Objective To study the effect of transient intensive insulin treatment on the serum free fatty acid (FFA) in newly diagnosed type 2 diabetic patients.Methods Sixty-four newly diagnosed type 2 diabetic patients were treated with transient intensive insulin.The fasting plasma glucose (FPG),2 hours post-prandial glucose (2hPG),lipid,fastin insulin (FINS),and serum FFA was examined hefore and after treatment.Results The levels of FPG,2hPG,total cholesterol (TC),triglycerides (TG),low density lipoproteins cholesterol (LDL-C),FFA and HOMA-IR after treatment were (9.68 ± 2.02) mmol/L,(12.77 ± 1.35) mmol/L,(4.26 ± 1.07) mmol/L,(1.52 ± 0.58) mmol/L,(2.50 ±0.75) mmol/L,(435.84 ± 190.94) μmol/L,0.51 ± 0.62,and they decreased obviously compared with those before treatment [(14.66 ± 3.50) mmol/L,(17.43 ±4.89) mmol/L,(5.03 ±0.94) mmol/L,(2.05 ± 1.42) mmol/L,(2.91 ±0.78) mmol/L,(586.68 ±229.45)μmol/L,0.65 ± 0.89](P＜0.05).The level of HOMA-β increased obviously (2.70 ± 0.83 vs.1.74 ± 1.04)(P＜0.05).The increase of HOMA-β and the decrease of HOMA-IR was positively correlated with the decrease of FFA.Conclusion The transient intensive insulin treatment can evidently decrease the level of FFA that can improve beta-cell function and relieve insulin resistance in newly diagnosed type 2 diabetic patients.

Objective To study the effects of low dose sodium arsenite to human bone marrow mesenchymal stem cells(hBMSCs) differentiation during establishing of arsenic-resistant cell model. Methods hBMSCs were prepared in conventional method and continuously exposed to 1μmol/L sodium arsenite for ≥12weeks inv vitro. Forty-eight hours acute arsenite toxicity test was drived to assay if the cells acquired arsenic-resistance. The proliferation capacity of CAsE-hBMSCs was observed by the rate of colony formation.The expression of Oct-4 in CAsE-hBMSCs was assayed by RT-PCR and immunocytochemistry. The expression of ABCG2 in CAsE-hBMSCs was analyzed by RT-PCR. Results hBMSCs continuously exposed to 1μmol/L sodium asenite for ≥12 weeks exhibited dramatic resistance to acute arsenite toxicity. The LC 50 for acute arsenite exposure in CAsE-hBMSCs was 35.59μmol/L versus 18.04μmol/L in control cells. Compared to control cells, the CAsE-hBMSCs didn't show malignant proliferation ability. Expression of Oct-4 gene was positive in 4th, 18th passage hBMSCs and the hBMSCs induced by arsenite for 4 weeks but negative in CAsE-hBMSCs. The expression of Oct-4 protein was positive and weakly positive in 4th passage hBMSCs and CAsE-hBMSCs respectively, and the positive granules of Oct-4 distributed in cytoplasm. The expression of ABCG2 gene in CAsE-hBMSCs was obviously lower than that in control cells ( P <0.001). Conclusion Human bone marrow mesenchymal stem cells could be induced to differentiation by low dose sodium arsenite.

ObjectiveTo investigate the expressions of nerve growth factor (NGF) and its receptors TrkA and p75NTR in dermatofibrosarcoma protuberans and dermatofibroma.MethodsAvidin-biotin immunohistochemical(ABC) method was used to detect the expressions of NGF and its receptors TrkA and p75NTR in paraffin-embedded tissue specimens from 17 cases of DFSP and 15 cases of dermatofibroma.Results NGF and TrkA were highly expressed in both DFSP and dermatofibroma specimens,with no significant difference between the two groups of specimens (x2 =0.11,0.02,respectively,both P ＞ 0.05),while the expression of p75NTR was significantly higher in DFSP than in dermatofibroma specimens(x2 =32,P ＜ 0.01 ).The expression of NGF was positively correlated with that of p75NTR in DFSP(R2 =0.623,P ＜ 0.01 ).ConclusionNGF may play a certain role in the development of DFSP via its high-affinity receptor TrkA and low-affinity receptor p75NTR.

Objective To detect the expressions of nerve growth factor (NGF) and its receptors tyrosine kinase A (TrkA) as well as p75 neurotrophin receptor (p75NTR) in the lesions of lichen planus.Methods Biopsy specimens were collected from the lesions of 32 patients with lichen planus and normal skin of 12 healthy human controls and subjected to paraffin embedding.Immunohistochemical avidin-biotin complex (ABC) method was used to detect the expressions of NGF,TrkA and p75NTR.Results NGF and TrkA,which were located in the cytoplasm of keratinocytes,were strongly or moderately expressed in the lesional skin specimens,but absent or weakly expressed in the normal skin specimens (both P ＜ 0.01).No significant differences were observed in the expression of p75NTR between the lesional and normal skin specimens,or in the expressions of NGF,TrkA or p75NTR among specimens from patients in different age groups,patients of different gender or lesions at different sites (all P ＞ 0.05).There was a positive correlation between the expression of NGF and TrkA in the lesions of lichen planus (R2 =0.535,P ＜ 0.01).Conclusion NGF may play a certain role in the development of lichen planus via its highaffinity receptor TrkA.

AIM: To investigate the pharmacological effects and mechanism of ethyl ferulate (EF). METHODS: The platelet congregate rate was observed by congregater TYXN-91 and the platelet intracellular calcium oscillation was observed by laser scanning. The acute liver injury model of mice was made by using the CCl 4 156 mg?kg -1, ip. Then the levels of ALT and AST were determined in serum and the levels of MDA and SOD in the liver. RESULTS: The inhibition rate of the platelet congregate were 26.3%? 3.3%, 33.4%? 2.4%, 73.4%? 3.1%, and 94.9%? 2.7% (n=8) in different concentration( 0.1, 0.5, 1.5, and 3.0 mmol?L -1)of ethyl ferulate,respectively. It was higher than those in the same concentrations of ferulic acid. The change of the fluctuation of calcium (?FI 4.6? 1.7) in EF group was much lower than the rest level ( 10.3? 2.6) (n=8,P

OBJECTIVE:To evaluate the therapeutic effect of compound glycyrrhizin plus mizolastine for patients with generalized neurodermatitis(GND)and its influence on patietents' psychology.METHODS:60 patients with GND were randomly assigned to receive compound glycyrrhizin plus mizolastine(treatment group)or mizolasitine alone(control group)for 28 days.The psychological factors were examined before and after treatment using self-rating anxiety scale(SAS),and the curative effects and the psychological changes between two groups were compared.RESULTS:The SAS scores of treatment group and control group were 30.53? 1.50 and 33.20? 1.67,respectively(P

Objective To investigate the potential role of Spa-1 in the development and metastasis of EMPD.Methods Tissue specimens were resected from 17 patients with primary EMPD,12 patients with invasive EMPD and 9 normal human controls.Immunohistochemistry was performed to measure the expression of spa-1 and Ki-67.Results Positive staining was observed for Spa-1 in cytoplasm,and for Ki-67 in cell nuclei.Spa-1 and Ki-67 were weakly expressed in the basal layer of normal skin.The expression of Spa-1 and Ki-67 in EMPD tissue were statistically higher than those in the normal control tissue (both P<0.01).Increased expression of Spa-1 was noted in invasive EMPD tissue compared with in situ EMPD tissue.The expression of Spa-1 was positively correlated with that of Ki-67 in the tissue of EMPD.Conclusion Spa-1 may play a certain role in the initiation and progression of EMPD.

Objective To investigate the inhibitory effects of L. monocytogenes expressing melanomainhibiting activity(MIA)gene on malignant melanoma.Methods The plasmid pERL3-hly-MIA was constructed and used to transform live L.monocytogenes by electroporation.Atier culture.the transformed L.monocytogenes,namely LM-hly-MIA,was harvested for the detection of MIA protein by Westem Blot.A total of 24 mice were divided into three groups,namely control group,LM group and LM-hly-MIA group,to be immunized with physiological saline,L.monocytogenes and LM-hly-MIA,respectively.One day after the immunization.mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentration of 1×107/mL.One week later.reimmunization was performed.The size and weight of tumors were observed and measured in these mice.Results Enzyme digestion and Western blot confirmed the Successful construction of plasmid pERL3-hly-MIA.The average weight of tumors in mice was 4.33±0.91 gram.3.36 ±0.41 gram and 1.89±0.52 gram,respectively in the control group,LM group and LM-hly-MIA group,respectively(P＜0.05).with the inhibition rates of tumor growth being 22.4%and 61.5%in LM group and LM-hly-MIA group.respectively.Conclusion The attenuated strain of L.monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.