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.

Objective:To explore the use of internet-based continuous visual recognition task (MemTrax test, MTX) as a rapid screening tool for amnestic mild cognitive impairment (aMCI).Methods:Sixty-four patients with aMCI and 64 individuals with normal cognition as healthy controls were enrolled respectively from Department of Neurology and Health Examination Center of the First Affiliated Hospital of Kunming Medical University from August 2018 to December 2019. Montreal Cognitive Assessment (MoCA) scale and MTX were adopted to assess the cognitive function of all subjects. The total adjusted MoCA scale score, correct rate of MTX, reaction time of MTX and MTX score were obtained and statistically analyzed.Results:The adjusted MoCA scale scores of aMCI patients and healthy controls were 19 (14, 24) and 26 (24, 27; Z=6.795), the correct rate of MTX of aMCI patients and healthy controls were 74% (60%, 80%) and 88% (84%, 94%; Z=8.359), and the MTX score of aMCI patients and healthy controls were 51.11±14.07 and 70.56±14.91 ( t=7.590), respectively, all with statistically significant difference ( P<0.001). Reaction time of MTX of aMCI patients and healthy controls was 1.401 (1.253, 1.590) s and 1.277 (1.163, 1.410) s, respectively ( Z=3.083, P<0.01). After adjustment for age, physical or mental occupation, exercise, hypertension, hyperlipidemia, stroke, sleep time, as well as smoke, the linear regression showed that the aMCI patients had a significant decrease of adjusted MoCA score, correct rate of MTX and MTX score ( P<0.001), and an extension of reaction time of MTX ( P=0.071), compared with the controls. By MTX and MoCA scale assessment, the best cutoff value was 81% for correct rate of MTX and 23 for adjusted MoCA scale score respectively for the prediction of aMCI (with sensitivity of 79.7%, 93.8% respectively, and specificity of 68.8%, 82.8% respectively). The area under the curve (AUC) of correct rate of MTX was 0.93 (95% CI 0.89-0.97, P<0.001), and the AUC of adjusted MoCA score was 0.85 (95% CI 0.78-0.91, P<0.001). There was a statistically significant difference in paired comparison of the two AUCs (χ2=4.620, P<0.05). Conclusion:MTX acts better for the detection of aMCI than MoCA scale, and correct rate of MTX<81% can be considered as the existence of MCI.

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.

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 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.

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 understand the occurrence and the related risk factors of birth defects. Methods Descriptive analysis was conducted on birth detect surveillance in the infants during January 2008 to June 2014. Results A total of 777 cases of birth defect were detected in 73498 infants, and the incidence of birth defect was 1.06%. The 5 most common birth de-fects were congenital heart disease, multi ifnger (toe), hypospadias, cleft lip, and palate and deformity of external ear. Compared infants born with no birth defects, male, preterm, low birth weight, twin and multiple births and resident were statistically higher in infants with birth defects (P<0.05). The major risk factors of birth defects were the medication history, spontaneous abortion, gestational diabetes mellitus, and family history. Conclusions The incidence of birth defect can be reduced by providing good health care during pre-marriage and pregnant so as to decrease the occurrence of premature infants, twins and multiple births, and low birth weight as well as improving prenatal diagnosis and intensifying birth defects surveillance.

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.