Aim To explore the effect and mechanism of resveratrol (Res) on proliferation and differentiation of murine 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were cultured and treated with resveratrol in different dosages.Cell proliferation was analyzed by WST-1 method. Oil red O staining method and spectrophotography were applied to analyze the degree of differentiation. Real-time PCR was applied to detect the mRNA expression of adiponectin and leptin. Western blot was applied to detect the expression of silent information regulator 1 (Sirt1),peroxisome proliferator activated receptor γ (PPARγ) and CCTTA enhancer binding proteinα (C/EBPα).Results Res inhibited proliferation of murine 3T3-L1 preadipocytes in a time-dose dependent manner.The expression levels of adiponectin and leptin mRNA were decreased, and Res also inhibited 3T3-L1 preadipocytes to differentiate into mature adipocytes. Res increased the expression levels of Sirt1 and decreased the expression levels of PPARγ and C/EBPα.Conclusions Resveratrol can inhibit the proliferation and differentiation of 3T3-L1 preadipocytes.The underlying mechanisms may include enhancing expression of Sirt1 and inhibiting expression of PPARγ,C/EBPα which are related to cell differentiation.

Superoxide dismutase(SOD) was purified from Tegillarca granosa and the effects of tributyltin chloride(TBTCl) on the enzyme activity of the SOD were analyzed at different conditios. The results showed that the effects of TBTCl on SOD activity were more ohvious at room temperature than at four degrees centigrade. 58. 1 % of the enzyme activity was reserved after SOD was incubated with 200?g?mL-1 of TBTC1 for 72 hours at room temperature, while 90. 9% of the enzyme activity was reserved at four degrees centigrade. The enzyme activity of SOD was reduced with the increase of TBTC1 concentration and interaction time. TBTCl could reduce heat-resisting, acid-resisting and alkali-resisting abilities of SOD. With TBTCl concentration increase,SOD isozyme bands in the graph of polyacrylamide gel electrophoresis were getting weaker.

BACKGROUND:The interaction of neural network and motor function in post-stroke brain tissue remains unclear.
OBJECTIVE:To observe neural network impairment fol owing subacute stroke by using diffusion tensor imaging, and to investigate the relationship with neurological defects and motor dysfunction.
METHODS:A total of 19 patients after subactue stroke and 20 healthy adults were examined with diffusion tensor imaging. The fol owing parameters were compared:fractional anisotropy, apparent diffusion coefficient, asymmetry indices of fractional anisotropy and apparent diffusion coefficient. The neurological defect and motor function were evaluated with the corresponding scales. The 10-meter walking speed was measured. The correlation of diffusion tensor imaging parameters with the scale scores and 10-meter walking speed was analyzed.
RESULTS AND CONCLUSION:The stroke group exhibited reduced fractional anisotropy value asymmetry and fractional anisotropy value in bilateral posterior limbs of the internal capsule. Apparent diffusion coefficient value asymmetry and apparent diffusion coefficient value in the posterior limb of the internal capsule were lower than control unaffected side (P<0.05). Apparent diffusion coefficient value and apparent diffusion coefficient value asymmetry in posterior limb of the internal capsule showed a strong negative correlation with Fugl-Meyer assessment scores of the lower extremities (P<0.05). Diffusion tensor imaging parameters is closely linked with motor dysfunction of the lower extremities in subacute stroke patients. Local stroke lesion-caused neurological defect is the leading cause of motor dysfunction of the lower extremities.

Glioma is the most common form of brain cancer. Despite recent advances in the treatment of solid tumors, there are few effective treatments for malignant gliomas due to its infiltrative nature. It has important significance to improve the treatment of glioma through in-depth understanding the intracerebral metabolic characteristics and pharmacokinetics of chemotherapeutics. Brain microdialysis (B-MD), an effective method to monitor central nervous system anticancer drug disposition, conditions of drugs through the blood-brain barrier, basic pathophysiologic metabolism, bioactive compounds and the changes of neurotransmitter in brain, provides the unique opportunity to allow the simultaneous determination of unbound concentrations of drugs in several tissues, and directly measure gliomas biochemistry continuously. B-MD has been able to monitor the change of brain drugs, metabolites and neurotransmitters, dynamic analysis of the drug concentration and pharmacological effect after administration, pharmacodynamic interaction between drugs, receptor mechanism of drug transport, as well as feedback information of internal environment. B-MD is expected to provide reference for clinical individual chemotherapy of glioma, but also provide powerful tools for the evaluation of new anticancer drugs in vivo. In this review, a comprehensive overview of B-MD for studies on glioma is elucidated with special emphasis on its application to neurochemistry and pharmacokinetic studies.
Animals ; Antineoplastic Agents ; pharmacokinetics ; Blood-Brain Barrier ; Brain Neoplasms ; metabolism ; Glioma ; metabolism ; Humans ; Magnetic Resonance Spectroscopy ; Metabolomics ; methods ; Microdialysis ; methods ; Neurotransmitter Agents ; pharmacokinetics ; Pharmaceutical Preparations ; metabolism ; Positron-Emission Tomography

Humanized monoclonal antibodies(mAbs) are increasingly widely used in targeted therapy for cancer and some other major diseases.Complementarity-determining region(CDR) grafting makes quantities of humanized mAbs available.Herein,we provide an overview on the strategy and progress of CDR grafting.

Objective To investigate the safety and obtain parameters of microeoagulation of dorsal root entry zone (DREZ) of cervical cord with bipolar foreps on animal model,and provide histological base for clinical application of treatment of brachial plexus avulsiol pain using microcoagulation of dorsal root entry zone.Methods On the base of swine's weight and spinal cord size in similar to human being,it was chosen to be experimental animal.The right DREZs of cervical cord were microcoagulated with bipolar forceps.The swines were fed in normal way.Their activities were observed.The mass change of the cervical cord segment were observed after 3 weeks and the cervical cord segment was fixed with 10% fromalin,paraffin sliced,HE dying.Coagulating space,depth and width were measured under microsope.The coagulating parameter were adjusted according to measuring outcome in order to achieving a most avaliable parameter.Results All post-op swine survived.When the microcogulation were made with bipolar forceps adopted following parame- ters:The distance of between the polar was 2.0 mm;The diameter of polar was 0.3 mm.The inserting depth 2 mm,the coagulated power 18 watt,the coagulated time was 2 second,then the width of lesions of DREZ in cross section was 1.15 mm and the depth of lesions was 3.10 mm,which was consistent with the area of hu- man DREZ of cervical cord.Conclusion The experiment on swine suggested,microcoagulation of DREZ by bipolar forceps is safe and no mortal complications when the testified parameters are adoped,and can achieve the area of DREZ of cervical cord in human.

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

Objective To evaluate the long-term efficacy of vascularized pisiform transfer for patients with Kienb(o)ck's disease in Lichtman stages Ⅲ-Ⅳ. Methods Eleven patients were reviewed to analyze results after lunate resection and vascularized pisiform transfer for Lichtman stages Ⅲ and Ⅳ. There were six men and five women. Age ranged from 20 to 67 years with a average of 41.0±14.3 years. According to Lichtman stage. There were 4 cases in stage Ⅲa, 5 cases in stage Ⅲb, and 2 cases in stage Ⅳ. Assessment criteria included subjective assessment of pain, visual analogue scale (VAS), range of motion (ROM), grip power,Cooney wrist score and radiographic changes on each follow-up visit. The radiographic changes including pis iform bone location, shape, sclerosis change, osteoarthritis, carpal height ratio, Nattrass index, Radioscaphoid angle and ulnar variance were recorded. Results The follow-up periods of all of cases were 61-202 months,with an average of 104.1 months. Pain had improved in 10 patients and disappeared in 7 cases. The VAS score was 2.2±1.9 at follow-up visit. Range of motion of injured wristw as only 65.3% of opposite side. Grip power was 84.3% of the contralateral hand. According to Cooney score, the results were excellent in 1 case, good in 7cases, fair in 2 cases and poor in 1 case, with the excellent and good rate of 72.7%. Radiologically, 8 cases had normal position of the pisiform bone, 2 had volar displacement and 1 had ulnar displacement which leaded to widen scaphopisiform space. Six pisiform bones had normal trabecular structure, three had degenerative changes. Bone sclerosis was seen in 2 cases and osteoarthritis was found in 3 patients. Compared with radiographic parameter before surgery, carpal height ratio and Nattrass index significantly lowered and radioscaphoid angle significantly increased. Conclusion Lunate resection and vascularized pisiform transfer is an effective method for Kienb(o)k′s disease in stages Ⅲ-Ⅳ. Although carpal collapse appeared postoperatively,the results show high patient satisfaction and good function after vascularized bone transplantation.

OBJECTIVETo observe the therapeutic effect of Kangyi Zhongyu Decoction (KZD) combined with gonadotropin releasing hormone-a (GnRH-a) on infertile patients with severe endometriosis.

METHODSSeventy-five infertile patients with the diagnosis of endometriosis confirmed by laparoscope who were scheduled to receive in vitro fertilization and embryo transfer (IVF-ET) were randomly assigned to three groups, they were treated respectively with KZD (A), GnRH-a (B) alone and combined of both (C), and IVF-ET were applied in the patients after 3 months of treatment. The clinical efficacy and adverse reactions in the three groups were observed and the changes of serum cancer antigen 125 (CA125) and endometrial antibody (EMAb) levels before and after treatment were tested.

RESULTSScore of dyspareunia in Group A and C was obvioushy lower than that in Group B after treatment (P <0.01). Pregnancy rate in Group C was higher than that in Group A and B (P <0.05), with the adverse reactions less than in Group B (P <0.01). The positive rate of plasma EMAb was reduced obviously after treatment in Group C with the level lower than that in the other two groups (P<0.05).

CONCLUSIONThe combined use of KZD and GnRH-a is a new method in treating infertile patients with severe endometriosis with ideal effectiveness and fewer adverse reactions, and it could advance the successful rate of reproductive assistant technique.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Embryo Transfer ; Endometriosis ; complications ; drug therapy ; Female ; Fertilization in Vitro ; Gonadotropin-Releasing Hormone ; therapeutic use ; Humans ; Infertility, Female ; drug therapy ; etiology ; Integrative Medicine ; methods ; Phytotherapy ; Treatment Outcome ; Young Adult