Objective:Some antigens of M.tb to culture with peripheral blood mononuclear cells ( PBMC) for assaying their proliferation and activation,so as to signify whether lipid antigens of M.tb have specific immune responses in host against M.tb infection or not.Methods:We treated PBMC with several lipid antigens of M.tb to explore the ability of these antigens to activate immunity in healthy individuals.We measured and analyzed cell proliferation by labeling cells with carboxyfluorescein succinimidyl amino ester (CFSE) and subjecting them to flow cytometry (FCM).The production of IFN-γ,TNF-αand IL-4 by T cell subsets (NKT,CD4+, CD8+,andγδT) from healthy donors was analyzed by FCM after stimulation with autologous immature dendritic cells pre-cultured with M.tb lipid antigens.The tested M.tb lipid antigens were the total lipid (TLIP),Acetone-Soluble Lipids (ASLIP),Purified Sulfolipid (PSLIP),Lipoarabinomannan (LAM) and Lipomannan (LM) levels.Medium free of lipid antigens(WCL,CFP,LPS,Mtb-HAg and blank) was used as a control.Results:We found the proportion of proliferative NKT and CD8+T cells significantly increased in all lipid groups (P<0.05).ASLIP,LAM and LM promoted non-proliferative CD4+T cells to secrete IL-4 and proliferative ones to secrete IFN-γ( P<0.05).All lipid antigens promoted both proliferativeγδT cells and CD8+T cells to secrete IFN-γand TNF-α,but the proportion of TNF-α-secreting cells in these populations decreased in the LM group ( P<0.05 ).Conclusion: Lipid antigens may affect the CD1-restricted T cells of the host to fight M.tb infection.

OBJECTIVETo research the clinical application of lower cervical pedicle screw fixation procedure.

METHODSFrom September 2011 to July 2013,32 patients underwent posterior pedicle screw-rod system fixation were retrospective analyzed includinig 20 males and 12 females with an average age of 56.4 years old ranging from 21 to 78 years. Among them, 10 patients were traumatic cervical spinal injury, 9 patients were cervical spinal canal tumors, 7 cases were posterior longitudinal ligament ossification of cervical vertebrae, 6 cases were multiple segmental cervical spondylopathy. Preoperatively, X-ray, computed tomography, magnetic resonance imaging and magnetic resonance angiography of the vertebral artery were performed in all patients. After the operation and during the follow-up,X-ray and computed tomography were performed to confirm the pedicle screw position. The accuracy of the pedicle screw placement was evaluated by 4 grades classification from Lee. The spinal cord function was assessed by ASIA impairment scale for traumatic patients and JOA score for non traumatic patients.

RESULTSTotally 144 pedicle screws performed on 32 patients from C3 to C7 involving 132 screws of grade 0,5 screws of grade 1,5 of screws grade 2 and 2 screws of grade 3 according to postoperative CT. There were 12 screws penetrating the pedicle cortex including 8 screws at lateral,2 screws at caudal, 1 screw at medial and 1 screw at cranial. The follow-up time was 12 to 33 months with an average of (21.0±1.5) months. The spinal cord function was not improved in 6 complete cervical spinal cord injury patients,but their paraplegic level descended 1 to 3 segments. Four incomplete cervical spinal cord injury patients' ASIA impairment scale was increased by 1 to 2 grades in average. The JOA score of 22 atraumatic patients increased from preoperative 11.5±0.8 to 15.9±0.6 of postoperative at 6 months (P<0.01). There were no screw loosening,screw pullout and screw-rod breakage.

CONCLUSIONThe lower cervical pedicle screw fixation can provide excellent 3D stability of the vertebral column. The operation risk and Complication could be minimized by adequate preoperative evaluation for appropriate cases and individual pedicle screw placement. It deserved the clinical expansion.

Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Female ; Humans ; Male ; Middle Aged ; Pedicle Screws ; Retrospective Studies ; Spinal Cord Injuries ; physiopathology ; surgery

No-reflow phenomenon after reperfusion treatment of acute myocardial infarction (AMI) is becoming more recognized today. The effective treatment for no-reflow has not been reported.

Objective To explore the significance of surgical resection for the pineal region tumor,the extent of tumor resection,the surgical approaches and treatment measures of hydrocephalus.Methods Twentythree patients diagnosed of pineal region cancer were recruited for this study.Thirteen patients received ventriculo-peritoneal shunt(V-P),during which 7 cases received intraoperative end-plate colostomy.Two of the 13 cases received a second V-P procedure.Another 13 cases received operation by corpus callosum- septum-Dome Room-the third ventricle approach to remove the tumor; Eight cases underwent the tumor resection by suboccipital supratentorial approach (Poppen approach )and 2 cases underwent the tumor resection by the infratentorial cerebellar approach( Krause approach).Results Total tumor resection was performed in 11 cases,subtotal or major total resection in 8 cases and partial resection in 4 cases.Nine cases underwent postoperative radiotherapy alone,8 receiving radiotherapy plus chemotherapy,and 6 cases receiving neither radiotherapy nor chemotherapy.Two cases died during treatment.Conclusion Though it is not practicable to completely remove the germ cell tumor in patients with relatively large tumors,cerebrospinal fluid circulation pathways should be reestablished,including ventriculo-peritoneal shunt,colostomy from the end of the third ventricle to the end plate.If tumors are not too large,they would be easy to be removed.The effects on the surrounding brain tissue would not be much significant.If the connection of Ⅲ,V ventricles were normal,the patient may not need shunt or colostomy.Radiotherapy and chemotherapy can be given to this kind of patients after the surgery.To improve the efficacy,other types of tumors,expecially the benign tumors,should be totally removed.

Objective:To design and construct the replication-deficient recombinant adenovirus Ad-siCTGF which can silence the expression of connective tissue growth factor (CTGF) by RNA interference and verified its function. Methods:A specific sequence, which was verified to be able to silence CTGF gene with high efficiency, was cloned into pSES-HUS vector to produce the shuttle plasmid pSES-siCTGF. The plasmid after Pme Ⅰ linearization was cotransduced with pAdEasy into BJ5183 E.coli strains to construct recombinant vector Ad-siCTGF. After linearization treatment with Pac Ⅰ enzyme digestion Ad-siCTGF was transfected into HEK293 cells via liposome mediation. The recombinant adenovirus was packaged. The titer of the Ad-siCTGF was increased after three times of cross-infection. 4T1 cells were infected with the adenovirus. The silencing efficiency was tested by real-time fluorescence quantitative (RFQ)-PCR and Western blotting.Results:Pac Ⅰ enzyme digestion electrophoresis indentified that recombinant adenovirus was successfully constructed. The titer of the recombinant adenovirus Ad-siCTGF was 2.6×10~(10) pfu/mL after amplification and purification. The CTGF mRNA and protein expression in 4T1 cells were decreased by 36.27% and 31.56%, respectively, compared with the control groups.Conclusion:The recombinant adenovirus which can silence the expression of CTGF was successfully constructed. It laid a good foundation for further investigation of the action mechanism of CTGF in tumor cells.

Objective To study the relationship between P16,CyclinD1 and P53 anti-oncogene and the gen- esis of hydatidiform mole.Methods 30 samples of hydatidiform moles and normal early pregnant aborted placenta villi respectively were obtained to detect the P16,CyclinD1 and P53 anti-oncogene expression in two kinds of tissues by using SP immunohistochemical staining.Results Compared with that of normal villi,the expressions of P16,P53 and CyclinD1 anti-oncogene were quite different in hydatidiform moles.The expression of P16 was all positive,while CyclinD1 and P53 were all negative in the chorion of early gestation.A descending tendency of P16 expression was found,while the expression of CyclinD1 showed an ascending tendency.The positive rate of P16,CyclinD1 and P53 expression was significantly different between the groups.It was also observed that there was significant difference between the P16 and the proliferation trophocyte.Conclusion P16,CyclinD1 and P53 anti-oncogene have a close relationship with the genesis of human hydatidiform mole.

Objective To explore the diagnostic value of two-dimensional speckle tracking technology in the evaluation of right ventricular systolic function in patients with pulmonary disease .Methods Thirty patients with pulmonary heart disease were divided into two groups:group of compensated pulmonary heart disease(compensated group ) (n =15),group of decompensated pulmonary heart disease (decompensated group)( n =15).30 healthy subjects were enrolled in control group .The displacement of the tricuspid annulus at the midpoint(TADmid),the displacement of the tricuspid annulus at the free wall (TADfre) and the displacement of the tricuspid annulus at the septum (TADsep) were acquired,and simultaneous real-time three-dimensional ultrasound detection of right ventricular ejection fraction (RVEF) were taken.The correlation of TADmid with RVEF and pulmonary artery systolic pressure ( PASP) were analyzed.Results TADmid of the healthy control group,the compensated group and the decompensated group were(17.1 ±3.9)mm, (13.6 ±2.6)mm,and(9.5 ±3.2)mm respectively.TADfre were(21.1 ±3.0)mm,(17.6 ±4.2)mm,and (11.5 ±3.8) mm respectively.TADsep were(12.0 ±2.5) mm,(9.7 ±3.3) mm,and(7.4 ±2.7) mm respectively.RVEF were(56.3 ±8.2)%,(39.6 ±6.4)%,and(28.1 ±5.9)% respectively.PASP were (20.6 ±2.6) mm Hg (1 mm Hg =0.133 kPa), (63.3 ±5.6) mm Hg, and (82.5 ±11.2)mm Hg respectively.There were significant differences of TADmid , RVEF, and PASP among the 3 groups ( F =8.581,7.816,9.300,6.507,10.235, all P <0.05).TADfre, TADmid, TADsep and RVEF were all decreased in the compensated group comparing to the healthy control group ,while PASP was increased.The decrease of TADmid was the most significant ,while that of TADfre was the slightest .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.703,2.536,2.379, 2.817,3.026,all P<0.05).TADfre,TADmid,TADsep and RVEF of decompensated group reduced more significantly than the compensated group , while PASP was increased significantly .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.519,2.493,2.236, 2.621,2.985,all P<0.05);TADfre and TADmid were decreased more apparently in decompensated group than that in compensated group ,and so were TADsep and RVEF,while PASP were increased.There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups (t =1.947, 2.680,2.016,2.653,2.893,all P<0.05).There was significant positive correlation between TADmid and RVEF measured by real-time three-dimensional ultrasound (r =0.904,P <0.01 ).There was significant negative correlation between TADmid and PASP (r=-0.686,P<0.01).The cut-off point value of TADmid measured by speckle tracking technology for evaluation of RVEF <45% and<30% were 13.65 mm and 9.80 mm,respectively.The sensitivity were 94.4%and 90.0%respectively,and the specificity were 78.6%and 90.0%respectively.Conclusions TADmid is hardly affected by the external factors ,and it can better reflect the changes of right ventricular systolic function for patients with pulmonary heart disease .TADmid is positively correlated with RVEF measured by real-time three-dimensional ultrasound and negatively correlated with PASP.Further more,the correlations is significant.The three parameters can authenticate mutually ,and the combination of them can evaluate the right ventricular systolic function in patients with pulmonary heart disease precisely.

Objective To explore the association between polymorphisms of adiponectin gene and the risk of ischemic stroke in Han population from the Northern parts of China.Methods TaqMan probe of RT-PCR was applied to detect the genotype frequency of single nucleotide polymorphism(SNPs)(rs266729 and rs2241766)of adiponectin gene in 357 ischemic stroke cases who developed the episode at first time and with 345 healthy controls.Logistic regression analysis was used to evaluate the relationship of each genotype of SNPs and ischemic stroke.Results Mutation of rs2241766(T＞G)increased the risk of ischemic stroke among all the samples(0R=1.55,P=0.01)and it was still the risk factor of ischemic stroke when analyzed by multi-factors logistic regression after each factor was adjusted(OR=1.55,P=0.00).The polymorphism of rs266729 was not related to the risk of ischemic stroke among all the samples(OR=1.13,P=0.57).However,the genotype GG of rs266729 increased the risk of ischemic stroke among female population(OR=3.25,P=0.04).Conclusion The variance of rs2241766 in adiponectin gene was related to the risk of ischemic stroke in Han population from the Northern parts of China and the genotype GG of rs266729 could possibly increase the risk of ischemic stroke in women of Han population from the Northern parts of the country.

OBJECTIVETo study the subcellular localization of severe fever with thrombocytopenia syndrome virus (SFTSV) in macrophages and understand the replication and assembly mechanism of SFTSV in host cells.

METHODSUsing two types of human macrophage cell lines THP-1 and U937, the study analyzed the intracellular colocalization of SFTSV with Golgi apparatus and endoplasmic reticulum by immunefluorescence staining and confocal microscopy.

RESULTSSFTSV infected macrophage cell lines THP-1 and U937. Immunofluorescence staining showed that the SFTSV nuclear protein colocalized with Golgi apparatus and closely surrounded by endoplasmic reticulum in the perinuclear region.

CONCLUSIONThe results suggested that Golgi complex and endoplasmic reticulum are probably the sites for formation and maturation of SFTSV viral particles.

Bunyaviridae ; isolation & purification ; Cell Line, Tumor ; Endoplasmic Reticulum ; virology ; Fever ; virology ; Golgi Apparatus ; virology ; Humans ; Macrophages ; virology ; Thrombocytopenia ; virology

OBJECTIVETo observe amplitude changes of low frequency fluctuation in brain spontaneous nervous activities induced by needling at Hand Taiyin Lung Channel, and to preliminarily explore the possible brain function network of Hand Taiyin Lung Channel.

METHODSBy using functional magnetic resonance imaging (fMRI), 16 healthy volunteers underwent resting-state scanning (R1) and scanning with retained acupuncture at Hand Taiyin Lung Channel (acupuncture, AP). Data of fMRI collected were statistically calculated using amplitude of low frequency fluctuations (ALFF).

RESULTSUnder R1 significantly enhanced ALFF occurred in right precuneus, left inferior parietal lobule, bilateral superior temporal gyrus, bilateral middle frontal gyrus, left superior frontal gyrus, left inferior frontal gyrus, left medial frontal gyrus. Under AP significantly enhanced ALFF occurred in right precuneus, bilateral superior frontal gyrus, cerebellum, bilateral middle frontal gyrus, right medial frontal gyrus, and so on. Compared with R1, needing at Hand Taiyin Lung Channel could significantly enhance ALFF in right gyrus subcallosum and right inferior frontal gyrus. Significant decreased ALFF appeared in right postcentral gyrus, left precuneus, left superior temporal gyrus, left middle temporal gyrus, and so on.

CONCLUSIONNeeding at Hand Taiyin Lung Channel could significantly change fixed activities of cerebral cortex, especially in right subcallosal gyrus, right inferior frontal gyrus, and so on.

Acupuncture Therapy ; Brain ; physiology ; Brain Mapping ; Humans ; Magnetic Resonance Imaging