasia and specialized intestinal metaplasia in Barrett's oesophagus,as well as expressed in the cytoplasm and vacuolus of the globlet cells in some speci- mens.Expression of MUC6 protein was detected at the basement of the crypts in gastric metaplasia and spe- cialised Barrett's glands.Expression of MUC2 and MUC3 protein was found much higher in villous or irregu- lar pit pattern than that in dot or rod pit pattern(P

This study was aimed to compare the effects of acupuncture stimulation on different acupoints, which included LR14-Qimen, ST25-Tianshu, GB34-Yanglingquanand CO11-pancreas and gallbladder (auricular vagus nerve intensively distributed areas) on gallbladder pressure and Oddi sphincter function of guinea pig, in order to discuss acupuncture effect on different acupoints in the regulation of motor function of extrahepatic biliary system. A total of 20 adult guinea pigs of both sexes were randomly divided into two groups, with 10 in each group. The detection was made on changes of Oddi sphincter electromyography and gallbladder pressure before and after acupuncture. A double-hook electrode was implanted into the Oddi sphincter for the detection of Oddi sphincter electromyography. And one small artificial balloon was put into the gallbladder to measure the intrabiliary pressure. After the guinea pig was anesthetized, changes of gallbladder pressure as well as the electromyography frequency and amplitude index of Oddi sphincter of the normal guinea pig were recorded. When the guinea pig was stabilized, acupuncture was given on LR14, ST25, GB34 and CO11 for 1 min. Observations were made on acupuncture effects of gallbladder pressure and Oddi sphincter function of guinea pig. The results showed that after acupuncture on LR14 and ST25, the electromyography frequency of Oddi sphincter was obviously increased compared with before acupuncture (frequency:P < 0.01, amplitude:P < 0.05); the gallbladder pressure was obviously decreased (P < 0.05). After acupuncture on CO11 and GB34, the electromyography frequency and amplitude index of Oddi sphincter were obviously decreased compared with before acupuncture (P < 0.05); the gallbladder pressure was obviously increased (P < 0.01). It was concluded that acupuncture on LR14 and ST25 can obviously inhibit the movement of gallbladder and promote the myoelectricity release of Oddi sphincter. Acupuncturing on CO11 and GB34 can apparently promote the shrinking movement of gallbladder and inhibit the myoelectricity release of Oddi sphincter.

Objective To explore the effect of long-term depleted uranium (DU)ingestion on testosterone production in rats, and its involvement mechanism. Methods Male and female rats (F0 and F1 respectively) for 160 days, respectively. The contents of testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) in serum were detected in 20 months of F0 generations, and 15 months of F1 generations. RT-PCR was used to analyze the levels of StAR mRNA and P450scc mRNA. Results Compared with the normal control group, the testosterone contents in exposed F0 and F1 generations increased, the lowest was 51.73 U/L, but those of LH and FSH decreased. The expression of StAR mRNA in the low-doze group of F1 generation (StAR/β-actin = 1.35) was up-regulated, down-regulated for other groups.compared with the normal control group (P450scc/β-actin = 0. 313), the expression of P450scc mRNA in the low- and high-dose groups of F0 generation were decreased (P450scc/β-actin = 0.21), and those in the low- and high-dose groups of F1generation were increased (P450scc/β-actin = 0.623) (P ≤ 0.01). Conclusion Long-term DU exposure inhibit the male reproduction by intervening the sexual hormone production through down-regulated the expression of StAR mRNA and P450scc roRNA.

Objective To explore the changes of renin-angiotensin-aldosterone system in pediatric severe sepsis treated with continuous blood purification(CBP).Methods Prospective study,35 cases of critically ill children diagnosed with severe sepsis and admitted to PICU of Shanghai Children's Hospital of Shanghai Jiaotong University from June 2012 to May 2014 served as reseach objective.Based on the monitoring of vital signs,including central venous pressure,arterial blood pressure,mean arterial pressure,patients were treated with conventional therapy,as antibiotics,fluid therapy,and CBP by mode of continuous veno-venous hemodiafiltration or high volume hemofiltration.Plasma levels of rennin activity,angiontensin Ⅱ and aldosterone were measured by radioimmunoassay before and 24 h after CBP.Twenty-five cases of blood samples taken from the children collected from health care for liver function examination were matched as control group.Results Plasmalevelsofrenninactivitywere(2.11 ±1.93) pg/(L·h),(1.27±1.56) μg/(L·h),(0.37 ± 0.22) μg/(L· h) before and 24 h after CBP and control group,respectively.The levels of angiontensin Ⅱ were (426.78 ±332.37) ng/L,(364.40 ± 325.51) ng/L,(41.70 ± 10.81) ng/L,respectively.And the levels of aldosterone were (255.12 ± 218.18) ng/L,(134.92 ± 104.13) ng/L,(106.88 ±43.18) ng/L,respectively.The plasma levels of rennin activity,angiontensin Ⅱ,and aldosterone were higher in sepsis cases than in control group,while decreased obviously after CBP treatment(P ＜0.01,P ＜0.05).Eleven cases died and mortality was 31.4％ (11/35).After 24 h of CBP,the mean arterial pressure improved in 26 cases with septic shock and dopamine dose reduced(P ＜ 0.01).Conclusion The reaction of renin-angiotensin-aldosterone system is increased significantly in pediatric severe sepsis.CBP can down-regulate the levels of rennin activity,angiotensin Ⅱand aldosterone,but not worsen the circulation function.

OBJECTIVE To investigate enhanced immune function of methionine encephalin (MENK) and its anti-tumor mechanism in CT26 colon cancer mouse model. METHODS 3×106 CT26 cells were implanted subcutaneously in BALB/c mice. Four days after, MENK was peritoneally administrated at the concentration of 20 mg·kg-1 for 14 d. The percentage of MDSCs in bone marrow, spleen, blood, tumor and liver were detected by flow cytometry. Non- esterified fatty acid (NEFA), triglycerides (TG) and total cholesterol (T-CHO) in liver homogenate were tested by a NEFA test kit, a TG test kit and a T- CHO test kit respectively. qRT- PCR and Western blot were used to measure mRNA and protein levels of inflammation-, glycometabolsim- and lipometabolsim-associated indexes in liver. RESULTS MENK decreased percentages of MDSCs in bone marrow, spleen, blood and tumor in colon cancer mice. MENK-treated mice displayed elevated ratio of CD4+T and CD8+T cells in spleen as well as increased T and B lymphocytes proliferation. Meanwhile, MENK also ameliorated liver damage reflected by lower levels of GPT and GOT in serum and reduced risks of cancer- associated index including inflammation, high lipid and high glucose. Furthermore, MENK lowered down the levels of NEFA, TG and T- CHO in liver homogenate. MENK treatment decreased expression of p- STAT3, increased expression of p-AKT, IRS1 and Glut4 at protein level as well as reduced lipogenesis-associated genes and elevated glycolysis-associated genes in liver of tumor bearing mice. Also, abated expression of genes associated with MDSCs generation (M-CSF, GM-CSF, IL-6, IL-1β) and migration (S100A9, KC) was observed within shrunken subcutaneous tumor by MENK intervention. CONCLUSION MENK has the ability to strength immune function against colon cancer by reducing MDSCs and improving liver metabolism.

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.