Liver fibrosis is an important pathological stage for the progression of chronic liver diseases to liver cirrhosis.Timely and accurate assessment of fibrosis degree plays an important role in guiding the treatment of chronic liver diseases.With the development of molecular biology techniques and wide application of molecular diagnostic techniques,the measurement of novel serum molecular diagnostic markers may contribute to the early diagnosis and precise treatment of liver fibrosis.A number of novel serum molecular markers with a diagnostic value have been identified,including fibroblast factor,matrix metalloproteinases and their inhibitors,extracellular matrix-related markers,and non-coding RNAs.In addition,genomics,proteomics,and metabolomics also have certain diagnostic values.

The human GM CSF cDNAandEGFP cDNA were linked together with IRES and then inserted into the expression vector pCI Egr 1, which was constructed by substituting CMV promoter in pCIneo with the Egr 1 promoter(Egr EG).The vector was transfered into human bone marrow stromal cell line HFCL by Lipofectin TM . The results indicated that the activity of EGFP in transfected cells increased at 18h after exposure to 2.5 Gy. The amounts of secreted GM CSF and CFU GM in serum free supernatants of Egr EG was significantly higher than the control group. EGFP and GM CSF cDNA were successfully integrated and expressed in the cells, which were confirmed by FACS and RT PCR analysis respectively. These in vitro data provide an experimental basis for the in vivo use of gene therapy of GM CSF gene regulated by Egr 1 promoter to protect hematopoiesis from total body irradiation.

Objective To compare the short-term and long-term efficacy of chemotherapy with biotherapy and same chemotherapy followed by biotherapy on malignant melanoma in stage Ⅲ and Ⅳ.Methods 82 cases with malignant melanoma were treated with chemotherapy(dacarbazing,cis-platin and carmustine)or biotherapy(interleukin-2,interferon ? and dendritic cell vaccine)or biochemotherapy(chemotherapy plus biotherapy).Among them,32 cases were received biochemotherapy,26 for biotherapy and 24 for chemotherapy.Therapentic response was assessed every 3 cycles.The median time of follow up was 2 years(1-4 years).Results Response rate was 71.9% for biochemotherapy,46.2% for biotherapy and 54.2% for chemotherapy(P

Objective To explore the chemotherapeutic agents-induced modulating effects of Egr-1 promoter sequence on GM-CSF expression and its protective effect against injury to haematopoiesis due to chemotherapy.Methods Human GM-CSF cDNA and enhanced green fluorescent protein(EGFP) cDNA were linked to internal ribosome entry site(IRES) respectively,and then recombined to pCIneo vector containing Egr-1 promoter(Egr-EG).The vector was transferred into human bone marrow stromal cell line HFCL by lipofection,and the HFCL/EG cells were then finally constructed.MTT assay was performed to determine the effects of cisplatin(DDP),5-fluorouracil(5-FU),gemcitabine(GEM) and paclitaxel(PTX) on the survival rates of HFCL/EG and HFCL cells,and the IC50 of such agents against HFCL/EG and HFCL cells were calculated.The percentage of HFCL/EG cells which positively expressed EGFP was assessed by both flow cytometry and inverted fluorescence microscope.The effects of the active oxygen inhibitor N-acetylcysteine(NAC) on the expression of GM-CSF,which was modulated by promotor Egr-1,were detected by ELISA.The effects of HFCL/EG supernatant on CFU-GM after being exposed to chemotherapeutic agents were examined.Results With different sensitivity to DDP,5-FU,GEM and PTX,HFCL/EG cells were successfully constructed.The drugs showed higher IC50 value against HFCL/EG cells than HFCL cells(P

In order to investigate the feasibility of granulocyte-macrophage colony stimulating factor (GM-CSF) gene therapy, murine GM-CSF cDNA recombinant retrovirus pLXSN/GM was transfered into retrovirus-packaging cell line PA3 17 by electroporation, and the transfected cells were used to infect hematopoietic progenitor cells rich populations. Transfective efficiency of NeoR gene was detected by G418 resistant CFU-GM test, and the results showed 36% them. In the genome of the infected target cells, integrated NeoR gene and GM-CSF cDNA were identified successfully by PCR and Southern Blot analysis respectively. The recombinant plasmids were showed to be capable of expressing GM-CSF mRNA in hematopoietic cells by in situ hybridization. In Dexter culture system, the present of GM-CSF-producing transduced cells inscreased mature nonadherent cell numbers as compared to controls. These results demonstrated that recombinant plasmids were successfully transfected into hematopoietic progenitor cells, and expressed in the cells. Therefore, it provided a basis for further investigation of gene therapy.

Objective:To observe the feasibility of promoting blood circulation by removing blood stasis in target interfering ADUB endometrial hyperplasia.Methods:Endometrial hyperplasia model was established,and the rats were divided into normal group(group A),model group(group B),small,middle and large dosage groups of Chinese drugs(C group,D group,E group,respectively).To observe uterus humid weight,uterus index,morphological change of endometria and expression of VEGF before and after treatment.In addition,30 cases of Anovulatory dysfunctional uterine bleeding were selected and given intrauterine injection of Qumozhibeng Cream twice in the first week after menstrual period,the change of menstrual blood amount were observed after applying medicine for one month,three months and six months.Results:After treatment,the uterus humid weight and uterus index decreased(P

Objective To observe the effects of treadmill training on the recovery of neurological function and the expression of HSP70 and C-MYC in the brains of rats with focal cerebral ischemia. Methods Forty-two male adult Sprague-Dawley rats were randomly divided into a sham group ( n =6), a model group (n =18) and a treadmill exercise group (n=18). Focal cerebral ischemia was induced by right middle cerebral artery occlusion (MCAO) in the model group and exercise group using a modified version of Longa's method. The rats in the treadmill exercise group were given treadmill training 6 d per week for 2 weeks after 24 h of MCAO. By contrast, the rats in the sham group and the model group were reared in standard cages. Before the rats were sacrificed at the 3rd, 7th and 14th d after MCAO, their neurological functions were tested using modified neurological severity scores ( mNSS) , and the mRNA and protein levels of HSP70 and C-MYC were detected using a reverse transcriptase-polymerase chain reaction (RT-PCR) , immunohistochemistry and Western blotting. Results Neurological function in the exercise group at the 7th and 14th days after MCAO had improved significantly compared with the control and model groups.The mRNA and protein levels of HSP70 and C-MYC were significantly upregulated at the 7th and 14th days. Conclusions Treadmill training can improve neurological function by upregulating the expression of HSP70 and C-MYC in the ischemic brain after MCAO.

Migraine is one of the common and frequently encountered diseases. The study proves that 5-hydroxytryptamine (5-HT) receptor, plays an important role in the occurrence of migraine. Rat striatum was used for preparation of the cell membrane stationary phase (CMSP) in our experiments. The cell membrane chromatography (CMC)-offline-HPLC system was applied to specifically recognize the components from the drug pair of Chuanxiong Rhizoma and Angelicae Dahuricae Radix, which interact with the receptors on CMSP. The dissociation equilibrium constant (KD) was measured in a rat striatum/CMC system, performed by continuously pumping sumatriptan, a 5-HT1D agonist, ranging from 2.42 x 10(-8) to 4.84 x 10(-7) mol · L(-1) through a CMC column, and the capacity factors (k') were recorded. The KD value obtained from the model was (4.59 ± 0.33) x 10(-6) mol · L(-1) for imperatorin, and the rat model of migraine induced by nitroglycerin was applied to validate the pharmacological effects of the drug pair. The results indicated that the CMC method could be a quick and efficient way for characterizing the drug-receptor interactions in vitro.
Angelica ; chemistry ; Animals ; Cell Membrane ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; chemistry ; Male ; Migraine Disorders ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT1D ; chemistry ; Serotonin Receptor Agonists ; analysis

OBJECTIVETo investigate the effects of Artesunate(Art) on the LX-2 cell.

METHODSThe cultured hepatic stellate cells were divided into control group and Art-treated groups with 250,350,450 µmol/L. The rate of cellular proliferation was detected by MIT assay, the content of ceramide (Cer)was determined by HPLC method, the content of hydroxyproline (Hyp) was determined by enzyme digestion method, the expressions of PPAR-γ, p53 and Caspase 3 were detected by Western blot.

RESULTSCompared with control group, IX-2 treated with Art were inhibited in a concentration-dependent manner(P < 0.01). Art could significantly increase the content of cerarnide in LX-2 ( P <0.01), and the content of Hyp was significantly decreased (P <0.05, P <0.01). The expressions of PPAR-γ, p53 and Caspase 3 were increased compared with that of control group(P < 0.01).

CONCLUSIONArtesunate could inhibit the proliferation and induce apoptosis of hepatic stellate cells through upregulating ceramide.

Apoptosis ; Artemisinins ; pharmacology ; Caspase 3 ; metabolism ; Cell Line ; Cell Proliferation ; Ceramides ; metabolism ; Hepatic Stellate Cells ; drug effects ; Humans ; Hydroxyproline ; metabolism ; Liver Cirrhosis ; PPAR gamma ; metabolism

Objective To systematically evaluate the effects of lidocaine on female patients during urinary catheterization.Methods A literature search was conducted in following databases as the Cochrane Library,EMbase,PubMed,CBM,CNKI,VIP,and Wanfang data.All included RCTs were screened and assessed according to the standard of Cochrane systematic review.The data were analyzed by RevMan 5.0 software.Results A total of 11 RCTs were included,involving 737 patients in the experimental group,666 patients in the control group.The results of Meta-analysis showed that there were significant differences between the two groups in the pain caused by intubation,one-time success rate of intubation,and agitation rate in anesthesia recovery period.Conclusions Using lidocaine for female patients before catheterization may reduce the pain,increase the one-time success rate of intubation,and reduce agitation rate in anesthesia recovery period.The results still need to be confirmed by more high-quality and large-sample RCTs.