Objective To investigate the intracellular signal transduction pathway of insulin promoting stem cell factor (SCF) expression in colonic smooth muscle cells (SMCs).Methods Colonic SMCs from SD rats were isolated and cultured, which were identified by α-actin immunofluorescence cytochemistry staining.SMCs were divided into 3 groups for experiment. ①Insulin concentration gradient group:Adding different insulin concentrations (0,2.5,5,20,40 and 80 mg/L) into SMCs culture and cultured for 16 hours; ② Insulin time point group:SMCs were cultured with insulin (40 mg/L) for different time durations (0,8,16,24 hours); ③ Signal pathway blocking group:SMCs were pretreated with LY-294002 or PD-98059,and then induced with insulin (40 mg/L) for 16 h.The proliferation of SMCs was examined by MTT method.The change of SCF expression was determined by Western blot and RT-PCR.Results ① Insulin at concentration of 5 mg/L remarkably promoted SMCs proliferation,and there was no significant difference in its effect with insulin concentrations at 20,40,80 mg/L (all P＜0.05).② Compared with no insulin adding group,low and medium dose of insulin (2.5,5,20 mg/L) already had a role in promoting SCF mRNA and SCF protein expression (all P ＜ 0.05). It reached maxium effect when insulin concentration was at 40 mg/L(P＜0.05),and there was no significant difference between insulin concentration at 40 mg/L and 80 mg/L (P＞0.05).③ Compared with time 0 hour,the expression of SCF increased after insulin worked on SMC (40 mg/L) for 8 hours (P＜0.05),and reached the peak after 16 hours (P＜0.05),and there was no significant difference between 24 hours and 16 hours (P＞0.05).④ Compared with blank control group and solvent control group,both insulin-induced SCF expression reduced after pretreated with LY-294002 or PD-98059 (all P＜0.05).Conclusions Insulin can promote SMCs proliferation,and in certain range the induction of SCF synthesis in colon SMCs is in dose and time dependent manners.The effect may be related to PI3K and MAPK signal pathway.

Objective To investigate the principles of combined traditional Chinese and western medicine treatment of early postoperation inflammatory small bowel obstruction(EPISBO).Methods 93 cases of EPISBO were randomly divided into two groups including group A,47 cases received combined traditional Chinese and west- ern medicine treatment,and group B 46 received only western medicine treatment.Results 93 patients were all cured through nonsurgical treatment.The combined traditional Chinese and western medicine treatment was better in symtom improvement and reduce of expense.Conclusion The principle treatment of EPISBO is mainly nonsurgical. The traditional Chinese medicine and somatostatin are very important.

Objective To investigate the effects of different analgesia protocols on immune function in patients undergoing surgery for ovarian neoplasms after neoadjuvant chemotherapy.Methods Forty ovarian neoplasms patients ageing from 31 to 62 years undergoing neoadjuvant chemotherapy were randomly divided into 2groups:epidural analgesia group (group E,n =20) and intravenous analgesia group (group Ⅰ,n =20).All patients underwent surgery under total intravenous general anesthesia.Patients in group E were given 2 mg morphine in epidural space at half an hour before abdomen was closed,then epidural analgesia pump was installed,with ropivacaine composite morphine in the pump.Analgesia time was 48 h.Patients in group Ⅰ were given sufentanil 0.1 μg/kg intravenously at half an hour before abdomen was closed,then analgesia pump was installed,with sufentanil compound flurbiprofen ester in the pump.Analgesia time was 48 h.The cervical venous blood samples were obtained from the patients at 30 min before surgery(T0),immediately(T1),12 h(T2),24 h(T3) and 48 h (T4) after the end of operation for determination of the expression of CD3 +,CD4 +,CD8 + on T cells and natural killer cell.Visual analogue scales (VAS)and adverse reaction at T2-4 were recorded.Results No statistical difference was found between group E and group Ⅰ in VAS and adverse reaction.Compared with T0,CD3 +,CD4 + T-lymphocytes,CD4 +/CD8 +,and NK cell decreased.Compared with group E,CD8 + increased at T1-2in both groups and at T3-4 in group Ⅰ.CD3 +,CD4+ T-lymphocytes,CD4+/CD8 + and NK cell decreased while CD8 + increased at T2-4 in group Ⅰ.Conclusion Epidural analgesia may be better to promote the immunologic function of ovarian neoplasms patients undergoing surgery after neoadjuvant chemotherapy.

To obtain the key genes for chlorogenic acid biosynthesis of Lonicera hypoglauca, four new genes ware obtained from the our dataset of L. hypoglauca. And we also predicted the structure and function of LHPAL4, LHHCT1 , LHHCT2 and LHHCT3 proteins. The phylogenetic tree showed that LHPAL4 was closely related with LHPAL1, LHHCT1 was closely related with LHHCT3, LHHCT2 clustered into a single group. By Real-time PCR to detect the gene expressed level in different organs of L. hypoglauca, we found that the transcripted level of LHPAL4, LHHCT1 and LHHCT3 was the highest in defeat flowers, and the transcripted level of LHHCT2 was the highest in leaves. These result provided a basis to further analysis the mechanism of active ingredients in different organs, as well as the element for in vitro biosynthesis of active ingredients.
Chlorogenic Acid ; metabolism ; Cloning, Molecular ; Computational Biology ; Gene Expression Regulation, Plant ; Lonicera ; chemistry ; classification ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary

ObjectiveTo determine the levels of type Ⅳ collagen and matrix metalloproteinase-9 ( MMP-9 ) in the serum and kidney of rats with the multiple organ dysfunction syndrome ( MODS ) and investigate the mechanism of extracellular matrix damage in renal failure of MODS.MethodsForty adult male Sprague- Dawley (SD) rats were randomly (ramdom number) divided into two groups,namely the normal control group ( n =8 ) and the MODS model group ( n =32 ).The rats of model group were further divided into four sub-groups as per different intervals,6 h,12 h,24 h and 48 h,after modeling (n =8 in each).The animal models of MODS were established by two hits,the left eyeball of each model rat was removed to bleed to 2 ml bloocd/100 g body weight and lipopolysaccharide ( LPS,5 mg/kg) was injected into intraperitoneal cavity of model rats four hours later. The same volume of saline instead was injected intraperitoneally into rats of control group.All rats were sacrificed at different intervals.Creatinine (Cr)and blood urea nitrogen (BUN) were determined by using a Hitachi Automatic Biochemical Analyzer.The histological changes in renal tissue were observed under light microscope and transmission electronic microscope.The levels of serum type Ⅳ collagen and MMP - 9 protein were measured by using enzyme linked immunosorbent assay (ELISA).Type Ⅳ collagen and MMP- 9 protein levels in renal tissue were detected by western blot.One - way ANOVA was used for comparison among multiple groups.Results Compared with the control group,Cr and BUN were significantly higher in MODS group ( P ＜0.05 ).There were no histopathological changes in kidney of rats in control group,and the renal injury was serious in rats with MODS.The remarkable edema of basement membrane and defect of collagen fibers in renal tissue were observed in MODS group.Compared with the control group,the levels of MMP-9 in serum increased 6-48 hours after modeling and peaking at interval of 12 hours after modeling ( P ＜ 0.05 ).The protein levels of type Ⅳ collagen increased not significantly in 6 h group and 12 h group in comparison with control group (P ＞ 0.05 ),while those in 24 h group and 48 h group significantly increased ( P ＜ 0.01 ).The levels of MMP -9 in renal tissue of rats with MODS increased in 6 h group and 12 h group (P＜0.05),peaked in 24 h group ( P ＜ 0.05 ),and decreased in 48 hours.However,the level of Ⅳ collagen in serum of rats with MODS decreased significantly 6-48 hours after modeling (P ＜ 0.05 ) Conclusions The injury of extracellular matrix is an important factor to the kidney damage in MODS and it may he used for early diagnose and as a treatment target for kidney injury in MODS.

n clinic.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Arrhythmias, Cardiac ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective To investigate the effects of ursolic acid (UA) on cisplatin (CDDP)-induced expres_sion of transient receptor potential vanilloid 1 (TRPV1) in mouse cochlea .Methods Sixty BALB/c mice were ran_domly divided into 4 groups (15 mice in each group) and received introperitoneal injection once daily for 5 days:Control group (normol saline) ,UA group (80 mg/kg/day) ,CDDP group (4 .5 mg/kg/day) ,and CDPP (4 .5 mg/kg/day) plus UA group (80 mg/kg/day) .The expression of TRPV1 in mouse cochlea was determined by immuno_histochemistry ,microscope image analysis and western blot ,and auditory thresholds were evaluated by auditory brainstem response (ABR) measurement .ResuIts The expression of cochlea TRPV1 and ABR threshold shift was significantly increased in the mice treated with CDDP (P< 0 .05) ,as compared with control mice .These effects were prevented by UA treatment (all P<0 .05) .Furthermore ,a linear relationship analysis revealed that the ex_pression of cochlea TRPV1 was significantly correlated with ABR threshold shift(|r|>0 .7 , P<0 .05) .ConcIusion UA effectively attenuated CDDP -induced ototoxicity and improved auditory function through inhibition of TR_PV1 .

PurposeArtifacts of metal implantation severely affect image quality of CT or MRI examination, which restricts the display of local soft tissue lesions after orthopaedic surgery. This study aims to explore the application value of ultrasound examination in local soft tissue lesions after metal implant surgery.Materials and Methods Seventy patients with metal implantation after various orthopedic surgeries received US examination, and such sonographic features were recorded as lesion morphology, size, internal echo, edge, with/without capsule and calcification, and blood flow. Moreover, the clinical and radiologic data of patients like X-ray, CT and MRI were compared with the US results so as to determine the sensitivity of ultrasonic examination.Results The peripheral nerve injury might lead to thickened swelling and echo reduction. Ultrasonography revealed 20 cases of hematoma formation, 5 cases of abscess, 5 cases of pseudoaneurysm, 3 cases of myositis ossificans, 15 cases of bone tumor recurrence and 12 cases of peripheral nerve injury in operative region. Compared with the clinical results , the sensitivity, specificity and accuracy of ultrasound diagnosis of the study reached 89.4%, 75.0% and 88.6%, respectively, which were signiifcantly higher than those of X-ray, CT and MRI (χ2=78.642, 46.596 and 61.371,P<0.01). The ultrasonic results showed that most hematomas had clear boundary with homogenous echo inside to echo-free zone. Abscesses mostly displayed cystic or hybrid echo with floatable intracavitary pus spots echo, and no blood flow signal existed in the central area of color Doppler. Pseudoaneurysm in the US showed cystic mass around artery, blood flow rotating in capsule, and the typical two-stage bi-spectrum signal in the neck of capsule. Myositis ossiifcans showed intramuscular irregular hyperecho with acoustic shadow. When bone tumor recurred, US showed hypoechoic soft tissue mass with moth-eaten bone destruction, and blood flow signals within the mass were often detected.Conclusion Ultrasonography has relatively high accuracy in observing soft tissues around metal implantation, which can be used as a premier method.