BACKGROUND:The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are increased during infectious brain edema, and are positively relevant to the degree of brain damage. However, whether TNF-α can enhance blood brain barrier (BBB) permeability remains unclear, especially in vitro.OBJECTIVE: To understand the changes and possible mechanism of the BBB permeability induced by TNF-α in vitro.DESIGN: Randomized controlled cell model study in vitro.SETTING:Department of Pediatrics, Xiangya Hospital, Central South University; Department of Biochemistry, Xiangya Medical College, Central South University.MATERIALS: Twenty 7-day-old healthy Sprague-Dawley rats, of clean grade and either gender, were provided by the Animal Center, Xiangya Hospital, Central South University. TNF-α was purchased from sigma Company; DMEM fluid medium and fetal bovine serum were purchased from Hyclone Company; Y-27632 was purchased from Alexis Company,and rabbit anti-human factor Ⅷ -related antigen was purchased from Zymed Company; Mouse anti-rat glial fibrillary acidic protein (GFAP) was purchased from Neomarkers. Other biochemical reagents were imported (Sigma Company).METHODS: This experiment was carried out in the Xiangya Hospital, Central South University between March 2004 and April 2005. Brain microvascular endothelial cells and astrocytes were co-cultured 10 days to set up rat models of BBB in vitro. Then, the cells were divided into 4 groups: model group(BBB models were prepared), TNF-α group ( BBB model incubated with 0.01 g/L TNF-α for 5 hours), Y-27632 pretreated group ( BBB model incubated with 30 μmol/L Y-27632 for 1 hour before 0.01g/L TNF-α challenge ) and Y-27632 control group (BBB models only incubated with Y-27632 as those in the Y-27632 pretreated group). The effect of TNF-α on BBB permeability was observed by detecting the 125 I -BSA, which passed through the inserts at each time point (30,60,120 and 240 minutes) using .γradioimmunoassay counter.MAIN OUTCOME MEASURES: The 125 I -BSA, which passed through the inserts in rat models of BBB at different time points after intervention.RESULTS: The 125 I -BSA, which passed through the inserts in rat models of BBB, was all significantly higher in the TNF-α group than in the other groups at 30, 60, 120 and 240 minutes after intervention, respectively (P ＜ 0.01), and reached the peak at 240 minutes; The 125 I -BSA, which passed through the inserts, was lower in the Y-27632 pre-treated group than in the TNF-α group at 30 and 60 minutes after intervention (P＜ 0.01). There was also significant difference in 125 I -BSA permeation between Y-27632 pretreated group and Y-27632 control group after 120 minutes (P ＜ 0.05).CONCLUSION: TNF-α can increase BBB permeability, and Y-27632 pretreatment can early reverse the effect of TNF-α on BBB permeability.

Hydrophidae, one of the precious traditional Chinese medicines, is generally drily preserved to prevent corruption, but it is hard to identify the species of Hydrophidae through the appearance because of the change due to the drying process. The identification through analysis on gene barcode, a new technique in species identification, can avoid the problem. The gene barcodes of the 6 species of Hydrophidae like Lapemis hardwickii were aquired through DNA extraction and gene sequencing. These barcodes were then in sequence alignment and test the identification efficency by BLAST. Our results revealed that the barcode sequences performed high identification efficiency, and had obvious difference between intra- and inter-species. These all indicated that Cyt b DNA barcoding can confirm the Hydrophidae identification.
Animals ; Base Sequence ; China ; Cytochromes b ; genetics ; DNA Barcoding, Taxonomic ; Elapidae ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

Objective To understand the changes and possible mechanism of the blood brain barrier(BBB) permeability induced by tumor necrosis factor(TNF-?) in vitro.Methods BBB model was established by coculturing allogenic brain microvessel end othelial cell(BMEC) and astrocyte(AS).The BBB model was divided randomly into normal control group,TNF-? group and Y-27632 pretreatment group.The changes of BBB permeability were evaluated by Gamma radioim muno assay counter.Results The Gamma radioimmuno assay indicated that the marker,~(125)I-BSA,across the BBB model in vitro was significantly increased after TNF-? treatment compared with control group,Y-27632 pretreatmen could prevent the permeability of BBB induced by TNF-?(P

Objective:To explore the influence of ulinastatin combined with thymosinα1 on the immune function of patients with a-cute brain injury. Methods:Sixty-eight cases of patients with acute brain injury were divided into the control group and the observation group randomly with thirty-four ones in each. The control group was given the routine treatment, and the observation group was given ulinastatin combined with thymosinα1 additionally. After the 1-day, 3-day, 7-day and 14-day treatment, transforming growth factor-β1 (TGF-β1), interleukin-6(IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) and the other serum inflammatory cyto-kine levels, and CD4 +, CD8 +, CD4 + /CD8 +, HLA-DR and cellular immune index levels were detected in the two groups. The prog-nosis effects were evaluated by the prognostic classification of brain injury, and the adverse reactions were analyzed in the two groups as well. Results:After the 1-day treatment, there were no significant differences in the serum inflammatory cytokines and immune param-eters between the groups (P>0. 05). After the 3-day, 7-day and 14-day treatment, serum TGF-β1, IL-6, IL-10 and TNF- α levels were higher than those on the first day after the treatment, and TGF-β1 showed an increasing trend with time extension, while IL-6, IL-10, TNF-α and CD4 + and CD4 + /CD8 +rose first and then decreased. After the 3-day, 7-day and 14-day treatment, serum IL-10 and CD4 +levels in the observation group were significantly higher than those in the control group, and IL-6 and TNF-αlevels were signifi-cantly lower than those in the control group (P<0. 05). After the 3-day and 14-day treatment, CD4 + and CD8 + levels in the observa-tion were significantly higher than those in the control group, and after the 7-day and 14-day treatment, HLA-DR levels were signifi-cantly higher than those in the control group (P<0. 05). The prognosis effect of the observation group was better than that of the con-trol group with statistically significant difference (P<0. 05). Conclusion:Ulinastatin combined with thymosin α1 is used to treat the patients with acute brain injury with better cellular immune function improvement and prognosis effect, which is worthy of clinical popu-larization and application.

Modern pharmacological studies have shown that Cistanche deserticola (C. deserticola) has a protective effect on the liver, but its active fraction and mechanism are not clear. In order to identify the effective fraction of C. deserticola Y. C. Ma, an acute alcoholic liver injury model in mice was established with 56-proof Erguotou and different fractional extracts of C. deserticola Y. C. Ma (total glycosides, polysaccharides, and oligosaccharides) were administered. After 14 days of oral administration, liver pathology and lipid deposition were measured and the expression of nuclear factor E2-related factor (Nrf-2), kelch-like ECH-associated protein-1 (Keap-1), and plasmalemma vesicle-associated protein-1 (PV1) were measured by immunofluorescence. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), endotoxin (ET), diamine oxidase (DAO), and D-lactic acid (D-LA) in serum, and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and malondialdehyde (MDA) in liver were measured by ELISA. All animal experiments were carried out with approval of the Experimental Animal Welfare Ethics Committee of the Peking University Health Science Center. The results show that the total glycosides of C. deserticola Y. C. Ma (400 mg·kg^-1) could decrease liver pathology, decrease serum endotoxin, diamine oxidase, and D-lactic acid, and reduce hepatic lipid deposition. Total glycosides also promoted Nrf-2 transfer into the nucleus and decreased the expression of Keap-1 and PV1. In summary, the total glycosides of C. deserticola Y. C. Ma had a protective effect in acute alcoholic liver injury and the mechanism may be related to the activation of the Nrf-2/Keap-1 pathway, improvement of intestinal wall integrity, and inhibition of the transport of harmful substances into the liver.

ObjectiveTo investigate the relationship between the serum concentration of IL-6,S100β,NT-3 and the cognitive functions in first-episode schizophrenia characterized by positive or negative symptoms.Methods44 first-episode schizophrenic patients characterized by positive symptoms (positive group),36 first-episode schizophrenic patients characterized by negative symptoms (negative group) and 50 healthy controls (controls) were collected.The serum levels of IL-6,S100β and NT-3 were measured by enzyme-linked immunosorbent assay (ELISA).The systematic evaluation tool-MCCB was applied to assess cognitive function in patients and controls.ResultsNT-3 serum levels in positive or negative groups were lower than those in controls and the differences were significant((118.39±37.50) ng/L,(112.55±32.29) ng/L vs (141.18±29.67) ng/L) (P<0.01).IL-6 and S100β serum levels in positive or negative groups were higher than those in controls and the differences were statistically significant((5.74±1.00)ng/L,(5.07±1.17)ng/L vs (4.23±0.91)ng/L),((132.98±46.71)ng/L,(124.99±43.14)ng/L vs (103.63±31.57)ng/L)(P<0.01).IL-6 serum levels in the positive group ((5.07±1.17)ng/L) were lower than those in the negative group ((5.74±0.99)ng/L) and the difference was statistically significant (P<0.05).In MCCB test,the TMT scores in patients characterize by positive symptoms or patients characterize by negative symptoms were higher than those in healthy control group (P<0.01).BACS SC,HVLT-R WMS-Ⅲ,SS,NAB,BVMT-R,CF in patients characterize by positive symptoms or by negative symptoms were lower than those in healthy control group(P<0.01).There were no statistical difference in the MCCB scores between the patients with positive symptoms and negative symptoms.In positive group,there was a positive correlation between the IL-6 serum concentration and the general symptom scores in PANSS (P<0.05).In positive group,NT-3 serum concentration was positively correlated with the general symptom scores or total scores of PANSS (P<0.05).BVMT-R scores in MCCB were also positively correlated with IL-6 or NT-3 serum concentration in positive group (P<0.05).ConclusionThe impairment of part of cognitive functions for schizophrenic patients may be related to the serum protein factors.There may be different in pathophysiology between the first-episode schizophrenic patients characterized by positive symptoms and those characterized by negative symptoms.

Objective To investigate the influence of aspirin and/or clopidogrel treatment on platelet aggregation rate in coronary heart disease (CHD) patients,and discuss the factors related to anti-platelet drug resistance.Methods A total of 160 patients with CHD and received aspirin and/or clopidogrel treatment were enrolled in the Second Xiangya Hospital,Central South University,and were divided into stable coronary heart disease (SCHD) group (n =90) and acute coronary syndrome (ACS) group (n =70).Meanwhile,non-coronary heart disease (NCHD) patients who did not receive anti-platelet drug treatment were enrolled as controls (n =50).Clinical data of the subjects were recorded.The maximum platelet aggregation rate induced by arachidonic acid (MAR-AA) and adenosine diphosphate (MAR-ADP) were evaluated with sequential platelet counting method.The factors related to drug resistance were analyzed with Logistic regression analysis.Results Compared to NCHD group,there were lower MAR-AA and MAR-ADP in two groups of CHD (all P ＜ 0.05).In ACS patients,MAR-AA and MAR-ADP are significantly lower (P ＜0.05) in patients who receive the aspirin and clopidogrel.The rate of anti-platelet drug resistance in ACS group was significantly higher than that in SCHD group (20.0％ vs 10.0％,P ＜ 0.05).Multivariate logistic regression analysis showed that low HDL-C (＜ 1.0 mmol/L) was an independent risk factor related to the anti-platelet drug resistance (OR =4.36,95 ％ CI:1.36-14.02,P =0.025).Conclusions The antiplatelet treatment with aspirin and/or clopidogrel can significantly reduce the platelet reactivity in CHD patients,but some patients still present anti-platelet drug resistance.The combination of aspirin and clopidogrel is better.The rate of drug resistance in ACS patients is high.Low HDL-C might be associated with anti-platelet drug resistance.

OBJECTIVETo investigate the features and causes of complications of unilateral pedicle screw fixation combined with contralateral percutaneous translaminar facet screw fixation and lumbar interbody fusion in treating lower lumbar diseases.

METHODSThe clinical data of 166 patients with lower lumbar diseases who underwent unilateral pedicle screw fixation combined with contralateral percutaneous translaminar facet screw fixation and lumbar interbody fusion with intervertebral cages from January 2008 to December 2013 were retrospectively analyzed. There were 64 males and 102 females, aged from 24 to 74 years with a mean of 51.9 years old, suffered from lower lumbar lesions for 47.5 months on average (ranged, 8 months to 30 years). Among these patients, lumbar intervertebral disc degeneration was found in 49 patients, recurred lumbar intervertebral disc protrusion in 17 patients, massive lumbar intervertebral disc protrusion in 23 patients, lumbar intervertebral disc protrusion accompany with spinal canal stenosis in 27 patients, lumbar degenerative spondylolisthesis with degree I (Meyerding grade) in 21 patients, far lateral lumbar intervertebral disc protrusion in 5 patients. Single segmental diseases occurred in 124 patients and two segmental diseases in 42 patients. The diseases occurred at L(3,4) segment in 6 patients, at L(4,5) segment in 97 patients, at L5S1 segment in 21 patients, at L(2,3), and L(3,4) segments in 1 patient, at L(3,4) and L4,5) segments in 26 patients, and at L(4,5), and L5S1 segments in 15 patients.

RESULTSThere was no abnormal bleeding in the patients and no patient received blood transfusion. During the surgery, spinal dura mater injury with cerebrospinal fluid leakage complicated in 1 patient, a fracture of vertebral pedicle in 4 patients, and end plate injury in 2 patients. No postoperative cerebrospinal fluid, incision infection and skin necrosis were found after operation. Nerve root injury was found in 1 patient. According to the position of pedicles crew, 371 screws of 163 patients were in degree I and 3 screws of 3 patients were in degree II; position of translaminar facet screw, 199 screws of 157 patients were type I, 8 screws of 8 patients were type II, 1 screw of 1 patient was III. Translaminar facet screw was slightly short in 2 patients. Five patients were lost to follow-up, two patients were died. The remaining patients were followed up for 35.4 months on average (ranged, 12 to 60 months). During the follow-up period , end plate was cut off and intervertebral cages were embedded in 14 segments of 14 patients. Abnormal pain of both lower extremities was found in 1 patient. With the exception of 11 unidentified segments in 11 patients, 189 segments of 148 patients obtained intervertebral fusion. No loosening, displacement, breakage of pedicle screw or translaminar facet screw, displacement of intervertebral cages or obvious degeneration of adjacent segments were found. The coronal and sagittal planes balance of lumbar vertebra were obviously improved. Postoperative JOA score was significantly increased than that of preoperative.

CONCLUSIONUnilateral pedicle screw fixation combined with contralateral percutaneous translaminar facet screw fixation and lumbar interbody fusion with intervertebral cages is a good choice for the treatment of lower lumbar diseases, but it has a risk of complications. Abundant surgeon's surgical experience, careful operation, and rational use of imaging technique can effectively reduce the incidence of complications.

Adult ; Aged ; Bone Plates ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Degeneration ; surgery ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Retrospective Studies ; Spinal Fusion ; Spondylolisthesis ; surgery ; Treatment Outcome ; Young Adult

Laser scanning confocal microscopy (LSCM) and whole-cell perforated patch-clamp techniques were combined to study simultaneously the changes of intracellular signal molecules and membrane currents. Intracellular calcium transients and spontaneous transient outward currents (STOCs) were recorded simultaneously in freshly isolated mouse cerebral artery smooth muscle cells. The cells loaded with fluo-4/AM were scanned with the confocal line-scan mode. Triggering voltage pulses derived from an EPC-10 patch clamp amplifier triggered the confocal line scan. The results showed that STOCs and intracellular calcium transients could be simultaneously recorded in the same cell. This technique will be useful in studies of diseases caused by impairments of intracellular Ca(2+) signaling and related ionic channel activities, or vice versa.
Animals ; Calcium Signaling ; Cerebral Arteries ; cytology ; Mice ; Myocytes, Smooth Muscle ; physiology ; Patch-Clamp Techniques

OBJECTIVETo investigate the safety and feasibility of glans-preserving surgery in the treatment of superficial penile cancer (SPCa).

METHODSWe retrospectively analyzed the clinical data of 21 cases of SPCa treated by glans-preserving surgery in our hospital from January of 2003 to March of 2010.

RESULTSThe study included 21 SPCa patients aged 36 to 57 (mean 46) years, with superficial lesions involving the glans penis, coronary sulcus or shaft skin. The tumors were staged and graded TaG1 in 6 cases, TaG2 in 5, TisG1 in 2, TisG2 in 4, T1G1 in 3, and T1G2 in 1. All the patients underwent glans-preserving surgery to preserve the normal appearance and functional integrity of the glans penis, and all returned to normal sexual activity 1 month after operation, with good sexual function and sexual satisfaction. Postoperative follow-up lasted 2 to 7 (mean 5) years, and 2 cases of recurrence in situ found at 6 and 9 months, respectively, which were successfully managed by a second glans-preserving surgery.

CONCLUSIONGlans-preserving surgery is an effective method for superficial penile cancer. With proper selection of the patients, this procedure is technically safe, maximally preserves the penile appearance, and least affects the patients' sexual satisfaction.

Adult ; Carcinoma, Squamous Cell ; surgery ; Humans ; Male ; Middle Aged ; Penile Neoplasms ; surgery ; Penis ; surgery ; Retrospective Studies ; Treatment Outcome