Background:Collagen triple helix repeat containing protein-1(Cthrc1)has been reported playing an important role in liver diseases,especially in liver fibrosis,however,its effect on hepatic stellate cells proliferation is not fully clear. Aims:To investigate the effect of Cthrc1 on proliferation of hepatic stellate cells. Methods:Recombinant adenovirus vector of Cthrc1 was constructed. After injecting Ad-Cthrc1 through tail vein,mRNA and protein expressions of Cthrc1 were determined by real-time PCR and Western blotting,respectively. Liver fibrosis model was established by bile duct ligation and fed with 3,5-diethoxycarbonyl-1,4-dihydrocollidine(DDC)in mice,respectively. The liver fibrosis mice were injected with Ad-Cthrc1 or Ad-GFP through tail vein. Immunofluorescence was used to determine number of hepatic stellate cells. Results:Recombinant Cthrc1-adenovirus vector was successfully constructed. Real-time PCR and Western blotting showed that mRNA and protein expressions of Cthrc1 were increased in Ad-Cthrc1 group than in control group. HE and Masson staining demonstrated that mice model of liver fibrosis was successfully established. Immunofluorescence showed that overexpression of Cthrc1 inhibited significantly the proliferation of hepatic stellate cells. Conclusions:Recombinant adenovirus vector of Ad-Cthrc1 constructed can express stably in vivo,and inhibit the proliferation of hepatic stellate cells. Therefore,Cthrc1 may become a potential target for treatment of liver fibrosis.

Objective To explore the effect of osthole on mast cells and expression of STAT5 gene and protein in them. Methods Passive cutaneous anaphylaxis was made in mice to copy eczema model, then the allergic mast cells were separated, and the ovalbumin was used to induce allergy of mast cells. Different concentrations of osthole were used to intervene the sensitized mast cells.Then the sensitized mast cells were divided into blank control group, osthole high-dose group and low-dose group.At the end of the experiment, morphology of the mast cells was detected by immunofluorescence technology.MTT assay was used to detect the influence of drugs on mast cells proliferation. RT-PCR and Western blotting were used to detect the expression of STAT5 gene and protein. Results As compared with blank control group, the number of mast cells in the osthole groups was significantly reduced, cells and nuclei obviously shrank, even apoptosis of some cells could be observed; the inhibition rate of mast cells in osthole groups was significantly increased in concentration-dependent manner ( P<0. 01 ) . As compared with blank control group (2.16±0.57), gene expression of STAT5 was significantly decreased in osthole high-dose group (0.59±0.12) and low-dose group (0.82±0.13) (P<0.01).The protein expression of STAT5 was also significantly decreased in osthole high-dose group and low-dose group as compared with blank control group (P<0.01). Conclusion Osthole can inhibit the proliferation of sensitized mast cells, and reduce the expression of STAT5 gene and protein.

Objective To study the efficiacy and safety of transfacet approach decompression to treat thoracic spinal stenosis caused by anterior compression.Methods Thirty-three patients with thoracic spinal stenosis caused by anterior compression were treated in our institution from April 2005 to April 2009.Nineteen patients with more than 12 months follow-up were included in this study.Among of them,10 were male and 9 were female,with the age ranged from 33 to 77 years(mean,55.9 years).The causes of compression for spinal stenosis included ossification of posterior longitudinal ligament(OPLL)in 5 cases,thoracic disc herniation(TDH)in 11 cases,OPLL with ossification of ligamentum flavum(OLF)in 2 cases and TDH with OLF in 1 case.All patients underwent anterior decompression via a transfacet approach combined with anterior fusion and posterior fixation.The modified Japanese Orthopaedic Association(JOA)score system and Nurick Myelopathy grade were used to evaluate the outcomes.Results The operation time ranged from 180 to 480 min,with an average of 299.5 min and the blood loss was varied from 250 to 2200 ml,with an average of 918.5 ml.Among 7 cases with OPLL(including combined with OLF),2 patients developed neurologic deterioration and 1 patient developed cerebrospinal fluid leakage.There were no neurologic deterioration,cerebrospinal fluid leakage and other complications occurred in 12 cases with TDH(including combined with OLF).The follow-up ranged from 12 to 54 months(mean,28.6 months).The preoperative JOA score ranged from 2 to 11(mean,6.3).The JOA score in the last follow-up ranged from 5 to 11(mean,8.6).According to Nurick Myelopathy grade,the preoperative grade was 0 in 2 cases,1 in 2,2 in 4,3 in 5,4 in 2,and 5 in 4.The number of postoperative grade was 6,6,3,3,1 and 0 respectively.Conclusion Satisfactory decompression could be achieved by using transfacet approach for thoracic spinal stenosis caused by anterior compression.The approach is a safe and promising alternative for thoracic spinal anterior decompression.

OBJECTIVE: To explore whether pre-administration of Ganoderma lucidum spore (GASP) can reduce incidence of neural tube defects (NTDs) induced by all-trans retinoic acid (ATRA) in pregnant mice. METHODS: Twenty pregnant mice were randomly divided into four groups: normal control group, solvent-treated group, ATRA-induced group, and GASP-treated plus ATRA-induced group. GASP solution, which was prepared with solvent (sodium carboxymethyl cellulose), was fed to the pregnant mice in the GASP-treated plus ATRA-induced group twice a day from embryo (E) 0 d to E10.5 d. The same dose of solvent was given to the pregnant mice in the solvent-treated group. At E7.75 d, ATRA (50 mg/kg) was given to the pregnant mice in both ATRA-induced group and GASP-treated plus ATRA-induced group for single time. Embryos were sampled from pregnant mice at E10.5 d. Then the incidence rate of NTDs in mouse embryo was calculated and the crown-rump length of mouse embryo was measured. The positive rate of nestin expression and the distribution of cell cycle of embryonic neural tube neuroepithelial cells were detected by histochemical staining technique and flow cytometry respectively. Reverse transcription-polymerase chain reaction method was used to detect the gene expressions of cyclin-dependent protein kinase 2 (Cdk2) and Cdk4 mRNAs. RESULTS: The incidence rates of NTDs in mouse embryos in the ATRA-induced group and the GASP-treated plus ATRA-induced group were 79.41% and 21.67% respectively, while the crown-rump length of mouse embryos in these two groups were (3.62+/-1.27) mm and (5.84+/-0.92) mm respectively. The positive rate of nestin expression in embryonic neural tube neuroepithelial cells of mouse embryo at E10.5 d in the ATRA-induced group was 32.44%, while that in the GASP-treated plus ATRA-induced group was 77.65%. The cell cycle of embryonic neural tube neuroepithelial cells was obviously arrested at G(0)/G(1) phase in the ATRA-induced group as compared with that in the GASP-treated plus ATRA-induced group. The Cdk4 mRNA was transcripted at a high level in embryonic neural tube in the GASP-treated plus ATRA-induced group, but the Cdk2 mRNA was not detected in this group. CONCLUSION: Pre-administration of GASP can reduce the incidence of NTDs induced by ATRA in pregnant mice.

Adolescent ; Adult ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; Poisoning ; epidemiology ; Young Adult

OBJECTIVETo control the quality of the product, quantitative fingerprint was used to evaluate the composition of the amino acids in the Xingnao Tongluo injection.

METHODThe method of the quantitative fingerprint to the amino acids composition was established through AccQ Tag precolumn derivatization. The quality was evaluated by the quantitative test of the amino acids and the similarity in ten batches.

RESULTThe Xingnao Tongluo injection contained 12 amino acids and the contents of these amino acids were stable. All the ten batches of the samples had similarity of more than 0.90.

CONCLUSIONThe method was accurate, feasible and could be a simple and effective way to evaluate the quality of the traditional Chinese medicine.

Some studies indicate that adipose derived stem cells (ADSCs) can differentiate into adipogenic, chondrogenic, myogenic, and osteogenic cells in vitro. However, whether ADSCs can be induced to differentiate into neural cells in vitro has not been clearly demonstrated. In this study, the ADSCs isolated from the murine adipose tissue were cultured and transfected with the EGFP gene, and then the cells were induced for neural differentiation. The morphology of those ADSCs began to change within two days which developed into characteristics of round cell bodies with several branching extensions, concomitantly expressing EGFP fluorescence. Approximately 60% of the total cell populations were bipolar or multipolar in shape. Some of them appeared to make contact with their neighboring cells. RT-PCR, Western blot and Immunocytochemistry revealed that the expression levels of the markers of neurons and oligodendrocytes such as MAP2, NF-70, Neu N and RIP upon neural induction were increased, but the expression of the special marker of astrocytes, GFAP, was undetectable until 96 h after induction when a small signal was observed. It was concluded that the ADSCs transfected with EGFP possessed the ability to undergo morphologic and phenotypic changes consistent with neural differentiation in vitro. It suggests that these cells might provide an ideal source for further stem cell research with possible therapeutic application for spinal cord injury.

The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 mug/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicity in vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

The formation of osteolytic bone lesions is a key process for osteolytic cancer to metastasize to the bone and is under the control of a set of transcription factors. Recently, the inhibitor of differentiation 1 (Id1) has been linked with angiogenesis, tumorigenesis, metastasis and bone formation. However, the function of Id1 during the process of bone destruction caused by cancer in vivo has not yet been elucidated. We, therefore, examined whether and how Id1 affects the ability of cancer to form osteolytic lesion in vivo. The study used a lentiviral vector overexpressing short hairpin RNA (shRNA) targeting Id1 gene. PC3 cells, a prostate cancer cell line, were transduced with Id1 shRNA or negative control (NC) shRNA before implantation in BALB/c mice. Cells were implanted in a tibial injection model. Tumor formation in bone was monitored by X-ray. The relationship between parathyroid hormone-related protein (PTHrP), an osteolytic factor, and Id1 was analyzed by using immunohistochemistry in tissue sections from osteolytic lesion of the BALB/c mice. Our results showed that Id1 shRNA delivery to PC3 cells by lentivirus caused efficient and stable Id1 gene silencing. In the intratibial model, PC3 cells produced primarily osteolytic lesions in the bone. Eleven of 14 mice in Id1 shRNA group but only 4 of 14 mice in the NC shRNA group developed osteolytic lesions with cortical destruction at 4th week. Mice treated with Id1 shRNA had larger tumor volume in the bone and larger cortical destruction. The expression of PTHrP protein in PC3 cells was not affected by Id1 knockdown in vivo. These results indicate that Id1 may down-regulate the ability of PC3 cells to form osteolytic lesions in vivo and the signal pathway needs to be further investigated.

Objective To compare the effectiveness and safety of different intracranial stents assisted coil embolization of intracranial aneurysms and to discuss the selection of different stent assisted embolization of intracranial aneurysm.Methods From 2007 April to 2012 April,118 cases (a total of 128 wide-neck aneurysms) with intracranial aneurysms were analyzed retrospectively.This included the use of 70 neuroform,38 Enterprise,and 20 Solitaire AB stents forthe treatment of intracranial aneurysms.The successful use,aneurysm occlusion at the immediate post-operation,and early period of peri-operative complications were recorded from those clinical data in order to assess the effectiveness and safety of the different intracranial stents,which assisted coil embolization of intracranial aneurysms.Rank sum test and x2 test were used for statistics.Results Three aneurysms assisted with Neuroform stent were planted unsuccessfully,and the Enterprise and Solitaire stents were placed successfully.The embolism results of three stents after immediate postoperative angiography aneurysm: Neuroform stent occlusion rate was 40.0％ (28/ 70),the tumor residual rate was 38.6 ％ (27/70),and the partial embolization rate was 21.4 ％ (15/70) ; The Enterprise stent occlusion rate was 42.1％ (16/38),the tumor residual rate was 36.8 ％ (14/38),and the partial embolization rate was 21.1％ (8/38).The Solitaire AB stent occlusion rate was 40.0 ％ (8/20),the tumor residual rate was 35.0 ％ (7/20),and the partial embolization rate was 25.0 ％ (5/20).There were not significant differences in aneurismal occlusion (H =0.12,P ＞ 0.05).Early peri-operative complications results were: Neuroform stent occurred in 7,Enterprise frame in 4,and Solitaire AB stent occurred in 2.There were no significant differences in the incidence of complications in the early period after coiling (x2 =0,P ＞ 0.05).Conclusions Three kinds of intracranial stents assisted embolization of intracranial aneurysms are applied safely and effectively.The stent may be chosen according to morphology of parent artery and stent biological character.