Objective To screen a cell line which stably suppresses DAPK expression and to observe the growth characters.Methods Four pairs of shRNA were designed,synthesized and inserted into the pDsRed1-N1-U6 vector.The recombinant plasmids were purified and transfected into PC12 cell.Meanwhile,a pDsRed1-N1-U6 vector was transfected as control.The cell clones were screened by G418,and the stable PC12 cell line was established.DAPK expression was detected by Western blot.MTT method and Flow Cytometry(FCM) assay were used to assess the growth characters of the cell line.Results The shRNAs were transfected into PC12 cell and the cell clones were successfully screened out.Of the four recombinant plasmids,the F2 was the best interfering shRNA.Beyond our expectation,the F1 recombinant plasmid had an enhanced effect on DAPK expression.Conclusion A stable PC12 cell line with stable inhibition of DAPK expression by was established using siRNA expression vectors.

Objective To analyze the reasons and mechanisms of stereoscopic function abnormality in elder patients after the cataract extraction with intraocular lens implantation. Methods 150 cases with artificial lens implantation were randomly selected.Routine eye examinations were carried out and the associated refractive error and presbyopia were corrected.Bilateral simultaneous visual perception and fusion function were examined.The stereoscopic function was tested using "the stereoscopic function examination diagram"created by Yan Shao-ming.Ninety-seven patients who were found to have unrecover-ed or abnormal stereoscopic function were enrolled for analysis. Results In these 97 cases,53(54.6%)were found to have preexisting eye disorders that could affect visual acuity and binocular single vision before the operation.Macular problem was the most prevalent problem in this group.Twenty-seven(27.8%)patients had complicated with corneal astigmatism,after cataract,paralysing strabismus and diplopia as well as macular edema after the operation.In addition,the contralateral unoperated cataract in 17(17.5%)patients and post-operative anisometropia in 9(9.3%)patients were also the causes of stereoscopic function abnormality.There was no reason could be identified in 8 cases. Conclusions The pre-existing eye disorders before lenses implantation,complications of the operation,contralateral unoperated cataract and anisometropia are all the major factors that affect visual acuity recovery and bilateral stereoscopic function rehabilitation.

Objective To test serum differentially expressed proteins between early-stage (stage IB-ⅢA) and late-stage (stage Ⅳ) lung cancer patients by proteinchip technology and investigate its clinical value. Methods SELDI-TOF-MS and WCX-2 protein chip were used to detect the serum protein of 30 cases of early stage lung cancer patients and 30 cases of late stage lung cancer patients. The data were analyzed by using Biomarker Wizard software. Results There are ten different proteins in the serum between the two groups of lung cancer patients. Four protein markers 7978, 8139, 15 951 and 16 133 are over expressed and seven protein markers 2867, 6885, 8701, 8840, 13 781 and 13 955 are low expressed in the late group. Conclusion SELDI-TOF-MS proteinchip technology is a convenient, sensitive and high-throughput analysis method which can screen several relatively specific protein markers for late stage lung cancer from the serum samples. This selected protein markers can predict metastasis of lung cancer patients.

Objective To study the method and effect of deep inferior epigastric perforator flap(DIEP)in repairing the large defects of lower limbs.Methods Eight cases,from July 2009 to November 2011,including 3 cases of plantar skin defects with bone exposure after foot injuries,three cases of plate exposure after tibia fracture surgery and 2 cases of heel repeated ulceration after skin graft,were repaired by deep inferior epigastric perforator flap.Results All deep inferior epigastric perforator flaps survived with good functions,except 1 case whose distal with poor blood supply and the flap survived after treatmenting,three cases of flap bloated with good appearances after second operation.Conclusion DIEP is a proper option for repair of large defects of lower limbs.It has the advantages of abundant blood supply,large flap area,abdomen can suturing without abdominal complications.

Objective To investigate the role of HMGB1 in the pathogenesis of organ injury of acute necrotizing pancreatitis (ANP).Methods Male ICR mice were randomly allocated into control group,ANP group and HMGB1 monoclonal antibody group.ANP model was induced by intraperitoneal injection of 20％ L-arginine.Mice in HMGB1 monoclonal antibody group were given intraperitoneal injection of 200 μg of HMGB1 monoclonal antibody immediately after the induction of the ANP model.All the mice were sacrificed at 12,24,and 48 h after ANP induction.Serum level of amylase and liver,renal function were determined,level of serum HMGB1 was measured by using enzyme-linked immunosorbent assay,and then the pathologic changes of pancreas and liver were routinely observed and scored.The HMGB1 mRNA levels in the liver and pancreas were studied by real time fluorescence quantitative PCR.Results The serum levels of HMGB1 at 12 h in control group,ANP group and HMGB 1 monoclonal antibody group were (9.09 ± 1.03),(25.04 ± 4.30),(16.84 ± 4.27) μg/L; and pathological scores of pancreatic tissue were (1.50 ± 0.55),(4.33 ± 0.52),(3.03 ± 0.32) points ; and HMGB1 mRNA expressions in pancreas were 0.48 ± 0.18,7.53 ± 2.71,3.26 ±2.33 ; HMGB1 mRNA expressions in liver were-1.23 ± 0.37,0.15 ± 0.65,－ 1.27 ± 0.72.The corresponding values in ANP group were significantly higher than those in control group (P ＜ 0.05).While the corresponding values in HMGB1 monoclonal antibody group were significantly lower than those in ANP group (P ＜0.05).There was a positive linear relationship between serum HMGB1 level and pancreatic pathological scores 24 h after ANP induction (r =0.768,P ＜ 0.05).In addition,the serum levels of AMY,AST,ALT,LDH,BUN,Cr showed a similar trend as that of serum level of HMGB1,and the serum level of HMGB1 was positively associated with serum levels of Cr,BUN and ALT (r =0.824,0.719,0.590,P＜0.05).Conclusions HMGB1 may be a key factor of inflammatory response and organ dysfunction of ANP in mice,and extrinsic supply of its monoclonal antibody may decrease the injuries of pancreas and other organs during ANP.

Objective To investigate the clinical and imaging features in patients with vertebrobasilar dolichoectasia (VBD) through a comparative study in patients with ischemic stroke with or without VBD.Methods The patients with acute cerebral infarction were divided into either a VBD group or a non-VBD group according to magnetic resonance angiography.The VBD group was further divided into an anterior circulation infarction subgroup and a posterior circulation infarction subgroup.The cardiovascular risk factors,the diameter of basiar artery (BA),bifurcation height,and horizontal displacement were compared in all groups.Results A total of 269 patients with acute cerebral infarction were included,28 had VBD,accounting for 10.41％ of the patients with acute cerebral infarction during the same period.The proportion of male patients (78.6％ vs.66.8％ ;x2 =4.392,P =0.036),age (70.38 ± 10.58 years vs.62.86 ± 12.20 years; t =2.870,P =0.009),and the proportion of hypertension (89.3％ vs.47.7％ ; x2 =17.367,P =0.000) in the VBD group were significantly higher than those in the non-VBD group.The multivariate logistic regression analysis showed that the advanced age (odds ratio [OR] 1.248,95％ confidence interval [CI] 1.137-1.371; P=0.000),hyperglycemia (OR 1.599,95％ CI 1.181-2.164; P =0.002),hypertension (OR 1.251,95％ CI 1.020-1.534; P =0.032) and increased triglyceride level (OR 1.876,95％ CT 1.021-3.445; P =0.043) were the independent risk factors for VBD,while female gender (OR 0.133,95％ CI 0.024-0.735; P =0.021) was the independent protective factor for VBD.Of the 28 cerebral infarction patients with VBD,9 had anterior circulation infarction and 19 had posterior circulation infarction.There were significant differences in BA diameter ([5.40 ± 0.49] cm vs.[6.00 ± 0.77] cm; t =2.046,P =0.041),and the proportions of high score in bifurcation height (x2 =6.768,P =0.037) and horizontal displacement (x2 =5.241,P =0.042) between the 2 groups (all P ＜0.05).The multivafiate logistic regression analysis showed that the BA bifurcation height was an independent risk factor for posterior circulation infarction (OR 1.347,95％ CI 1.069-2.457; P =0.038) in patients with VBD.Conclusions VBD accounted for 10.41％ of the patients with acute cerebral infarction during the same period.Advanced age,hyperglycemia,hypertension and increased triglyceride level were the independent risk factors for VBD.Female gender was the independent protective factors for VBD,and the BA bifurcation height was an independent risk factor for VBD occurring posterior circulation infarction.

Objective To investigate the relation ship between the changes of platelet parameters and intracranial hemorrhage in premature children with low birth weight. Methods 73 premature children with low birth weight were selected as research subjects and divided into PIVH group (35 cases) and non PIVH group (38 cases) according to intracranial hemorrhage or not,and 20 normal newborns were selected as controls. The neonatal platelet parameters of three groups were detected and compared. Results There were 35 cases of 73 premature children with low birth weight occurred intracranial hemorrhage, the incidence rate was 47. 9%. The gestational age and birth weight of PTVH group were (29.3 ± 1.2) weeks and (1 653.0 ± 182. 1) g and which were significantly lower than the non PIVH group( all P ＜0.05) ; the asphyxia rate of PIVH group was 60.0% and which was significantly higher than the non PIVH group(P＜0.05). The PLT and PCT of PIVH group were (187.52 ±52.03) × 109/L and (0.127 ± 0.05) % and which were significantly lower than the control group (all P ＜ 0. 05) ; The PLT and PCT of non PIVH group were(223.48 ±42.15) × 109/L and (0. 189 ±0. 06)% and which were significantly lower than the control group(all P＜0. 05) ; The PLT and PCT of PIVH group were significantly lower than the non PIVH group(all P ＜ 0.05) ; while the MPV and PDW among the three groups had no significant difference ( all P ＞ 0. 05 ). Conclusion The abnormal decrease of PLT and PCT should be involved in the pathogenesis of intracranial hemorrhage in premature children with low birth weight, clinical attention should be paid to the monitoring of platelet parameters in premature children so as to alert and minimize its incidence.

BACKGROUND:Mycobacterium tuberculosis heat shock protein 10 (r-Mt cpn10) is one of the main factors that cause bone tuberculosis dissolution and absorption as wel as inhibits the proliferation of osteoblasts. Receptor activator of nuclear factor kappa B ligand and osteoprotegerin are the important factors influencing bone metabolism.
OBJECTIVE:To observe the effect of r-Mt cpn10 on human osteoblast proliferation, alkaline phosphatase secretion, expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA. METHODS:Human bone marrow stromal cel s were induced to differentiate into osteoblasts, and osteoblasts at passage 3 were cultured with various concentrations of r-Mt cpn10 (0.1, 1, 10 mg/L). Osteoblasts cultured without r-Mt CPN10 were assigned as controls.
RESULTS AND CONCLUSION:MTT assay results showed that, compared with control group, r-Mt cpn10 at different concentrations inhibited osteoblast proliferation and alkaline phosphatase secretion (P<0.05). RT-PCR analysis showed that, r-Mt cpn10 at different concentrations increased receptor activator of nuclear factor-kappa B ligand mRNA expression (P<0.01), and inhibited osteoprotegerin mRNA expression in a concentration-dependent manner (P<0.01). 10 mg/L r-Mt cpn10 exhibited the strongest effect (P<0.01). The r-Mt cpn10 can inhibit osteoblast proliferation and alkaline phosphatase activity, and it may influence bone metabolism by regulating the expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA.

Animals ; Basilar Artery ; Brain Ischemia ; pathology ; Brain Stem ; blood supply ; Cats ; Medulla Oblongata ; blood supply

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics