Objective In order to observe the role of genetically modified Schwann cell (SC) with pSVPoMcat in the regeneration of injured spinal cord.Method The cells were implanted into the spinal cord.Ninety SD rats were used to establish a model of hemi- transection of spinal cord at the level of T8,and were divided into three groups,randomly, that is,pSVPoMcat modified SC implantation(Group A), SC implantation(Group B),and without cell implantation as control(Group C).After three months the presence of axonal regeneration of the injured spinal cord was examined by means of horseradish peroxidase(HRP)retrograde labeling technique and stereography.Result The results indicated that HRP labeled cells in Group A and B could be found in the superior region of injured spinal cord and the brain stem such as the red nuclei and oculomotor nuclei. The density of ventral horn neurons of the spinal cord and the number of myelinated axons in 100 μ m of the white matter was A >B >C group.Conclusion In brief,the pSVPoMcat modified SC intraspinal implantation could promote regeneration of the injured spinal cord.

Objective To identify the GNAS gene mutation resulting in pseudohypoparathyroidism type Ⅰa (PHP-Ⅰa) in one patient. Methods The clinical data of a patient with pseudohypoparathyroidism type Ⅰa was retrospectively analyzed. All the 13 exons of GNAS were sequenced using Sanger method for the patient and the parents. The distribution of suspected causal mutation was screened in 478 healthy controls. To clarify the origin of the mutation, we performed targeted high-depth sequencing of GNAS exon harboring the mutation for the patient and the parents. Results The clinical data of the patient with the laboratory results of hypocalcaemia, hyperphosphataemia, elevated serum PTH, together with the features of AHO, conformed to the characterization of PHP-Ⅰa. The sequencing of GNAS exons identified a missense mutation (c.479G>C, p.R160P) located at exon 6 in the patient, which was absent in DNA of the parents. The mutation was not reported previously and was not found in the 478 healthy controls. We obtained about 8000-fold coverage from high-depth sequencing of DNA from peripheral blood of the patient and the parents. The disease-associated allele C identified in the patient was not observed in the parents. The number of reads with G allele (3984 reads) was roughly equal to that of C allele (4019 reads) from the targeted reanalysis of DNA of the patient. The results from high-depth sequencing indicated a de novo mutation in maternal germ cells. Conclusions We identified a new GNAS gene mutation (c.479G>C, p.R160P) caused PHP-Ia in a patient. Our results suggested the mutation was a maternal germline de novo mutation.

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

Objective:　To evaluate the effect and safety of early moderate hypothermia therapy (MHT) on patients with head injury by using parallel-control test. 　　Methods:　Thirty patients with severe head injury within 6 hours after accidents were treated by MHT generally for 4-10 days. The other 30 patients with similar head injury treated routinely were enrolled for a parallel-control test. The mortality, morbidity and changes of some neuro-functional indexes as Glasgow Coma Scores, and Glasgow Outcome Scale, levels of endothelin and some other factors of neurobiochemistry in blood plasma were observed. Meanwhile, the dynamic monitoring by transcranial Doppler ultrasonography was conducted in these patients. 　　Results:　The mortality in MHT group was significantly lower than that in control group. MHT not only reduced endothelin but also increased the brain biochemical factors, which were helpful to the protection of neurons in the early brain ischemia after head injury. 　　Conclusions:　Early MHT can help reduce mortality and morbidity in patients with acute head injury.

Objective: To investigate the potential role of me latonin in the adrenal cortex of human embryo. Methods:Specifi c melatonin receptors was localized and characterized in the adrenal cortex of h u man embryo by means of immunohistochemistry. Results: mt1 （Me l1a）and MT2 （Mel1b）subtype of melatonin receptors was principally localize d to cytoplasm in zona glomerulosa， fasciculata and reticularis. Conclu sion: It is possible that mt1 and MT2 subtype of melatonin receptors co-exist in the adrenal cortex of human embryo.

AIMTo investigate the effects of RLI on plasma nitric oxide (NO) and NO synthase (NOS) isoforms of healthy humans.

METHODS30 healthy human subjects (aged from 40 - 70 years old) were recruited. RLI was induced by five 5 min cycles of ischemia of non dominant arm (200 mmHg, 5 min interval). Blood pressure, heart rate, and the feelings of ischemic arm were continuously monitored. Venous plasma was collected in contralateral arm at Pre, Post-0 h, Post-4 h, and Post-24 h. Plasma level of NO was measured by Griess reaction, and NOS was measured by chemical method.

RESULTSBlood pressure and heart rate varied in normal range. The uncomfortable feeling was decreased with the increasing numbers of ischemic cycles. Plasma level of NO, and iNOS in plasma were significantly increased at Post-0 h, Post-4 h, and Post-24 h compared to Pre (P < 0.05). tNOS was also significantly increased at Post-0 h and Post-4 h compared to Pre (P < 0.05). No significant change in plasma cNOS was shown at following three time points than Pre.

CONCLUSIONThese findings suggest that RLI can elevate plasma level of NO, tNOS, and iNOS in healthy humans. RLI might be a safe method as a rIPC, and it would have important possibility to be performed in clinic.

Adult ; Aged ; Arm ; blood supply ; Female ; Humans ; Ischemia ; blood ; physiopathology ; Ischemic Preconditioning ; methods ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; metabolism ; Reperfusion Injury ; physiopathology ; prevention & control

OBJECTIVETo evaluate the efficacy and safty of photoselective vaporization of prostate (PVP) in the treatment of benign prostatic hyperplasia with obstruction within 5 years.

METHODSFrom December 2004 to December 2009, there were 782 cases have been except for neurogenic bladder dysfunction and prostate cancer, who received PVP surgical treatment of BPH. The surgical conditions and postoperative follow-up data were recorded and the follow-up cut-off time for surgery after 5 years.

RESULTSA total of 782 patients with BPH who underwent PVP were included in this retrospective study. The operation in 740 cases was successfully completed at one time. But in other 42 cases, the twice operation was performed. The mean operation time was (85 ± 38) minutes, and the mean energy delivery was (355 ± 124) kJ. The mean catheterization and postoperative hospitalization time was (2.3 ± 1.7) days and (5.2 ± 2.6) days, respectively. No severe intraoperative complications were observed. The mean follow-up was (44.1 ± 19.3) months. The shortest follow-up was 6 months. The longest follow-up was 5 years. Complete follow-up data were available for 398 of the 782 patients. Of the 398 patients followed up for 5 years, the mean international prostate symptom score after 5 years was 12.8 ± 6.9, quality of life score was 2.2 ± 1.6, maximal flow rate was (14.5 ± 2.4) ml/s, and residual urine volume was 58 ml (M50). The retreatment rate because of BPH was 2.3% (9/398). Urethral stricture and bladder neck contracture were observed in 1.5% and 0.5% of the patients, respectively.

CONCLUSIONSPVP has demonstrated remarkably consistent results for objective and subjective voiding parameters. Its late complication is rare and retreatment rate is low.

Aged ; Follow-Up Studies ; Humans ; Lasers, Solid-State ; Male ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome

OBJECTIVETo study the mechanisms of dystrophin gene deletion by cloning and sequencing the junction fragment of dystrophin gene with exons 3 to 5 deletion.

METHODSPCR was performed to verify dystrophin gene exons 3 to 5 deletion in a patient with Duchenne muscular dystrophy. A PCR-based genome-walking method was used to localize the breakpoint in introns 2 and 5, and the deletion-junction fragment was directly amplified by PCR approach with forward and reverse primers annealing to a DNA sequence as close as possible to the breakpoint in the introns 2 and 5. The sequencing result of the deletion-junction fragment was compared with the normal intron sequences.

RESULTSA sequence of 2113 bp containing the junction fragment was obtained. The 5' breakpoint was located in SINE/Alu element of intron 2, and the 3' breakpoint was located in the unique sequence near the sequence TTTAAA. The breakpoints were associated with a strong topoisomerase II cleavage site. A 26-bp fragment was inserted into the breakpoint and formed 3 duplications (GGCTTATATTTAA) of 13 bp around the deletion-junction fragment.

CONCLUSIONRepeat sequence and strong topoisomerase II cleavage site around the breakpoint may predispose double-strand DNA breaks and recombination, which, in addition to the nonhomologous end-joining mechanism, may contribute as important factors to the gene deletion.

Adult ; Base Sequence ; Chromosome Breakage ; Cloning, Molecular ; Dystrophin ; genetics ; Exons ; genetics ; Gene Deletion ; Humans ; Male ; Molecular Sequence Data ; Muscular Dystrophies ; genetics ; pathology ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo evaluate the endourethral surgery for the complicated urethra stenosis and urethratresia.

METHODSThe endourethral surgery, such as internal urethrotomy transurethral scar electrosectomy or transurethral scar plasmakinetic bipolar electrocautery (PKR) or transurethral laser cicatrectomy, were carried out in 46 cases suffering from the complicated urethra stenosis and urethratresia.

RESULTSThe curative rate in this series being achieved by once and twice or three times'operation were 80.43% (39/46) and 13.04% (6/46) respectively. Three cases of treatment failure were caused by long-segment stricture and urethratresia or severe malposition of the urethral proximal and distal to a narrow-caliber area or post-operation infection. Thirty-nine cases have been followed up for 6 to 84 months. Satisfactory voiding has been achieved in all patients.

CONCLUSIONEndoscopic surgery was believed to be a safe and efficient therapeutic choice for the complicated urethra stenosis and urethratresia. The success of the treatment depends on understanding the length of the stricture before operation, resecting completely the scar tissue with electric or PKR or laser technique during the process, preventing infection and managing appropriately the urethral catheterization after operation.

Adolescent ; Adult ; Aged ; Endoscopy ; Follow-Up Studies ; Humans ; Laser Therapy ; Male ; Middle Aged ; Retrospective Studies ; Urethra ; abnormalities ; surgery ; Urethral Obstruction ; surgery ; Urethral Stricture ; surgery ; Urogenital Surgical Procedures ; methods

OBJECTIVETo study the mechanisms of dystrophin gene deletion, the junction fragment with exons 45-54 deletion were cloned and sequenced.

METHODSA Duchenne muscular dystrophy (DMD) patient with exons 45-54 deletion has been substantiated by PCR amplification of the exons. Then we used a PCR-based genome-walking method for localizing the breakpoints in introns 44 and 54. At last, the deletion-junction fragment was directly amplified by PCR approach with forward and reverse primers annealing to a DNA sequence as close as possible to the breakpoints in introns 45 and 54. The sequencing result of the deletion-junction fragment was compared with the normal intronic sequences.

RESULTSA total of 2716 bp sequence containing the junction fragment was obtained. The 5' breakpoint was located in LINE/L1 element of intron 44 and close to a matrix attachment region (MAR). The 3' breakpoint was located in the minor potential MAR with topoisomerase II cleavage sites around. Beside the 3' breakpoint there was a 6 bp palindromic sequence. A 4 bp microhomologous sequence (AGAG) was in the joint of the deletion-junction fragment.

CONCLUSIONThe nonhomologous recombination caused by L1 repeated element, topoisomerase II cleavage sites, MARs and the nonhomologous end joining of microhomologous sequence may be the important factors in this huge gene fragment deletion.

Base Sequence ; Child ; Cloning, Molecular ; DNA Topoisomerases, Type II ; metabolism ; Dystrophin ; genetics ; Exons ; genetics ; Humans ; Introns ; genetics ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Polymerase Chain Reaction ; Sequence Deletion