Objective To investigate the gene therapy of hypoparathyroidism (HPT) with hemopoietic stem cell s(HSCs) integrated with parathyroid hormone (PTH) gene.Methods Human PTH gene was amplified by polymerase chain reaction from pcDNA 3.1 -PTH templates and inserted into murine stem cell virus (MSCV) vectors. The plasmids recombinated with PTH gene were transfected into PA317 packaging cells and screened for positive clones with Geneticin. The condensed recombinant retroviruses were obtained and used to infect HSCs coming from umbilical cord blood. The infected HSCs were injected into the mice suffering from HPT. The change of symptoms and serum level of PTH and calcium in each group of mice was observed.Results Human PTH gene was proved to be inserted into MSCV vectors and the titres were up to 2?10 7 colony forming unit (CFU)/ml in condensed retroviral solution. The secretion of PTH reached 15 ng per 10 6 cells per 48 h. The wild type viruses were not detected by PCR amplification. The mice suffering from HPT recovered quickly and the serum level of calcium and PTH remained normal for about 3 months after the HSCs recombinated with PTH gene were injected into them. In the groups of mice subject to the injection of the recombinant retroviruses containing PTH gene, the serum level of PTH and Calcium was significantly increased in a short time, and then declined slowly. And the mice took on the manifestation of hypoparathyroidism again. The mice only subject to the implantation of HSCs were all dead within 20 days.Conclusion A long term therapeutic effect could be obtained by infusing the HSCs which were integrated with PTH gene into the mice suffered from hypoparathyroidism.

Objective To explore the pathogeny, pathology, ultrasonic and clinical expression of upper extremity deep venous flow obstruction and superior vena cava syndrome. Methods Retrospective analysis and summarize the disease changes of 10 cases in recent 5 years. Results 7 patients of 10 cases were superior vena cava syndrome, in which 4 cases caused by upper right lung malignant tumor, 3 cases caused by upper and media mediastinal malignant tumor. 2 patients of 10 cases were found thrombosis in right subclavian vein caused by duct of artificial pacemaker. The left 1 patient was no reason thrombosis in right axillary vein. Conclusion Ultrasonography combine with color and spectral Dopper image can evaluate the clinical development and therapeutic efficiency, which is a useful and effective non-invasive examination.

Animals ; Coxsackievirus Infections ; metabolism ; Enterovirus B, Human ; Female ; Male ; Mice ; Myocarditis ; metabolism ; Myocardium ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

4 weeks of arrested embryos.The AAH expression was found to have the similar result as HIF-1?'s.Conclusions The expression level of HIF-1? and AAH in villi of missed abortion patients is much lower than that of normal early pregnant women.HIF-1? and AAH have a function of supporting normal pregnancy,so their low expression may be an important cause of missed abortion.

Objective To investigate factors influencing islet isolation and purification in dogs and to explore how to obtain massive purified viable islets from canine pancreas.To provide experimental evidence for pancreatic islet transplantation in clinical practice.Methods Islets were isolated from pancreas of 22 dogs by modified automated techniques,followed by purification using continuous density gradients.The purity of islets was determined by dithizone (DTZ) staining,and the function of islets was evaluated by glucose stimulating insulin release test.Results The crude islets yield was ( 155 040 ±310) IEQ/pancreas while the yield was (74 200 ±185) IEQ/pancreas after purification.The average purity was (89.4 ±2.6) ％.The recovery rate was(47.8 ±1.3) ％.Islet viability was(93.0 ± 1.7)％.The modified techniques improved both the yield and the quality in islet isolation and purification.The islets were morphologically intact after purification.Glucose stimulating insulin release test showed that the difference of the secreted insulin concentration had statistical significance ( P ＜0.001 ) between low concentration glucose group and high concentration glucose group.Conclusion It is of great value for clinical research to obtain enough islets through islet isolation and purification in dogs.

PurposeTo evaluate the existence and extent of magnetic resonance(MR) artifacts caused by frequently used metallic dental materials and to compare the influence of different MRI sequences on artifacts.MethodsA total of 22 kinds and 25 metallic dental samples were tested with 1.5 T MR imager and gradient-echo sequence. Spin-echo and fast spin-echo were added to parts of these samples. Results Of all the 25 metallic dental samples, 11 including gold, amalgam, and silver point did not produce artifact. Titanium alloy and porcelain product fused in metal had mild artifacts. Whereas the remaining 12 samples such as the retention pin and pivot pin showed severe artifacts. Artifacts produced by retention pin, nickel chromium crown and so on were less severe on fast spin-echo. ConclusionsAttention should be paid to some of the metallic dental materials, which could cause severe MR artifacts and image degradation, when undergoing face,jaw and head MR examination. Artifacts can be alleviated by using proper metallic materials or choosing proper imaging sequence and parameters.

OBJECTIVETo explore the analgesic effect of Zhixin Formula (ZF) and its effects on spinal glial fibrillary acidic protein (glial fibillary acidic protein, GFAP, a marker of astrocyte) , CD11b (a maker of microglia), Toll like receptors (TLR2 and TLR4) and nuclear factor κB (NF-κB) in bone cancer pain model rats.

METHODSTotally 20 male SD rats were randomly divided into the blank control group and the bone cancer pain group, 10 in each group. The bone cancer pain model was induced by injecting ascites tumor fluid containing 3 x 10(3) Walker256 cell line from the left tibia. Ethological tests, X-ray test, and HE staining were performed to confirm a successful modeling. After model was successfully established, 70 male SD rats were randomly divided into seven groups, 10 in each group: the blank control group, the bone cancer pain group (as the model group), the Western medicine (WM) group (Tramadol Hydrochloride), the high dose ZF group, the middle dose ZF group, the low dose ZF group, and the Chinese medicine (CM) group (Wulin Zhitong Capsule). Fourteen days after modeling, rats in the high, middle, and low dose ZF groups were administrated by gastrogavage with 9, 4.5, and 2.25 g/kg ZF water condensed preparation respectively, once a day for seven consecutive days. On day 21 MS typical protein expressions including GFAP, CD11b, TLR (2,4) and NF-κB from cornu dorsal medullae spindis L4-L5 were detected by immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.

RESULTSCompare with the blank control group, increased weight in the model group was slow and showed a decreasing trend (P < 0.01), spontaneous ambulatory pain score (SAPS) obviously increased (P < 0.01), paw withdrawal threshold (PWT) obviously decreased in the model group (P < 0.05, P < 0.01). Results of lateral tibial X-ray and HE staining showed obvious changes and damage occurred in bone structures of rats in the model group. Immunohistochemistry showed that GFAP expression significantly increased in the model group (P < 0.05). Protein levels of NF-κB also significantly increased in the model group (P < 0.05). Compared with the model group, CD11b expressions obviously decreased in the middle and high dose ZF groups (P < 0.01). Meanwhile, protein expressions of TLR2 and TLR4, as well as NF-κB also obviously decreased (P < 0.05).

CONCLUSIONZF had analgesic effect, which might be probably related to inhibiting proliferation and activation of gliocytes, as well as activation of TLRs and NF-κB.

Animals ; Astrocytes ; Bone Neoplasms ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Glial Fibrillary Acidic Protein ; Male ; Microglia ; NF-kappa B ; metabolism ; Osteosarcoma ; Pain ; Rats ; Rats, Sprague-Dawley

Objective To discuss the midterm results of modular femoral prosthesis in total hip revision surgery for bone defects.Methods From December 2001 to June 2006,by using Link-MP modular femoral prosthesis for muhiple reasons(48 with asepsis loosening,seven with infections using two-stage revision procedure,one with fracture of proximal femur and one with periprosthetie fracture), total hip revision surgery was carried out in 56 eases including 24 males and 32 females with age range of 38-77 years(mean age 58.8years).Causes for revision included sterile prothesis loosening in 48 cases, infection of hip prosthesis in seven and peripheral fracture of femoral stem fracture prosthesis in one.Re- vision for infected femur was all at stageⅡ.Of 56 cases with femoral stem prosthesis loosening,30 had loosening of primary cemented prosthesis and 26 of uncemented prosthesis.According to the Mallory bone defect classification,five eases were with typeⅡbone defect,21 with typeⅢA,28 with typeⅢB and two with typeⅢC.Bone grafting was performed in 12 cases and wire or cable cerelage in 28.Fracture of great trochanter was found in two cases,fracture of femoral stem in three and perforation of femoral stem in one.Results A total of 52 eases half year after operation were followed up for mean 31.78 months (8-56 months).No migration of distal femoral stem was found in all eases except for one ease had 1.5 cm subsidence of proximal femoral stem.The Harris hip score was preoperative 46 scores(21-52)and post- operative 89(79-94).There found no significant limb discrepancy,thigh pain or dislocation.Conclu- sions Total hip revision surgery for femoral bone defect using modular femoral prosthesis has optimal midterm result especially in its advantages of regulating limb length,offset,anteversion,which can help us match the proximal femur with distal femur and achieve initial and long-term stability.

Objective To explore the relationship of phosphatase and tensin homology deleted on chromosome ten ( PTEN ) expression with the positioning of AMPA recepter subunit GluR2 in hippocampal neuron after stretch injury. Methods The primary rat hippocampal neuron from neonate rats were cultured and the model of stretch injury established. Western blot and TUNEL staining were used to detect PTEN expression and the positioning of AMPA receptor subunit GluR2 in rat hippocampal neuron at different time intervals after stretch injury. Results The PTEN expression was increased obviously after stretch injury, while the expressions of AMPA receptors GluR1, GluR2 and GluR3 showed no evident differences at every time point. However, AMPA receptor subunit GluR2 located at cytomembrane was down-regulated obviously after stretch injury. Conclusion The intracellular PTEN expression and the expression of AMPA receptor subunit GluR2 located at cytomembrane are on the contrary, indicating that PTEN phosphatase may participate in the transport and reintegration process of AMPA receptor subunit GluR2 located at cytomembrane.

Background Congenital fibrosis of extraocular muscles (CFEOM) affects patient' s appearance and quality of life,and no effective treatment for this disease is available.Imaging study is helpful for exploring the pathogenesis of CFEOM.Objective This study was to describe the characteristics of CFEOM associated with limb movement disorder using magnetic resonance imaging (MRI).Methods A family with CFEOM associated with limb movement disorder was investigated in Renmin Hospital of Wuhan University.Disease history was collected and the pedigree was investigated.Ophthalmologic examinations,including corrected visual acuity,refractive error,slitlamp examination,ophthalmoscopic examination,force of levator palpebrae superioris,ocular movement,eye position,forced duction test,and bell phenomenon examination,were performed.Ocular orbital and cranial MRI was performed in 4 CFEOM patients and 10 normal subjects to compare the structures of the extraocular muscles,motor nerve and cranium.Oral informed consent was obtained from each patient prior to any medical examination.Results A total of 1 1 members from 3 generations were investigated in this study,presenting with 4 cases of disease.The mode of inheritance of this family complied with the Mendelian autosomal dominant inheritance law.Clinical signs included disturbance of eye movement,deviation of eye position,ptosis,lack of Bell sign and positive reaction of passive pull test.In addition,unstable gait,improper body limb alignment,dysphasia and mental retardation were ohserved in 1 patient,which coincided with the diagnostic criteria of type 3 CFEOM.MRI results demonstrated that the levator palpebrae superioris,superior rectus and superior oblique muscle were clearly thinner,and the medial rectus,lateral rectus,inferior rectus muscle were thinning in the patients,showing significant differences in comparison with the normal controls(P＜O.05).The oculomotor and abducens nerves became thinner and even absent in the patients.Cranial MRI showed that Ⅲ-3 in the pedigree with callosum was shorter than that of the normal controls,suggesting that patient suffered from corpus callosum hypoplasia.Meanwhile,cranial MRI indicated the presence of cerebellar hypoplasia and the expansion of the fourth ventricle.Conclusions MRI demonstrates consistent abnormalities of the oculomotor nerves and abducens nerves in the affected individuals in this CFEOM 3 family,and some affected members exhibit two types of central nervous system abnormalities-corpus callosum and cerebellar hypoplasia.These findings suggest that CFEOM 3 is primarily a neuronal disease.