Objective To investigate the clinical value of stereotactic biopsy in the diagnosis of the multiple intracranial lesions, and explore the operation methods, technical points and clinical experiences to reduce surgical complications. Methods Twenty-three patients in the first affiliated hospital of Kunming Medical University underwent stereotactic biopsy from January 2009 to June 2013 were analyzed retrospectively. The patients were aged between 11 and 73 years (the mean age of 34.6 years) . There were 12 males and 11 females. Operations were performed by thin thickness of spiral CT scan, ASA-602S and Leksell-Frame-G stereotactic frame, Sedan side-cutting needle, Backlund side-cutting needle and neuroendoscope of STORZ. Routine histopathological examinations of specimens were conducted. The immunohistochemical staining of the histopathological section of specimens was performed if necessary. Results The diagnostic yield was 91.3% (21 cases) . The result of pathological diagnosis was inflammatory granuloma in 1, inflammatory lesion in 1, calcification in 1, hyperplasia of colloid cells in 6, astrocytoma in 1 (WHOⅠ-Ⅱ), astrocytoma in 2 (WHOⅡ), astrocytoma in 1 (WHOⅡ-Ⅲ), mixed oligoastrocytoma in 1 (WHOⅠ-Ⅱ), glioblastoma multiforme in 3 (WHOⅣ), germinoma in 1, brain metastases in 1, diffuse large B-cell lymphoma in 1, intracranial granulomatousarteritis in 1 and negative in 2. There were no serious complications, such as coma, hemiparalysis, infection and intracranial hematoma. Conclusion Stereotactic biopsy is an important method in the diagnose of multiple intracranial lesions. It has the advantages of precise location, less damage, safe performance, and reducing the complication effectively. It is worth promoting.

Objective To investigate the influence of MgSO4 with different concentrations on cell survival and γ-H2AX expression in HUVEC irradiated with X-rays.Methods Cell proliferation rate was assayed by CCK-8,γ-H2AX foci formationwas observed with a laser confocal microscope,and γ-H2AX protein expression was detected by flow cytometer and Western blot assay.Results MgSO4 with a concentration of 1.25 mg/ml could improve the survival rate of IHUVEC treated with X-rays (t =-6.34,P ＜ 0.05).After irradiation,γ-H2AX loci approached to the highest level from 30 min to 1 h after radiation and then decreased.MgSO4 significantly reduced the foci formation at 0.5,1,2,6 and 12 h postirradiation (t =12.62,6.36,11.93,5.75,9.43,P ＜ 0.05).Both flow cytometry,and Western blot assays showed that MgSO4 inhibited γ-H2AX protein expression at 0.5,1 and 2 h post-irradiation (t =6.07,5.32,11.85,P ＜ 0.05).Conclusions MgSO4 could improve the survival rate and reduced γ-H2AX expression in HUVEC irradiated with X-ray.

ObjectiveTo investigate the expression of miR-155 and miR-146a in peripheral blood mononuclear cells (PBMC) and plasma of rheumatoid arthritis (RA) patients.MethodsPBMC and plasma were separated from the peripheral blood of 34 RA patients and 15 healthy individuals.Total RNAs were isolated and miRNAs were purified.The levels of miR-155 and miR-146a were determined by quantitative reverse transcription PCR(qRT-PCR).U6 was used as housekeeping control.The amount of target miRNA was normalized relative to the amount of U6(ΔCt=ΔCtmiRNA-ΔCtU6).Relative expression levels were expressed as 2 △-ΔCt.Data were analyzed using SPSS 13.0 software.The test of homogeneity of variance and unpaired t-test was used to compare between groups.P values(2-tailed) less than 0.05 were considered as statistically significant.ResultsThe expressions of PBMC and plasma miR-155 were higher in RA patients than those in the healthy control individuals(0.08±0.08 vs 0.05±0.03,t=-2.225,P＜0.05; 5.9±6.7 vs 1.3±2.0,t=-3.677,P＜0.05).The expression of miR-146a in PBMC and plasma of RA patients and controls were(1.3±1.2 vs 0.8±0.6,t=-2.154,P＜0.05)and(741±1001 vs 300±295,t=-1.669,P＞0.05).According to their DAS28 value,RA patients were divided into high activity group (23 cases,DAS28≥5.0) and low disease activity group( 11cases,DAS28＜5.0).The plasma miR-155 and miR-146a expressions were significantly higher in high activity group than those in low activity group.There were no significant differences in the expression of PBMC miR-155 and miR-146a between the two groups.ConclusionThe expression of PBMC and plasma miR-155 and miR-146a are higher in RA patients.The expression of plasma miR-155 and miR-146a are associated with RA patients' activity.Plasma miR-155 and miR-146a may be potential non-invasive biomarkers for RA diagnosis anddisease activity assessment.

Objective To study the effect of subarachnoid hemorrhage (SAH) on voltagedependent potassium channels (Kv) currents of smooth muscle cells,which is hypothesized to be different between cerebral pial arteries and penetrating arteries.Methods Smooth muscle cells from cerebral pial arteries and penetrating arteries in rats were enzymatically isolated 72 h after SAH,and patch clamp was used to test the relative cell size,resting potential and Kv currents.Results Resting potential of either pial ((45.63 ±1.18) mV) or penetrating artery ((41.55-± 1.19) mV) was shifted positively by SAH,even more significantly in latter (F =8.24,P ＜ 0.05 ; F =9.36,P ＜ 0.01) ; Resting potential of pial artery of control ((38.76 ± 1.03) mV),penetrating artery of control ((38.53 ± 0.67) mV),pial artery of SAH ((36.87 ± 1.49) mV) and penetrating artery of SAH((37.89 ± 1.24) mV) were shifted positively to the same level by 1 μmol/L 4-aminopyridine (4AP; F =3.08,P ＞0.05).Maximum Current Density (Imax) of either pial ((20.82 ±0.59) pA/pF) or penetrating artery ((15.15 ±0.37) pA/pF) was compromised by SAH,also more significantly in latter (F =6.22,P ＜ 0.05) ; Imax of pial artery of control (9.15 ± 0.16),penetrating artery of control (9.04 ± 0.36),pial artery of SAH (8.77 ± 0.26) and penetrating artery of SAH (9.12 ± 0.17) were decreased to the same level by 1 μmol/L 4AP (F =2.96,P ＞ 0.05).Conclusions SAH probably shares the similar pathway with Kv blocker (4AP) in Kv currents inhibition.Further,SAH differently inhibits smooth muscle cells Kv currents and resting potential of cerebral pial arteries and penetrating arteries,which may be related with their different sensitivity towards cerebral vasospasm following SAH.

Aim To study the preventive and therapeutic effects of Buyang Huanwu Decoction(BYHWD)on denervated tibial muscle atrophy of rats.Methods After 60 Sprague-Dawley rats were subjected to Common Peroneal nerve crush model of 5 mm injury, they were randomly divided into 6 groups for daily intragastric administration of drugs:BYHWD high-dose, medium-dose, low-dose groups, mecobalamin group(positive control), model group, sham operation group.The drug administration lasted for 18 days.18 days after operation, slice, masson staining and analysis morphology were performed. The wet weight ratio and section area of tibial muscle were also measured.Results Tibial muscle section area of sham operation group was large, morphous was regular, while that of model group significantly diminished, structure was chaotic, and connective tissue hyperplasia was more obvious;Tibial muscle section area of BYHWD group and mecobalamin group was fairly diminished, morphous was fairly regular, and connective tissue hyperplasia was not obvious.Compared with model group, the section area of BYHWD group all increased significantly(P <0.01)in a dose-dependent manner.Compared with mecobalamin group, the cross section area of BYHWD high-dose group was significantly larger(P <0.05).Compared with model group, the wet weight ratio of BYHWD high-dose and medium-dose group all increased significantly(P <0.01 or P <0.05)in a dose-dependent manner.Conclusion BYHWD has a significant effect of prevention and therapy on denervated tibial muscle atrophy of rats.

Objective To study the effect of spontaneous subarachnoid hemorrhage (SAH)on transient receptor potential melastatin 4 (TRPM4)channel activity. Methods Seventeen SD rats of clean grade were selected. They were randomly divided into either a SAH (n = 10)or a sham operation group (n = 7) according to the random number table. At day 5 after SAH modeling,the cerebral arteries were harvested and the cerebral arterial smooth muscle cells were isolated using enzymatic digestion method. Western blot was used to detect TRPM4 expression and translocation rate. Patch-clamp techniques were used to study the maximum current intensity of the TRPM4 single channel in cerebral arterial smooth muscle cells. Results The fluorescent-stained TRPM4 were observed in cerebral arterial smooth muscle cells in the 2 groups of rats. The relative quantities of TRPM4 in the total protein of the sham operation group and the SAH group were 24 ± 3% and 32 ± 4% respectively. There was significant difference between the 2 groups (t = 4. 47,P < 0. 01). The translocation rates of TRPM4 in the sham operation group and the SAH group were 44. 0 ± 1. 9% and 60. 1 ± 2. 3% respectively,and the SAH group was higher than the sham operation group (χ2 = 4. 48,P < 0. 05). When the clamping voltages were - 100 mV,- 80 mV,- 60 mV,and - 40 mV,the maximum current intensity of TRPM4 single channel of the sham operation group was more than that of the SAH group. There were significant differences between the 2 groups (- 1. 90 ± 0. 10 mV vs. - 2. 23 ± 0. 08 mV,- 1. 68 ± 0. 12 mV vs. - 1. 99 ± 0. 12 mV,- 0. 89 ± 0. 09 mV vs. - 1. 24 ± 0. 09 mV,and - 0. 69 ± 0. 12 mV vs. - 0. 92 ± 0. 11 mV;all P < 0. 01). When the clamping voltages were - 20,0,20,40,60,80,and 100 mV,there was no significant difference in the maximum current intensity of TRPM4 single channel between the 2 groups (all P > 0. 05). Conclusion SAH has the induced effect for TRPM4 activity.

Objective To investigate the effect of arterial and venous subarachnoid hemorrhage ( SAH)on voltage-dependent calcium channel( VDCC)currents of cerebral artery smooth muscle cells and the relationship between the concentration of oxyhemoglobin( OxyHb)in arterial and venous blood and cerebral blood flow. Methods Thirty-six clean grade rats were colleted. A rat SAH model was induced by injection of autologous arterial or venous blood in suprasellar cistern using assisted stereotaxic apparatus. The rats were divided into three groups:an arterial SAH( n=14 ),a venous SAH( n=13 ),and a sham operation( n=9 )group. The arterial and venous OxyHb concentrations were measured. Three days after SAH modeling,a patch clamp was used to detect the relative surface area of the cerebral artery smooth muscle cells,resting potential,and VDCC currents in rats. A fluorescent microsphere method was used to quantitatively analyze cerebral blood flow(CBF). Results (1)Arterial SAH OxyHb concentration (127 ± 4 g/L)was significantly higher than venous SAH OxyHb concentration(54 ± 6 g/L),and that of the sham operation group was 50 ± 5 g/L. The differences were statistically significant among the 3 groups( P<0. 01).(2)The maximum current of VDCC of the arterial SAH group(3. 22 ± 0. 31 pA)was significantly higher than that of the venous SAH group(2. 19 ± 0. 27 pA)and the sham operation group(2. 18 ± 0. 29 pA). The differences were statistically significant among the 3 groups( P<0. 01 ). The VDCC currents of the arterial SAH group were consisted of L- and R-currents,while the currents of the venous SAH group were only consisted of L-VDCC.(3)The cerebral blood flow of the arterial SAH group(0. 83 ± 0. 14 mL/[g·min])was significantly higher than that of the venous SAH group(1. 28 ± 0. 28 mL/[g·min])and the sham operation group(1. 35 ± 0. 19 mL/[g·min]). The differences were statistically significant(P<0. 01). Conclusions The changing effect of arterial SAH on the expression and function of the cerebral artery smooth muscle cells are greater than that of the venous SAH. This difference may be associated with the concentration and composition of vasospasm factors of OxyHb in arterial and venous blood.

Objective To investigate the effect of transient receptor potential M4 (TRPM4) on autonomous regulation disorder of cerebral blood flow following subarachnoid hemorrhage (SAH) in rats.Methods A total of 120 clean grade male SD rats were selected.They were divided into sham operation,SAH,negative control,and treatment groups according to the random number table.The dead rats were excluded.A SAH model was induced by using the suprasellar cistern injection method with a stereotaxic apparatus.Isotonic saline 0.2 ml was injected into the rats of the sham operation group and negative control group respectively,and autologous tail arterial blood 0.2 ml was injected into the rats of the SAH group and the treatment group respectively.The isotonic saline solution was continuously pumped into lateral ventricle of rats via implantable micro-pump in the sham operation group and the SAH group respectively,and the concentration of 0.03 mol/L of TRPM4 blocking agent was continuously pumped into the lateral ventricles of rats in the control group and the treatment group respectively.The 4 groups of rats received the regional cerebral blood flow and whole cerebral blood flow detection on day 3,5,and 7,respectively.Results One hundred and six (88.3%) of the 120 SD rats survived to the time point of study,data analyses were performed in the 4 groups (with 21 rats in each group) respectively (n=7 in each time point).There were significant differences in cerebral cortex local and whole cerebral blood flow at day 3,5,and 7 in the sham operation,SAH and negative control groups (all P<0.05).Cerebral cortex local cerebral blood flow (141±18,148±24,and 168±19 PU,respectively at day 3,5,and 7) and whole cerebral blood flow (93±5,85±5,and 85±6 ml/[100 g·min],respectively at day 3,5,and 7 in the SAH group) were decreased significantly compared with the sham operation group (cortex local cerebral blood flow:235±17,220±24,and 224±20 PU),whole cerebral blood flow (141±10,147±8,and 143±8 ml/[100 g·min]),all P<0.05).Cerebral cortex local and whole cerebral blood flow (cortical local cerebral blood flow:183±26,173±26,and 187±15 PU,whole brain:114±10,104±9,and 119±5 ml/(100 g·min) in the treatment group were significantly increased compared with the SAH group (all P<0.05).Conclusion TRPM4 has an obvious effect on improving the autonomous regulation disorder of cerebral blood flow after SAH.

Objective To investigate the expression of miRNA-16 in peripheral blood monouclear cells (PBMC)from systemic lupus erythematosus( SLE)patients. Methods Sixteen SLE patients who meet the diagnostic criteria of SLE revised in 1997 American rheumatology and 12 healthy individuals were selected as our subjects. Their peripheral blood were sampled. Total RNAs were extracted and purified. The level of miRNA-16 was determined by quantitative reverse transcription PCR( qRT-PCR). U6 was used as housekeeping control. The amount of target miRNA was normalized relative to the amount of U6(ΔCt =ΔCt miRNA-ΔCtU6 ). Relative expression levels were expressed as 2-ΔCt . Results The expression level of miRNA-16 in the SLE patients was 919. 87 ± 715. 45,significantly higher than that in the healthy control group(413. 6 3 ± 330. 69;t= -2. 497,P﹤0. 05). And miRNA-16 expression in SLE active group was 1 298. 79 ± 803. 79,significantly higher than that in SLE stable group(540. 95 ± 350. 15;t= -2. 445,P﹤0. 05). The level of miRNA-16 was related with AnuA (r=0. 669,P=0. 005),ESR(r=0. 608,P=0. 012)and SLEDAI(r=0. 530,P=0. 035). Conclusion The expression of miRNA-16 is high in SLE patients and it is related with SLE activity.

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.