Female ; Humans ; Infant ; Mastocytosis, Systemic ; diagnosis ; pathology ; therapy

Objective To explore the rare primary mt-DNA mutation in LHON patients in China.Design Clinical study.Partici- pants Clinically diagnosed LHON patients whose common primary mt-DNA mutations were negative and normal persons.Methods Polymerase chain reaction(PCR)and DNA sequencing were used to detect the mutations of G11778A,G3460A and T14484C in mt-DNA of clinically diagnosed LHON patients.Fragments of mt-DNA 3962～4356 and 11320～11789 were PCR amplified and se- quenced if all the three mutations above were negative and also in the control group.The sequence results were analyzed by the BLAST service provided by NCBI and compared with the 2001 Revised Cambridge Reference Sequence for the mt-DNA mutations C4171A and G11696A.Main Outcome Measures The sequence results of mt-DNA.Results There were totally 56 eases of clinical diagnosed LHON patients included.The mutation of G11778A was found in 36 cases,G3460A was found in 3 cases,and T14484C was found in 5 eases.Twelve cases of clinical diagnosed LHON patients didn't harbor any of the three common primary mutations,one case harbored the mutation C4171A,the mutation of G11696A was not found.There were totally 25 cases in the control group,the mutations of G11696A and C4171A were not found.Conclusion C4171A may be rare primary mt-DNA mutation in LHON patients in China.(Oph- thalmol CHN,2007,16:382-385)

Objective To explore the guidance significance of the functional MRI and DTI (fMRI ,DTI) ,intraoperative ultra-sound(IOUS) ,neuronavigation ,electrocorticography(EcoG) monitoring used in surgical treatment of low-grade gliomas with epi-lepsy as main symptom located near the eloquent brain regions .Methods 23 neurosurgical patients in the First Affiliated Hospital of Chongqing Medical University during 2009-2010 were performed the retrospective analysis .The preoperative fMRI ,DTI deter-mined the positional relation between the lesions with the conduction bundle and the eloquent brain regions ,the electrophysiological and imageological examinations positioned the epileptogenic focus and lesions ,the MRI-mediated neuronavigation system was adopt-ed to formulate the surgical plan and choose the best surgical approach ,IOUS was used to perform the realtime monitoring for pre-cisely positioning the lesion range and determining the extent of resection ,and the intraoperative EcoG was adopted to determine the epileptogenic focus localization ,the lesions and the epileptogenic focus was dealed by the operating microscope for avoiding the func-tional region ,and the patient′s prognosis was evaluated and recored in detail after operation .Results By the precisely positioning the lesions and epileptogenic focus by fMRI ,DTI ,neuronavigation and ultrasound ,the lesion resection degrees by the operative mi-croscope and intraopertaive pathological guidance were 17 cases of Ⅰ-Ⅱ grade ,4 cases of Ⅲ grade and 2 case of Ⅳ-Ⅴ grade .1 case of motor aphasia ,4 cases of hemiplegia and monoplegia and 1 case of disturbance of consciousness after operation were improved by the treatment of neurotrophy ,dehydration and hyperbaric oxygen and discharged from hospital with rehabilitation .No death case occurred .The evaluation of the life quality :20 cases ofⅠ-Ⅱ grade ,3 cases of Ⅲ grade and no vegetable survival case of Ⅳ grade . The evaluation of resection clinical effect :20 cases of cure ,3 cases of improvement ,no case of as before and exacerbation .After fol-lowed up for 6-24 months ,according to Engel classification of seizure efficacy assessment :Ⅰ-Ⅱ grade in 21 cases ,Ⅲ grade in 2 case ,no case of Ⅳgrade .Conclusion fMRI ,DTI ,neuronavigation ,IOUS and EcoG for guiding the operation of low grade gliomas located near the eloquent brain regions can resect the lesion to the largest extent and simultaneously deal with epileptogenic focus , effectively protect the neurological function of the functional region ,greatly reduce the side-injury of the normal brain tissues in the functional region ,at the same time increase the curative effect of symptomatic epilepsy .

BACKGROUND: Bone morphogenetic protein-2(BMP-2) production of targeted cells is promoted by transfection of adenoviral vectors containing gene, but there are some immune responses. Transfection with plasmid as vector holds promise.OBJECTIVE: To explore the feasibility to construct human bone morphogenetic protein-2 eukaryotic expression vector labeled with green fluorescent protein (GFP).DESIGN: Single sample observation.SETTING: Tianjin Hospital.MATERIALS: The experiment was performed at the Key Laboratory of Hormone and Development, Ministry of Health, Tianjin Medical University from March 2006 to March 2007. pcDNA3.1/CT-hBMP2 plasmid containing full-length hBMP2 gene fragment was provided by Dr. Li; bicistronic eukaryotic expression vector pSELECT-GFPzeo-MCS and Zeo was provided by Invivogen; pTA2(R)-T Easy by Dingguo, China; restriction enzymes BamHI and NheI, T4 DNA ligase by Jingmei Biotech; PCR upstream and downstream primer synthesis and sequencing by Augct, Beijing.METHODS: With pcDNA3.1/CT-hBMP2 as template, hBMP2 target fragment was subcloned by PCR binding with designed specific primers. The fragment was bound with pTA2-T-easy and pSELECT-GFPzeo-MCS, separately, and transfected into DH5 α cells. pSELECT-GFPzeo-hBMP2 containing GFP was obtained after screening.MAIN OUTCOME MEASURES: hBMP2 sequence was identified by PCR; whether hBMP2 was cloned into pTA2-hBMP2 and pSELECT-GFPzeo-MCS was identified by digestion and sequencing.RESULTS: A target fragment of 1 216 bp was obtained by PCR amplification, and cloned into pTA2-T-easy and pSELECT-GFPZeo-MCS. The screening and sequencing results showed that the target fragment was 100% matched with BMP2cDNA sequence (NM-001200) from GenBank.CONCLUSION: hBMP2 eukaryotic expression vector labeled with green fluorescent protein is successfully constructed.

Objective To investigate the expression of insulin-like growth factor 2 receptor(IGF-2R)induced by hypoxia and serum deprivation(H/SD)in cultured neonatal rat cardiomyocytes in vitro.Methods Cultured neonatal rat cardiomyocytes were randomly divided into control group and H/SD group.Expression of IGF-2R mRNA was measured by RT-PCR and real time RT-PCR.Level of IGF-2R protein was measured by enzyme-linked immunosorbent assay(ELISA).Results Compared with control group,the expression of IGF-2R mRNA increased significantly (P ＜0.01)in cardiomyocytes cultured in H/SD condition for 12 h and 24 h,and IGF-2R protein was upregulated in cardiomyocytes cultured in H/SD condition for 24 h(P ＜0.05).Conclusion H/SD may induce IGF-2R expression,and IGF-2R may potentially explain the mechanism of cardiomyocyte damage induced by ischemia.

Objective To investigate the effect of neurolytic celiac plexus block (NCPB) on the inflammatory reaction of the remaining liver tissue and liver function after partial hepatectomy (PH) in rats.Methods Thirty male Sprague-Dawley rats of SPF were constructed as a PH model with deligation and ablation operated on their left and middle lobes,respectively.Then,they were randomly divided into two groups:NCPB group and control group.Twelve hours after the surgery,0.5％ lidocaine was given in the NCPB group once a day,while 0.9％ saline was given in the control group.Determination of liver function,generation of the remaining liver,and deposition of IL-β,TNF-α of the pathological section was respectively made on Day 1,3 and 7 after the surgery.Results On Day 1,3 and 7 after surgery,both the aspartate aminotransferase (AST) and the alanine aminotransferase (ALT) levels in NCPB group were,to different degrees,lower than those in the control group (P ＜ 0.05).Strikingly,total bilirubin in NCPB group was lower than that in control group (P ＜ 0.01) on Day 7,while the level of semm albumin was higher than that in control group (P ＜ 0.01).There was no statistically significant difference on the generation of the remaining livers between NCPB and control groups.On Day 3 and 7,the deposition of IL-β,TNF-α in the pathological sections of NCPB group were lower than those in control group.Conclusion NCPB can not only effectively reduce the damage of liver function caused by PH surgery,but also improve the inflammatory reaction of the residual liver.

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

Objective To investigate the correlation between serum insulin-like growth factor-1(IGF-1)level and the 131Ⅰahsorbante of thyroid nodule in patients with struma nadosa,to search for simpler and safer methods for differentiating thyroid nodule.Methods Detecting the 131Ⅰ absorbance of thyroid nodule by radioisotope scanning.then the patients were divided into warm and cold nodule groups,and the normal control group was also set up;the levels of IGF-1,FT3,FT4,sTSH were detected in serum of patients with struma nadosa by radio immunoassay,then the correlation between these data and the 131Ⅰabsorbance of thyroid nodule was analyzed.Results In the patients with warnl nodule,the level of serum IGF-1,FT3,FT4 and the 131Ⅰ absorbance of thyroid nodule[(315.86±22.74)μg/L,(9.95±5.62),(67.27±27.31)ng/L,0.64±0.17]were increased obviously when compared with the control group [(256.13±39.85)μg/L,(2.80±1.30),(13.51±5.50)ng/L,0.35±0.15],but the sTSH[(0.35±0.03)mU/L]went down significantly than the control group[(2.71±1.17)mU/L],the difference being statistically significant(P＜0.01).In the patients with cold nodule,the level of serum IGF-1,FT3,FT4,sTSH[(263.17±30.23)μg/L,(2.89±0.98),(14.23±2.84)ng/L,(2.81±0.42)mU/L] had no significant difference compared with the control group(P＞0.05).The level of serum IGF-1 was positively correlated with the 131Ⅰ absorbance of thyroid nodule(r＝0.835,P＜0.01),but negtively correlated with sTSH(r＝-0.326,P＜0.05)in the patients with warm nodule.Conclusion The level of sernm IGF-1 is closely correlated with the 131Ⅰ absorbance of thyroid nodule in patients with struma nadosa.

Objective To observe the clinical effect of the club exercise designed ourselves on the prevention of should-hand syndromeafter stroke. Methods 100 stroke patients in early stage, whose motor function of upper limb were lower than Grade Ⅲ evaluated byBrunnstrom assessment and without shoulder pain and swelling of hand, were randomly divided into control group (n=50) and observationgroup (n=50). All of them received routine rehabilitation therapy, and the observation group received club exercise in addition. The incidenceof shoulder pain, the active range of motion of shoulder, the motor function of upper limb and activities of daily living (ADL) of 2groups were evaluated before and 2 courses (60 d) after treatment. Results There was no statistical difference between 2 groups before thetreatment (P>0.05). After 2-course treatment, the incidence of shoulder pain was significantly lower in the observation group than in the controlgroup (P<0.001), the active range of motion of shoulder, motor function of upper limb and ADL were significantly better in the observationgroup than in the control group (P<0.001). Conclusion The club exercise can prevent the pain of shoulder joint, increase the activerange of motion of shoulder joint and improve the motor function of upper limb and ADL in patients after stroke.

BACKGROUND:The physical properties and chemical composition of teeth are very similar with human bone tissue, and there are a larger proportion of inorganic components. Therefore, tooth can be considered as a potential repair material for autologous or alogeneic bone defects. OBJECTIVE:To prepare polyetheretherketone/odontogenic biphasic bioceramic composite, and to test its mechanical properties. METHODS: The humanin vitro teeth of clinical waste were colected. The organic components were removed after a preliminary calcination. Another calcination was conducted after soaking in diammonium phosphate solution for 24 hours to prepare the biphasic ceramics with the main components of hydroxyapatite and β-tricalcium phosphate. The biphasic ceramics were ground and sieved using 200-mesh sieve folowed by impregnation in organic foam to prepare polyetheretherketone/odontogenic biphasic bioceramics. Phase analysis, scanning electron microscopy, elemental analysis, porosity, compressive strength and bond strength test were conducted. RESULTS AND CONCLUSION:Polyetheretherketone/odontogenic biphasic bioceramics presented porous network structure and interconnected holes, with the aperture of 100-800 μm, porosity of 73.65%, compressive strength of (165.260±11.703) N, bond strength of (14.63±6.21) MPa. P element content accounted for 19.8%, and Ca element content accounted for 40.5%. The main phases were β- tricalcium phosphate and hydroxyapatite. These results demonstrate that polyetheretherketone/odontogenic biphasic bioceramics have good mechanical properties.