OBJECTIVETo analyze the relation of plasma D-dimer levels and incidence of deep venous thrombosis after spinal surgery.

METHODSThe clinical data of 63 patients underwent spinal surgery from October 2009 to October 2010 were retrospective analyzed. There were 40 males and 23 females with an average age of 48 years old(21 to 76) in operation. Operation levels of 15 cases were in cervical vertebrae, 4 cases were in thoracic vertebrae,and 44 cases were in lumbar vertebrae. Thirty patients with spinal fracture were caused by trauma and 33 patients without trauma, 11 patients combined with nerve injury. The patients were divided into two groups according to plasma D-dimer levels, more than or equal to 500 microg/L was D-dimer positive group and less than 500 microg/L was D-dimer negative group. Venous blood of all patients early morning with empty stomach were testd on admission, and at 2 h, 1 d, 2 d, 3 d, 4 d, 6 d, 8 d, 10 d, 15 d after operation,respectively.

RESULTSThere was no statistically significant differences in sex, operative segments, implants, operative posture, age, bleed volume, body weight, peroperative D-dimer levels between two groups. After operation, plasma D-dimer of 19 patients were more than or equal to 500 microg/L, with persistent or progressive increasing. Two cases occurred deep venous thrombosis in D-dimer positive group, they respectively were found at 3 days and 8 days after operation. Both of them underwent posterior decompression and internal fixation. However,no deep venous thrombosis was found in D-dimer negative group.

CONCLUSIONPostoperative D-dimer assay can effective predict deep venous thrombosis occurrence. D-dimer level more than or equal to 500 microg/L will be considered as a risk factor for deep venous thrombosis after spinal surgery.

Adult ; Aged ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spine ; surgery ; Ultrasonography ; Venous Thrombosis ; blood ; diagnostic imaging ; surgery ; Young Adult

OBJECTIVETo study the protective effect of dl-tetrahydropalmatine(dl-THP) on liver injury induced by carbon tetrachloride (CC4) in mice.

METHODMice were administracted with dl-tetrahydropalmatine ip 20, 40 mg x kg(-1) daily for 9 d respectively, and then actue liver injury model was induced by 0.1% carbon tetrachloride ip 20 mL x kg(-1). The mice were killed 17 h after injection ip of CCl4, serum alanine and aspartate aminotransferase (ALT and AST) activity were measured, and maleic dialdehyde (MDA) and superoxide dismutase(SOD) activity in liver were detected.

RESULTdl-THP significantly reduced the level of serum ALT and AST, inhibited lipoperxidation in liver, while increased SOD activity in liver tissue. Degeneration of hepatocytes was obviously prevented in mice treated with dl-THP, and the liver histological structure was well maintained.

CONCLUSIONdl-THP has inhibitory effects on liver injury induced by CCl4 in mice. The mechanisms may be related with its effects of reducing lipid peroxidation product.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Berberine Alkaloids ; pharmacology ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; etiology ; metabolism ; pathology ; Female ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Protective Agents ; pharmacology ; Random Allocation ; Superoxide Dismutase ; metabolism

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Drug Synergism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Transfection

OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

This study was aimed to explore the inhibitory effect of triptolide on proliferation and inducing apoptosis effect of K562/G01 cells and their possible mechanism. MTT assay was used to detect the effect of imatinib or triptolide alone and their combination on K562/G01 proliferation; the cell cycle, apoptosis rate, P-gp protein expression were detected by flow cytometry (FCM); the expression of P-gp was assessed by Western blot; the BCR/ABL gene expression was assayed by real time quantitative PCR. The results showed that triptolide could enhance the effect of imatinib on proliferation inhibition and apoptosis of K562/G01, arrested the cell cycle in G1 phase, down-regulated the expression of BCR/ABL gene and P-gp protein. It is concluded that triptolide induces K562/G01 cell proliferation inhibition and apoptosis, the mechanism may be related to cell cycle arrest, decrease of P-gp protein expression, inhibition of BCR/ABL gene expression.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Apoptosis ; drug effects ; Benzamides ; pharmacology ; Cell Cycle Checkpoints ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Drug Resistance, Neoplasm ; Epoxy Compounds ; pharmacology ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Imatinib Mesylate ; K562 Cells ; Phenanthrenes ; pharmacology ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

Background and purpose:Adenosine triphosphate-binding cassette superfamily G member 2 (ABCG2), which has been found over-expressed in a variety of cancer cells, takes part in the drug resistance of cancer through effux of anticancer drugs. The purpose of this study was to investigate the mechanisms of human glioblastoma cells sensitivity to pyropheophorbide-a methyl ester (MPPa)-mediated photodynamic therapy (PDT) eradicating tumour cells and its relationship to ABCG2.Methods:U87 and A172 glioma cell lines in the logarithmic growth phase were selected and exposed to the treatment of MPPa-PDT and MPPa-PDT+fumitremorgin C (FTC) respectively. The cell viability was measured with the use of CCK-8 assay. The expression of ABCG2 was detected by Western blot. The intracellular contents of MPPa in each group without illumination were tested by lfow cytometry. Flow cytometry with AnnexinⅤ-FITC/PI double staining was used to detect the cell apoptotic rate. DCFH-DA staining was used to assess the generation of intracellular reactive oxygen species (ROS).Results:The MPPa-mediated PDT could eradicate A172 and U87 cancer cells in an energy-dependent manner. The light energy density in A172 was 8 times of that in U87 when the cell viability reached median lethal dose after MPPa-mediated PDT. The high expression of ABCG2 in A172 cells affected the accumulation of intracellular MPPa. Inhibition of ABCG2, not only could enhance the eradicating effect of MPPa-PDT on A172 cells, but also could increase the yield of ROS triggered by MPPa-PDT and the accumulation of intracellular MPPa.Conclusion:The human glioblastoma cell line A172 is insensitive to MPPa-mediated PDT. The mechanism may relate to ABCG2, which decreases the MPPa content in cancer cells through effux of MPPa, resulting in decline of cytotoxicity.

Objective To analysis the E protein epitopes of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus and to distinguish the shared or specific epitopes among them. Methods Bioinformatic software DNAStar was used to analyze the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E prtein amino acid sequences. The influence of secondary structure was also considered. Based on the bio-informatic analysis of E protein epitopes, 6 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-c2x. The vectors was then transferred into E.coli BL21 (DE3) and Rosetta (DE3). Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of gene segments and SDS-PAGE were used identify the expression proteins. The antigenieity was tested, using Western blot. Results 15 shared epitopes and 47 specific epitopes were forecasted by bioinformatic analysis, and 6 specific epitopes from dengue virus type 1-4, Japanese encephalitis virus and yellow fever virus E protein were expressed in E.coli successfully. Two specific antigenic determinant from dengue virus type 1 and dengue virus type 2 were confirmed using Western blot, while the others epitopes shown no antigenic reaction property. Conclusion Two specific antigenic determinant were confirmed, under Western blot.

OBJECTIVETo prepare rat whole-kidney acellular matrix (ACM) scaffolds using fluid perfusion method.

METHODSThe kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH2O. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining.

RESULTSNo cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting of basilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope.

CONCLUSIONFluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.

Animals ; Biocompatible Materials ; Cell Separation ; methods ; Extracellular Matrix ; Female ; Kidney ; cytology ; Male ; Perfusion ; Rats ; Rats, Wistar ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo explore the effects of pre-acupuncture and immediate acupuncture on kidney function and oxygen free radical metabolism in rats with simulated weightlessness.

METHODSTwenty male clean-grade Wister rats were randomly divided into a normal control group, a model group, a pre-acupuncture group and an immediate acupuncture group, 5 rats in each one. The rats in the normal control group did not receive any treatment but free activities for 4 weeks. The rats in the rest groups received 4-week tail suspension to establish the model of simulated weightlessness. One week before the tail suspension, the rats in the pre-acupuncture group were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once a day for 7 days. The rats in the immediate acupuncture group received tail suspension and acupuncture at the same time; during the tail suspension, the electroacupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and "Sanyinjiao" (SP 6) for 30 min per treatment, once every other day for 14 days. The colorimetric method was used to measure the content of blood urea nitrogen (BUN) in serum as well as activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) and content of malonaldehyde (MDA) in renal tissue in each group.

RESULTSCompared with the normal control group, the content of BUN in the model group was increased significantly (P<0.01), the activity of SOD and GSH-PX in nephridial tissue was significantly reduced (both P<0.01), and the content of MDA was increased significantly (P<0.05). Compared with the model group, the content of BUN in the pre-acupuncture group and immediate acupuncture group was significantly reduced (P<0.01, P<0.05), the activity of GSH-PX in the pre-acupuncture group was obviously increased (P<0.05) and the content of MDA in the immediate acupuncture group was increased significantly (P<0.01). Compared with the immediate acupuncture group, the content of MDA in the pre-acupuncture group was lower (P<0.01).

CONCLUSIONThe pre-acupuncture and immediate acupuncture both have the capacity to improve the kidney function and anti-oxygen free radical injury in rats with simulated weightlessness, however, the capacity to increase the protection ability of the kidney and eliminate free radical in the pre-acupuncture group is superior to that in the immediate acupuncture group, which is likely to be related with improving antioxidant ability of kidney.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Antioxidants ; metabolism ; Humans ; Kidney ; metabolism ; Kidney Diseases ; metabolism ; physiopathology ; therapy ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Space Flight ; Superoxide Dismutase ; metabolism ; Weightlessness ; adverse effects

Objective Circulating tumor cells (CTCs) have potential value in the clinical application of various tumors. This study was to investigate the role of CTCs and their chemokine receptor CCR9 in the invasion and metastasis of non-small cell lung cancer (NSCLC). Methods From May 2018 to June 2019, a total of 62 patients with NSCLC in the clinical oncology center of The People's Hospital of Guangxi Zhuang Autonomous Region were enrolled in this study. The CanpatrolTM CTC technique was used to detected the expressions of CTCs and CCR9 in CTCs in peripheral blood of patients. Furthermore, the relationships between expression levels of CTCs, CCR9 and clinical, pathological characteristics of NSCLC patients were analyzed. Results CTCs were detected in 56 of 62 (90.3%) NSCLC patients. CTCs counts were associated with TNM stage, lymph node metastasis and distant metastasis of NSCLC (P<0.05). In the analysis of clinical correlation between CTC subtypes and NSCLC, epithelial CTCs counts were related to TNM stage and distant metastasis of NSCLC (r=0.296 and r=0.273, P<0.05). Additionally, counts of mixed type CTCs were also correlative with NSCLC tumor metastasis (r =0.253, P =0.047). Finally, we found that the positive rate of CCR9 in mixed type CTCs was associated with distant metastasis of NSCLC (r=0.353, P=0.038). Conclusion CTCs counts and subtypes were correlated with TNM stage and metastasis of NSCLC. The expression level of CCR9 on CTC was expected to be a biomarker to evaluate the risk of tumor metastasis in NSCLC.