Objective To evaluate the effects of metformin on transforming growth factor-betal (TGF-β 1)-induced epithelial-mesenchymal transition (EMT) in and invasion of the human melanoma cell line 1205Lu.Methods In vitro cultured 1205Lu cells were divided into 3 groups to be treated with serumfree culture medium (blank control group),serum-free culture medium containing 5 ng/ml TGF-β 1 (TGF-β 1 group) and serum-free culture medium containing 5 ng/ml TGF-β 1 and 1 mmol/L metformin (TGF-β1 + metformin group),respectively.After 48-hour treatment,morphological changes of 1205Lu cells in the above groups were observed by using a microscope,and photos were taken at the same time.Transwell invasion assay was performed to estimate cellular invasive activity,Western blot analysis and real-time fluorescence-based quantitative RT-PCR were conducted to determine the mRNA and protein expression of N-cadherin and matrix metalloproteinase-9 (MMP-9),respectively.Statistical analysis was carried out by one-way analysis of variance (ANOVA) and least significant difference (LSD) test.Results Compared with the blank control group,the stimulation of TGF-β 1 could induce the epithelial-mesenchymal transition (EMT)-like morphological changes in 1205Lu cells,while TGF-β 1 combined with metformin could reverse the EMT-like morpho-logical changes.The number of 1205Lu cells crossing the transwell Matrigel per high-power field (× 200) was significantly higher in the TGF-β1 group (412.2 ± 13.427) than in the blank control group (194.1 ± 8.295) and TGF-β1 + metformin group (175.3 ± 8.693).Compared with the blank control group and TGF-β1 + metformin group,the TGF-β1 group also showed significantly increased mRNA and protein expression of N-cadherin (mRNA:6.678 ± 0.043 vs.1.000 ± 0.000,1.035 ± 0.015;protein:1.963 ± 0.016 vs.0.603 ± 0.029,0.207 ± 0.009) and MMP-9 (mRNA:5.351 ± 0.604 vs.1.000 ± 0.000,0.930 ±0.130;protein:1.243 ± 0.027 vs.0.575 ± 0.009,0.629 ± 0.008).Conclusion Metformin can obviously inhibit TGF-β1-induced EMT-like morphological changes in,the capacity to penetrate Matrigel-coated transwell chambers of and the mRNA and protein expression of N-cadherin and MMP-9 in 1205Lu cells.

A molecular template synthetic polymer highly selective for norfloxacinum was prepared by a molecular imprinting technique. The selective binding characteristics of the template polymer was evaluated by Scatchard analysis. The multiple-sites binding model was used to calculate the maximum numher of binding sites and the dissociation constant. The results showed that the template polymer using methacrylic acid (MAA) as functional monomer could form two kinds of binding sites. The dissociation constants were estimated to be Kd1 = 2.9 × 10-5mol/L and Kd2 = 3.2 × 10-3 mol/L. The selective binding experiment for substrates indicated that the polymer gave much higher affinity and selectivity for norfloxacin than for acid pipemidic and cefalexin. It is possible to be a good adsorption and binding material in the selective enrichment and determination of trace norfloxacin in complex biosamples.

The starting material L-tryptophan reacted with 4-chlorobenzaldehyde via Pictet-Spengler condensation and followed by oxidation and decarboxylation to afford the 1-(4-chlorophenyl)-beta-carboline. The intermediate was further reacted with alkyl halogenide by N(9)-alkylation and N2-quaternarization to obtain 12 novel 1-(4-chlorophenyl)-beta-carboline derivatives. The chemical structures of all target compounds were characterized by elemental analyses, MS and 1H NMR spectra. The antitumor activities of the target compounds were evaluated by MTT method. The results demonstrated that N2-quaternarized compounds enhanced the antitumor activity significantly. In particular, compound 15 was found to be the most potent compound with IC50 values lower than 5 micromol x L(-1) against 6 human tumor cells. These results confirmed that the N2-alkyl or aralkyl substituent on the beta-carboline ring played an important role in the modulation of the antitumor activities.

Objective To investigate the changes of thyroid hormones in patients with peripartum car-diomyopathy. Methods Seventy-two patients with peripartum cardiomyopathy were consecutively enrolled and 72 healthy women who had normal delivery were regarded as control. Among all the subjects , free triiodothyronine (FT3), free thyroxine (FT4) and thyroid-stimulating hormone (TSH) in the serum and high sensitive C-reactive protein (hs-CRP), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in the plasma were measured. Results The levels of serum FT3 and FT4 in peripartum cardiomyopathy group were significantly higher than those in control group (P < 0.05). And the level of serum TSH in peripartum cardiomyopathy group was significantly low-er than that in control group (P < 0.05). There were 24 cases with hyperthyroidism in peripartum cardiomyopa-thy group while no patient in the control group had hyperthyroidism (P < 0.05). The levels of plasma hs-CRP and IFN-γ in peripartum cardiomyopathy group were significantly higher than those in the control group (P <0.05). And the level of IL-4 in peripartum cardiomyopathy group was significantly lower than that in the control group (P < 0.05). Conclusions Serum thyroid hormones elevated in patients with peripartum cardiomyopathy and its mechanism might be related to abnormal immune reaction.

Objective To evaluate if expression of serum retinol-binding protein 4 (RBP4) concentrations in cholesterol gallstone disease.Methods Serum RBP4 levels of 100 cholesterol gallstone disease patients (cholesterol gallstone disease group) and 147 healthy controls (healthy control group) were measured by enzyme-linked immunosorbent assay and further correlated with clinical and biochemical characteristics,including insulin resistance and renal function.The chemical composition of gallstones was determined by postoperative pathology.Results The level of RBP4 in cholesterol gallstone disease group was significantly lower than that in healthy control group [(30.57 ± 13.64 ) mg/L vs.(41.52 ± 20.25 ) mg/L](P＜ 0.01 ).The level of RBP4 was also associated with gallstone occurrence (OR =0.93,95％ CI:0.88 -0.96,P =0.004).Serum RBP4 levels of all subjects were positively correlated with total cholesterol,triglyceride,creatinine,insulin resistance and albumin ( P ＜ 0.05 or ＜ 0.01 ),and negatively correlated with aspartate aminotransferase (P ＜0.01).In multivariate analysis,cholesterol gallstone formation was significantly associated with a lower serum RBP4 level (OR =2.97,95％ CI:1.15 -7.68,P =0.025 ).Both gallstone patients and controls were subdivided into two groups according to ceatinine:≥ 88.40 μ mol/L group and ＜ 88.40 μ mol/L group.Patients with gallstones were found to have significantly lower serum RBP4 levels than controls in both subgroups (P =0.012,0.045 ).According to GFR,both gallstone patients and controls were subdivided into ≥ 90 ml/(min· 1.73 m2) group,60 - 89 ml/(min· 1.73 m2) group and ＜60 ml/ (min· 1.73 m2) group.It showed that a lower GFR was associated with greater serum RBP4 level in healthy control group.This trend was not noted in cholesterol gallstone disease group.Conclusions Serum RBP4 decreases in cholesterol gallstone disease independent of renal function.The relationship between liver function and RBP4 level in these patients deserves further investigation.

Objective To clarify whether the regulatory effect of all-trans-retinoic acid (atRA) on the antigen presentation function of dendritic cell(DC) is tightly associated with NF-κB signaling pathway.Also to clarify atRA mainly affected differentiated DC or influence the differentiation procedure from bone marrow cell to DC.Methods MOG35-55 immunized C57BL/6J mice received administration of atRA or not,splenic DC and CD4 cells isolated from immunized mice were subjected to in vitro cross-culture,treated with IL-12 and IL-23 respectively.Th1 and Th17 polarization of stimulated CD4 cells were determined by intracellular staining and FACS analysis,while the production of their corresponding cytokines,IFN-γ and IL-17,were measured by ELISA.Bone marrow cells were isolated from the femurs of the na(i)ve mice,treated with IL-4 and GM-CSF with or without synergistic RA treatment.MHC Ⅱ and CD11c molecules on the DC were assayed by immune staining and FACS analysis,their antigen presentation functions were decided by the proliferation and cytokine production of the Th1 effecter cells stimulated by antigen pulsed DC.To investigate the status of the NF-κB signaling pathway,the amount of phospho-Ser536 NF-κB p65 in the whole DC lysate and total NF-κB p65 in the nucleus were detected by Western blot.Finally,selective RA receptor antagonists were studied to figure out which receptor was majorly involved in the regulatory effect of RA on DC.Results Splenic DC from RA treated mice showed significantly decreased function of presenting antigen to stimulate CD4 cell polarization and cytokine production.Compared with untreated control,RA in vitro treated DC showed decreased expression of MHC Ⅱ and CD11c on its cell surface,which was accompanied with depressed function of stimulating Th1 cell proliferation and IFN-γ production.Further study revealed that RA mainly affect the differentiation procedure from BMC to DC,however,has no significant effect on differentiated DC as the aspects of MHC Ⅱ,CD11c expression,responder cell proliferation and cytokine production were evaluated.Decreased amount of phosphor-Ser536 NF-κB p65 in the whole cell lysate and total NF-κB p65 in the nucleus was investigated in RA treated DC,and decreased antigen presentation function of RA treated DC always came together with diminished activation of NF-κB signaling pathway.Finally it was demonstrated that RARβγ antagonist,but not RARα antagonist,could entirely block the RA effect on DC.Conclusion Retinoic acid inhibit the differentiation of DC as well as decrease its antigen presentation function,which might be resulted from the inactivation of NF-κB p65 signaling pathway and mediated by RARβγ.

Humans ; Medicine ; Medicine, Chinese Traditional ; methods

Objective:To evaluate the relationship between breast reconstruction and postoperative complications by meta-analysis.Methods:Through a defined search strategy, related literature was collected in databases from PubMed, MEDLINE, EMBASE, Cochrane, CNKI, Wanfang Database and VIP Database, from January 1990 to November 2017. Data were extracted and each merged data was analyzed using RevMan 5.3 software. The postoperative complications between transverse rectus abdominis musculocutaneous-flap (TRAM) and deep inferior epigastric perforatorflap (DIEP), TRAM and latissimus dorsiflap (LDF), LDF and LDF+ prosthesis, LDF+ prosthesis and simple prosthesis implantation were compared.Results:Twenty-nine papers met inclusion criteria of our study. The Meta-analysis results showed that the risk of local flap necrosis, seroma, infection, fat liquefaction, abdominal wall hernia and abdominal bulging in TRAM group were higher than those in DIEP group, and the differences were significant. There was no significant difference in total flap necrosis, wound dehiscence, venous congestion between the two groups; the risk of total flap necrosis, wound dehiscence and infection in TRAM group were higher than those in LDF group, and the differences were significant. The risk of seroma in TRAM group was lower than that in LDF group, and the differences were significant. There was no significant difference in fat liquefaction between the two groups; there was no significant difference in the postoperative complications between LDF group and LDF+ prosthesis group; the risk of seroma in LDF+ prosthetic group was higher than that in prosthetic group, and the differences was significant. The risk of prosthetic capsular contracture and prosthesis displacement in LDF+ prosthetic group were lower than those in prosthetic group, and the differences were significant. There was no significant difference in prosthesis exposure and infection between the two groups.Conclusions:DIEP has most of the advantages of TRAM and fewer complications. It plays an important role in breast reconstruction in the future. We should make a choice of breast reconstruction methods according to the patient＇s conditions as far as possible in clinical practice.

Objective To explore the mechanism of percutaneous laser disc decompression (PLDD) for treating prolapse of intervertebral disc. Methods 15 rabbits were divided into 3 groups randomly: sham group, model group with PLDD treatment, model group without PLDD treatment. The nerve conduction velocity (NCV) of L6 nerve root and the activity of phospholipase A2 (PLA2) in the intervertebral disc of L5-6 were determined 2 weeks after the initial surgery. Results NCV in the group with PLDD was significantly faster than that in the group without PLDD (P<0.001); NCV in the group without PLDD was significantly lower than that in the sham group (P<0.001). The activity of PLA2 in the group with PLDD was significantly lower than that in the group without PLDD (P<0.001); The activity of PLA2 in the group without PLDD was significantly higher than that in the sham group (P<0.001). Conclusion The activity of PLA2 in the herniated discs is higher than that in normal discs, which result in NCV falls remarkably. The PLDD can reduces chemical factors such as PLA2.

Objective:To provide a potential platform for transferring specific antigen against fish bacterial diseases based on attenuated Lm (EGDe-ΔactA/ΔinlB).Methods: Attenuated Lm (EGDe-ΔactA/ΔinlB) was used to express outer membrane protein K (Ompk),a conserved and effective vaccine candidate in vibrio.The identification of recombinant strains and detection of antigen genes were operated with PCR and RT-PCR,respectively.Results: The results of PCR showed that Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.The identity of foreign gene was 100％ compared with sequence of NCBI.The analysis of transcription showed that the expressions of Ompk in L-O,L-L-O and L-P-O were significant (P<0.001).Moreover,the expression of Ompk in the condition of antibiotic was higher than that in the BHI without antibiotic (P<0.05).Conclusion: Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.