[Clinical study of application minimally invasive expandable channel in lumbar discectomy and interbody fusion and internal fixation].

OBJECTIVE: To explore the advantages of minimally invasive expandable in surgery of lumbar discectomy and interbody fusion and internal fixation. METHODS: The clinical data of 48 patients who underwent lumbar discectomy and interbody fusion and internal fixation from January 2010 to March 2016 was retrospectively analyzed. According to the admission queue, the patients were randomly assigned into channel group (26 cases) or traditional group (22 cases). In channel group, surgical approach of minimally invasive expandable channel was applied, and in traditional group, open posterior operation approach (including posterior lumbar interbody fusion and transforaminal lumbar interbody fusion, etc.) was applied. In channel group, there were 20 males and 6 females, aged from 43 to 74 years with an average of(56.6±5.1) years; course of disease was ranged from 4 to 22 months with an average of (6.7±1.8) months; 1 case was complicated with diabetes, 6 cases were complicated with hypertensive disease, and 2 cases were complicated with arrhythmia. In traditional group, there were 15 males and 7 females, aged from 43 to 73 years with an average of(55.9±4.6) years; course of disease was ranged from 4 to 26 months with an average of (6.2±2.1) months; 2 cases were complicated with diabetes, 5 cases were complicated with hypertensive disease, and 1 case was complicated with arrhythmia. Operation time, bleeding volume, and hospitalization time were compared between two groups and visual analogue scale(VAS), Oswestry Disability Index(ODI), bone fusion information, and complications correlated with incision were observed in two groups. RESULTS: All 48 patients were followed up for more than 6 months. Postoperative VAS and ODI were significantly improved (P<0.01), but 3 and 6 months after operation, there was no significant difference in VAS between two groups, and ODI score of channel group was lower than that of traditional group(P<0.01). Operation time, bleeding volume, hospitalization time in channel group respectively were (167.3±30.2) min, (786.8±147.8) ml, (12.3±2.4) d, and in traditional group were (197.5±48.7) min, (786.8±147.8) ml, (16.5±3.8) d, there was significant differences between two groups. There was no significant difference in fusion rate and fusion time between two groups. There were 4 cases and 7 cases developed incision related complications in channel group and traditional group, respectively. The difference between two groups was significant(P<0.01). CONCLUSIONS: Compared with conventional surgery minimally invasive lumbar discectomy and interbody fusion and internal fixation has advantages of less trauma, shorter operative time and better functional recovery.

Potential Role of Hepatitis C Virus Alternate Reading Frame Protein in Negative Regulation of T-Bet Gene Expression.

Hepatitis C virus (HCV) is a major cause of chronic liver disease and has led to cirrhosis or hepatocellular carcinoma in a majority of infected individuals. We have previously demonstrated that the HCV alternate reading frame protein (F protein) is related to Th1/Th2 bias in chronic hepatitis C (CHC) patients, and we aimed to explore the relative molecular mechanisms here. A total of 104 cases including CHC patients and healthy donors were enrolled. T-bet and GATA-3 expression levels were analyzed in peripheral blood mononuclear cells (PBMCs). The levels of signal transducer and activator of transcription-1/-6(STAT1/6) and phosphorylated STAT1/6(pSTAT1/6) in PBMCs were measured by Western blotting. Our results showed that the levels of T-bet in PBMCs, as well as the levels of gamma interferon (IFN-γ) in sera, were decreased in anti-F protein antibody seropositive patients compared with anti-F protein antibody seronegative patients, whereas the levels of GATA-3 did not show difference between the two groups. Moreover, the decreased pSTAT1 and increased pSTAT6 were observed in PBMCs by HCV core/F protein stimulation with constant STAT1/6 expression. Taken together, it suggested that T-bet may be involved in Th1/Th2 bias induced by HCV F protein, and the disruption of STAT phosphorylation may participate in this mediation.

Inhibition of ATP-induced glutamate release by MRS2179 in cultured dorsal spinal cord astrocytes.

It was reported that ATP, an excitatory chemical mediator, exerts its effects by activation of the P2X (ligand-gated cationic channels) and P2Y (G protein-coupled receptors) purinoceptors in the nervous system. In the present work, we used confocal laser scanning microscopy and high-performance liquid chromatography to assess the role of the P2Y1 receptor in ATP-evoked Ca2+ mobilization and glutamate release from cultured dorsal spinal cord astrocytes. ATP (0.01-100 micromol/l) produces a dose-dependent rise in the Ca2+ relative fluorescence intensity in cultured astrocytes. N6-methyl-2'-deoxyadenosine-3',5'-bisphosphate (MRS2179, 0.01-100 micromol/l), a P2Y1-specific antagonist, could dose-dependently inhibit ATP-evoked Ca2+ mobilization. In addition, 100 micromol/l ATP caused glutamate efflux from cultured dorsal spinal cord astrocytes in a time-dependent manner. 100 micromol/l MRS2179 significantly inhibited the glutamate efflux induced by ATP, which suggests that P2Y1 receptor activation is responsible for the ATP-induced glutamate efflux from astrocytes. Taken together, our results demonstrate that P2Y1 receptor plays an important role in modulating the function of astrocytes, which raises the possibility that MRS2179, a potent P2Y1-specific antagonist, may become a potential drug in treating many chronic neurological diseases characterized by astrocytic activation in the nervous system.

P2Y1 receptor-mediated glutamate release from cultured dorsal spinal cord astrocytes.

P2 receptors have been implicated in the release of neurotransmitter and proinflammatory cytokines by the response to neuroexcitatory substances in astrocytes. In the present study, we examined the mechanisms of ADP and adenosine 5'-O-2-thiodiphosphate (ADPbetaS, ADP analogue) on glutamate release from cultured dorsal spinal cord astrocytes by using confocal laser scanning microscopy and HPLC. Immunofluorescence activity showed that P2Y(1) receptor protein is expressed in cultured astrocytes. ADP and ADPbetaS-induced [Ca(2+)](i) increase and glutamate release are mediated by P2Y(1) receptor. Ca(2+) release from IP(3)-sensitive calcium stores and protein kinase C (PKC) activation is important for glutamate release from astrocytes. Furthermore, P2Y(1) receptor-evoked glutamate release is regulated by volume-sensitive Cl(-) channels and anion co-transporter, which open up the possibility that P2Y(1) receptor activation causes the increase of cell volume. Release of glutamate by ADPbetaS was abolished by 5-nitro-2 (3-phenyl propy lamino)-benzoate plus furosemide but was unaffected by botulinum toxin A. These observations indicate that P2Y(1) receptor-evoked glutamate may be mediated via volume-sensitive Cl(-) channel but not via exocytosis of glutamate containing vesicles. We speculate that P2Y(1) receptors-evoked glutamate efflux, occurring under pathological condition, may modulate the activity of synapses in spinal cord.

[Progress of studies on multidrug resistance in hematological malignancies--review].

Chemotherapy remains a major route of intervention in hematological malignancies. However, a key issue in the treatment of hematological malignancies is the development of multidrug resistance (MDR) to chemotherapeutic drugs. Several mechanisms may account for this phenomenon, including biochemical mechanisms, such as the overexpression of resistance-conferring proteins and physiological mechanisms involving the hematopoietic microenvironment. In this article the pathomechanism, diagnostic approach, interpretation of results from clinical samples and correlations with hemopoietic microenvironment were briefly reviewed. The aspects of development and problems in MDR study as well as detection methods for MDR were also discussed.

[Construction and expression of adeno-associated virus vectors of Smad 6 and Smad 7 genes in human renal tubule epithelial cells].

AIM: To construct the adeno-associated virus(AAV) vectors of Smad 6 and Smad 7 genes and observe their expressions in human renal tubule epithelial cells. METHODS: The plasmids pcDNA3-Smad 6/flag and pcDNA3-Smad 7/flag were digested with BamH I and Xho I. Then the Smad 6/flag and Smad 7/flag gene fragments were cloned into plasmid pAAV-MCS, respectively to construct the recombinant pAAV-Smad 6/flag and pAAV-Smad 7/flag plasmids. The recombinant expression plasmid or pAAV-LacZ plasmid were co-transfected into the HEK 293 cells with pHelper and pAAV-RC by calcium-phosphate precipitation method. Recombinant AAV-2 viral particles were prepared from infected HEK293 cells and then were used to infect human renal tubule epithelial cells (HKCs), The expressions of Smad 6 and Smad 7 in HKCs were demonstrated by immunocytochemical staining. RESULTS: The recombinant AAV vectors of Smad 6 or Smad 7 genes were constructed and expressed in the HKCs successfully. CONCLUSION: These results indicate that AAV can deliver Smad 6 and Smad 7 genes to renal cells in-vitro, suggesting the recombinant AAV can be used for gene therapy of renal fibrosis.

Expression of an antitumor-analgesic peptide from the venom of Chinese scorpion Buthus martensii karsch in Escherichia coli.

The gene encoding a putative mature antitumor-analgesic peptide (AGAP) from the venom of the Chinese scorpion Buthus martensii Karsch was obtained by polymerase chain reaction (PCR) according to its cDNA sequence and expressed in Escherichia coli. While most of the recombinant AGAP was expressed in the form of insoluble inclusion body. The recombinant AGAP was purified to homogeneity by metal chelating affinity chromatography. Pharmaceutical tests showed that the recombinant AGAP has both analgesic and antitumor activities on mice.