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.

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 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.

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.

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 synergetic effect of HBX protein and M2 macrophages in inflammatory microenvironment on invasion and metastasis of hepatocellular carcinoma cells.Methods Hep3B cells were infected with recombinant lentivirus carrying HBx gene,following co-culture with THP-1 original M2 macrophages.The cells were divided into six groups:two infected groups (Hep3B +and Hep3B + + M2),four non-infected groups (Hep3B-,Hep3B-+ LV5,Hep3B-+ M2,Hep3B-+LV5 + M2).Western blot (WB) was used to assess the expression changes of E-cadherin and N-cadherin,markers of epithelial-mesenchymal transition (EMT).The cellular location of EMT markers was observed by immunofluorescence confocal microscopy.Transwell assay was used to evaluate the invasion ability of Hep3B cells.Results HBX protein overexpressed in Hep3B cells by lentivirus infection.After 72 h co-culture with M2 macrophages,WB results showed that E-cadherin descreased significantly in Hep3B+ (0.42 ±0.11) when compared with Hep3B-(1.00 ±0.18) (t =4.762,P ＜0.05),while N-cadherin was significantly higher in Hep3B + (2.85 ± 0.44) than in Hep3B-(1.00 ± 0.17) (t =4.762,P ＜ 0.05).M2macrophages decreased E-cadherin expression in Hep3 B + + M2 (0.1 ± 0.13) compared with Hep3 B + (t =3.255,P ＜0.05),while N-cadherin expression increased in Hep3B+ + M2 (4.18 ± 0.52) (t=10.009,P ＜ 0.05).Non-Infected groups didn't change the markers of E-cadherin and N-cadherin.It was suggested that invasion ability of Hep3B increased by HBx overexpression.Conclusions HBX protein and M2 macrophages synergetically mediated the invasion and metastasis of hepatocellular carcinoma cells by EMT.

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.

This article was aimed to study anti-inflammatory effects of petroleum ether extract from Citrullus lanatus vine (PEECLV) and explore the mechanism. The methods of ear swelling with xylene, increasing of peritoneal capil-lary permeability, and paw edema with egg albumin were used in the establishment of mice models, respectively. Granuloma with cotton pellet, and paw edema with carrageenan were used in the establishment of rat models, re-spectively. Observation was made on anti-inflammatory effects of PEECLV. The content of nitrous oxide (NO) and nitric oxide synthase (NOS), as well as the level of TNF-α and IL-1β in blood serum were measured among granu-loma with cotton pellet rat models. Contents of malondialdehyde (MDA), prostaglandin E2 (PGE2), 5-HT, histamine (His), and protein in inflammatory exudates from the paw edema with carrageenan rat model, as well as contents of NO and NOS in blood serum, and activity of superoxide dismutase (SOD) were measured in order to study the possi-ble anti-inflammatory effects. The results showed that PEECLV can suppress the ear swelling with xylene, the in-creasing of peritoneal capillary permeability, and the paw edema with egg albumin in mice, as well as granuloma with cotton pellet, paw edema with carrageenan in rats. It had anti-inflammatory effects at different degrees. The content of NO and NOS, as well as the level of TNF-α and IL-1β in blood serum were reduced in granuloma with cotton pellet rat models. The content of MDA, PGE2, 5-HT, His, and protein content were reduced in inflammatory exudates from the paw edema with carrageenan rat model. The increasing of NO and NOS in blood serum was inhib-ited. And the activity of SOD was increased. It was concluded that PEECLV had certain anti-inflammatory effects. Its mechanism may be related to the influence of the production of inflammatory mediators and antioxidant.

Objective To explore the interleukin (IL)-6,IL-10 and T cell subsets levels in bronchoalveolar lavage fluid(BALF) of children with refractory mycoplasma pneumonia.Methods A total of 53 children with refractory mycoplasma pneumonia were selected as the observation group,30 children with bronchial foreign body in our hospital were chosen as controls during the same period.ABC-double antibody sandwich ELISA method was used to detect IL-6,IL-10 levels and the CD3 +,CD4 + and CD8 + T levels were measured by means of flow cytometry in BALF.Results The IL-6 and IL-10 levels in BALF of children in the observation group were (63.25 ± 18.61) ng/ml,(31.83 ± 8.33) ng/ml respectively,and they were significantly higher than those of the controls[(30.51 ± 1.34) ng/ml,(11.01 ± 2.91) ng/ml] (P ＜ 0.05,respectively).The percentage of CD3 +,CD4 +,CD8 + T cells and the ratio of CD4 +/CD8 + T cells in BALF of the observation group were (48.47 ± 2.88)％,(21.16 ± 6.29)％,(23.04 ± 4.63)％,0.94 ± 0.33,respectively,and they were significantly lower than those of the controls [(64.24 ± 3.06) ％,(34.34 ± 7.59) ％,(26.71 ±5.29)％,1.56-±0.67] (P＜0.05,respectively).Conclusion The IL-6,IL-10 levels in BALF of children with refractory mycoplasma pneumonia significantly increased,suggesting that cell-mediated immunity play an important role in the pathogenesis of refractory mycoplasma pneumonia.

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 .