Objective To study the characteristics of mutations of gene COL4A5 encoding type Ⅳ collagen ? 5 chain in one chinese pedigree with X-linked dominant inherited nephritis (Alport's syndrome,AS). Methods Genomic DNA was extracted from 35 members of the pedigree of Alport's syndrome. All of 51 exons of COL4A5 gene were amplified by PCR with the primers synthesized according to the published intron sequences of COL4A5. The PCR products were further analyzed by agarose gel electrophoresis and single strand conformation polymorphism (SSCP). The PCR products showing polymorphism were directly sequenced. Results PCR-SSCP analysis showed that 17 PCR products had abnormal mobility of single strand DNA. DNA sequencing analysis revealed 9 suspicious mutations. But these suspicious mutations were not be confirmed by inverse sequencing analysis. Conclusion The exon mutation of COL4A5 gene of this pedigree did not be found, and the mutations of COL4A5 gene may locate in the its introns.

Objective To explore the safety and clinical effects of laparosepie splenectomy.Methods Clinical data of 24 cages of laparoscopic splenectomy in our hospital from 2002 to 2008 were retrospectively analvzed.Among these 24 cases,there were 6 cases with liver cirrhosis,10 cases with ITP,2 cases with hemolytic anemia(Evens syndmm),2 cases with spelenic rupture,and 4 cases optimum spleen ambuty.Results All the 24 cases were successfully underwent laparoscopic splenectomy.The mean operation time wag 146 minute.the mean volume of blood loss was 220 ml.the postoperative gastrointestinal peristalsis time Was from 24 to 48 hours.The mean hospitalization time was 9 days after operation.Conclusions Provided mastering operation indication and technique,Laparoscopie spleneetomy is a safe and minimally invasive surgery.

BACKGROUND:Although the mechanism why neuronal cels wil die after transient cerebral ischemia has not been completely elucidated, many researches nowadays have investigated the pathological mechanism in the level of celular organs, such as mitochondria.
OBJECTIVE:To summarize and discuss the functions of neuronal mitochondria and apoptosis signaling pathways in transient cerebral ischemia.
METHODS: A computer-based online retrieval was performed to search papers in CNKI and PubMed databases using the key words of “cerebral ischemia, mitochondrion, apoptosis, reactive oxygen species, reperfusion, superoxide dismutase, nitric oxide synthase, Bcl-2 protein family, review” in Chinese and English, respectively. Papers published recently or in the prestigious journals were selected in the same field. After excluding objective-independent papers and repeated studies, 50 papers were included for further analysis.
RESULTS AND CONCLUSION:Recently mitochondria are found to play an important role after transient cerebral ischemia by producing a lot of reactive oxygen species to activate many kinds of signaling pathways and regulate mitochondria-mediated apoptosis. Reactive oxygen cannot only induce biomacromolecule injury but also induce apoptosis signal transduction. Deeply investigation is needed on the pathological mechanism after transient cerebral ischemia.

Objective Research has indicated that hydrogen sulfide(H2S) can regulate the function of N-methyl-D-aspartate re-ceptors(NMDARs) in the brain, but its effect on brain resuscitation requires further investigation.The study was to speculate the effect of H2 S on brain resuscitation as well as the underlying mechanism of neuroresuscitation by investigating the effects of hydrogen sulfide and hypo-thermia on the expression of NR2A, NR2B and phospho-cAMP response element binding protein (p-CREB) of NMDARs in the hippocampus after global cerebral ischemia following by reperfusion. Methods 100 male SD rats were randomly divided into five groups(n=20):sham operation group, model group, mild hypothermia group, NaHS group, NaHS combined mild hypothermia group.Pulsinelli-Brierley four-ves-sel occlusion method was induced to build the injury rat model by reperfusion after global cerebral ischemia .After 15 minutes'ischemia, im-mediate injection of 14μmol/kg NaHS was performed intraperitoneally on NaHS group and NaHS combined mild hypothermia group , while skin cooling(rectal temperature=32-33℃) was done on mild hypothermia group and NaHS combined mild hypothermia group .6 hours late,r hip-pocampus were extracted from rat heads.Respectively, spectrophotometer was applied to measure the content of H2S, Western blot for the expres-sions of NR2 A,NR2 B and pC-REB, and RTP-CR for mRNA level of brain derived neurotrophic (BDNF). HE staining was also performed on brain tissues 72hours after reperfusion on 4 rats from each group to evaluate the pathological changes of pyramidal neurons in CA1 region. R esul ts The content of H 2 S increased in each of the four groups after ischemia-reperfusion compared with sham operation group ( 15.2 ±2.0 nmol/g) (P<0.05).In comparison to model group (25.2 ±3.5 nmol/g), NaHS group (37.5 ±4.0 nmol/g) and NaHS combined mild hypothermia group (38.7 ±4.4nmol/g ) resulted in significant high content of H2S(P<0.05), while mild hypothermia group(26.5 ±3.5nmol/g ) got a mild increase(P>0.05).The gray values of NR2A and NR2B in each group increased compared with sham operation group(P<0.05), re-sulting in NR2A/NR2B<1 in model group and mild hypothermia group while NR2A/NR2B>1 in NaHS group and NaHS combined mild hy-pothermia group.Compared with the expression of p-CREB(0.55 ±0.06) in model group, there were significant increases in mild hypother-mia group(0.99 ±0.15), NaHS group(1.05 ±0.12), NaHS combined mild hypothermia group(1.02 ±0.15)(P<0.05).Compared with the expression of BNDF mRNA(0.83 ±0.12) in model group, there were significant increases in mild hypothermia group (1.11 ±0.13), NaHS group(1.27 ±0.16), NaHS combined mild hypothermia group(1.35 ±0.16)(P<0.05).In comparison to model group, there were signifi-cant alleviation in the injury of pyramidal neurons in hippocampal CA1 region in mild hypothermia group, NaHS group, NaHS combined mild hypothermia group, with the best effect in NaHS combined mild hypothermia group . Conclusion Hydrogen sulfide combined mild hypo-thermia can selectively activate synaptic NMDA receptors and trigger the prosurvival CREB signaling pathway to exert brain resuscitation .

Objective To compare the impacts of subtotal nephrectomy and ischemiareperfusion injury on renal stem cells and progenitor cells of rats,and to explore the significance of renal stem cells and progenitor cells for renal repair and the possible mechanisms of prognosis in rats with acute renal failure (ARF) or chronic renal failure (CRF).Methods Rats of CRF or ARF model underwent 5/6 nephrectomy or renal artery ligation and repedusion respectively,and rars in control group underwent sham operation.Scr,BUN and 24 hour urine protein were regularly measured.Kidney specimens were obtained at the set time for HE staining and fluorescence staining.Expressions of CD24,CD133 and podocin were detected by immunofluorescence.RT-PCR was performed to quantify the expression of transforming growth factor β1 (TGF-β1),Notch2,hepatocyte growth factor (HGF),bone morphogenetic protein 7 (BMP7) and Pax-2 mRNA in renal tissue and the expression of podocin mRNA in renal cortex.Correlation among the expressions of Pax-2 mRNA,podocin mRNA and glomemlosclerosis index were analyzed.Results The rats of two models presented typical ARF or CRF in renal pathology and function.Glomerulosclerosis index in CRF group increased gradually with time,which were (2.34±0.28)％,(25.12±5.67)％,(89.42±12.28)％ and (171.23±32.28)％ at day 14,day 30,day 60 and day 90 respectively.Compared with sham group,the CD24+CD133+ cells of the ARF rats showed no significant change in quantity and distribution,while the CRF rats showed gradual reduction of CD24 +CD133+ cells.The expression of podocin in glomerulus decreased temporarily and recovered finally after ischemiareperfusion injury,but decreased gradually after 5/6 nephrectomy.Compared with sham group,expression of TGF-β1,Notch2 mRNA in renal tissue was increased in CRF group,while the expression of HGF,BMP7 mRNA in renal tissue of ARF group were increased.Between the expression of Pax-2 mRNA in renal tissue and the expression of podocin mRNA in renal cortex,there was positive correlation in CRF group,while they both were negatively correlated with glomerulosclerosis index.Conclusions Ischemia-reperfusion injury makes no obvious impairment to renal progenitor cells.Having progressively injured the living environment of renal progenitor cells,subtotal nephrectomy reduces renal progenitor cells,and causes podocytes to repairing incompetently,which may be the main pathogenesis of CRF with poor prognosis.

Objective To evaluate the accuracy of thoracic pedicle screw placement using the "funnel technique" and investigate its consistency in experimental study and clinical application.Methods The clinical data of three human cadavers and 11 patients with the installation of at least one thoracic pedicle screw at T1 -T12 from August 2006 to July 2008 were retrospectively analyzed. One junior spine surgeon lack of experience were responsible for placing these screws with the "funnel technique".The accuracy of screw placement and the complications related to the use of thoracic pedicle screws were analyzed by assessing postoperative CT scans. Results The mean follow-up time was 23.1 months,which showed no vascular or visceral complications, or iatrogenic neurological injury. The rate of unintended cortex perforations was 14% (10/72) in cadavers and 15% (8/55) in patients, respectively.The critical perforation occurred in two screws (3%) in cadavers and one screw (2%) in patients. There was no statistical difference between the percentage of cortex perforations in cadavers and patients. Of all the 11 patients, screw violation occurred laterally in six ( 11% ), medially in one ( 2% ) and superiorly in one (2%). No violations occurred inferiorly or anteriorly. For all patients, only one screw needed revision. The perforations made by the junior spine surgeon occurred in six screws in the first cadaver, three in the second cadaver and one in the third cadaver. Conclusions The "funnel technique" is a simple,safe, accurate and cost-effective technique for pedicle screw placement. The result of the experimental study is consistent with that of the clinical application. "funnel technique" is helpful for junior spine surgeons to master the technique of thoracic pedicle screw placement.

OBJECTIVETo understand the embryo after-ripening phenomenon of Panax quinquefolium, the growth and development process of fruit and seed was investigated in this study.

METHODThe growth and development characteristics of fruit and seed were obtained by field observation, the morphological changes were measured with a vernier caliper, paraffin section was used as well.

RESULT AND CONCLUSIONThe plant reached the most flowering numbers in 5-8 d after initial blooming time and the lag phase of embryo occurred in about 70 d. The size, fresh and dry weight of fruit and seed were all reached maximum in fruit maturation period. As the result showed, the development of seed was stopped at torpedo form of embryos, this conclusion can be applied to explore the morphological after-ripening mechanism of P. quinquefolium seed.

In this study, the rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry ( RRLC-QTOF/MS ) was used to profile the metabolites of urine samples from Childhood Pneumonia ( CP) patients and healthy controls and find the potential biomarkers which can support evidence to early diagnose and cure the disease. Choose 10 CP patients ( age 47. 72 ± 2. 35 months) and 10 healthy controls ( age 46 . 65 ± 1 . 97 months ) . The urine samples were analyzed by RRLC-QTOF/MS and then the resulting data matrices were analyzed by principal components analysis ( PCA ) to find the potential biomarkers. Urine samples of CP patients were successfully distinguished from those of healthy controls. A total of two significantly changed metabolites have been found and identified as potential biomarkers. It is suggested that the disorder of purine metabolism and amino acid metabolism may play an important role in the mechanism of CP.

Objective To summarize the clinical characteristics of neonates with non-immune hydrops fetalis.Methods The clinical data of ten neonates with non-immune hydrops fetalis, who were admitted to the Department of Neonatology, Gynecology and Obstetrics Hospital of Fudan University between January 2012 and June 2014, were retrospectively studied.Prenatal characteristics, causes, clinical features and prognosis were explored and analyzed with descriptive statistical methods.Results (1) One of the ten neonates was diagnosed after birth presented with hydroderma and abdominal distension, and the rest ones were diagnosed antenatally.There were six males and four females, and eight premature and two term neonates with the average gestational age of (33.6±2.4) weeks and birth weight of(2 680±478) g.(2) The mean maternal age was (30.3 ±4.6) years.Two of the mothers had gestational diabetes mellitus and one had gestational diabetes mellitus complicating with hyperthyroidism.The blood types of all mothers were O and Rh(+).None of the mothers were tested for parvovirus B19 and no fetus was reported with abnormal chromosome karyotype in the five cases received fetal karyotype analysis.(3) The average gestational age at initial identification of hydrops fetalis by fetal ultrasound was (31.3 ± 2.4) weeks (25.1～37.0 weeks) among nine cases diagnosed prenatally.By ultrasound screening, there were seven cases with hydrothorax, six with seroperitoneum, five with polyhydramnios and four with hydroderm.(4) Two neonates had normal non-stress test results during labor, and eight were abnormal and suffered from severe neonatal asphyxia at birth and resuscitated by endotracheal intubation.Ten neonates had hydroderm, seven had hydrothorax, six had seroperitoneum and one had hydropericardium.One complicated with multiple malformations and one had chromosome abnormality.Four cases received thoracentesis and three had abdominal paracentesis after parturition.(5) Prognosis: One neonate survived and nine died among which six due to resuscitation failure in delivery room, two died one day after giving up treatment after one day, and one died due to treatment failure six months after birth.(6) The causes of hydrops fetalis were anemia (two cases) and congenital diaphragmatic hemangioma, recurrent atrial premature beat, Down's syndrome, congenital pulmonary lymphangiectasia, paroxysmal supraventricular tachycardia, placental chorioangioma, suspicious genetic syndrome and idiopathic (one for each).Conclusions The prognosis varies because of different etiology of non-immune hydrops fetalis.Neonates with hydrops fetalis might have a higher rate of asphyxia and mortality rate, and difficulties in resuscitation at birth.

Objective To explore the mechanism of the role of mitomycin C(MMC)in regulating miR-200b expression and inducing fibroblasts apoptosis. Methods Fibroblasts cultured in vitro were treated with different concentrations of MMC for 5 min and continue culture for 24 h. The expression of miR-200b were analyzed by Real-time PCR. Cell apoptosis were observed using TUNEL staining. The expression of cleaved-PARP,Bax and Bcl-2 were detected by Western blot. The methylation level of miR-200b promoter were measured by BSP. Results After treated with MMC,The expression of miR-200b significantly downregulated.TUNEL Staining analysis demonstrated MMC could significantly induce human fibroblasts apoptosis. Western blot results showed cleaved-PARP,Bax increased and Bcl-2 decreased.The methylation ratio of miR-200b promotor increased and has a significant dose dependent. Conclusion MMC induced human fibroblasts apoptosis by promoting miR-200b promoter methylation.