As one of the most serious hereditary neuromuscular disease, spinal muscular atrophy (SMA) is caused by the loss or mutation of survival motor neuron 1 (SMN1) gene. It leads to a decrease in the level of SMN protein and a consequent loss of alpha neurons and progressive muscle atrophy resulting in the progressive muscle weakness, the significant disability and the shortened lifespan. Up till now, only three drugs have been approved for SMA, including the gene therapy drug onasemnogene abeparvovec. The antisense oligonucleotide drug nusinersen and and the small molecule chemical drug risdiplam were briefly introduced. Some representative samples of the small molecule chemical drugs and antisense oligonucleotide drugs targeting SMN2 in the clinical trial or preclinical research phases were also reviewed.

Objective To investigate the influence of Helicobacter pylori(Hp) infection on gastric mucin MUC5AC and MUC6 expression in children.Methods Sixty-six cases with gastric biopsy specimens were obtained from 66 children undergone gastroscopy from Jan.2005 to Jun.2006 for episodic or continuous abdominal pain,nausea,vomiting,abdominal distension,retching and dyspepsia,and so on.Among these children,39 cases were male and 27 cases were female,owning a average age(8.8?3.0)years.These specimens were divided into 2 groups followed by the presence of Hp,which was detected by rapid urease tests and Hp-PCR.Gastric mucin MUC5AC and MUC6 mRNA were also measured by reverse transcription PCR(RT-PCR),and Hematoxylin and Eosin staining were used for pathology observation.Comparisons between every groups were performed using t test and ?2 test,and statistical significance was defined as P

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.

Objective To explore whether human umbilical venous endothelial cells could be used as feeder layer to support the growth of embryonic stem cells (ESC) and keep ESC undifferentiated.Methods The venous vessels of umbilical cord obtained from healthy puerperal were perfused with collagenase.The isolated endothelial cells went through primary culture and passages for expansion.Factor Ⅷ antigens determination was implemented.Endothelial cells with good growth and 3 or above passages were treated with mitomycin-C(10 mg/L) and prepared as feeder layer,on which E14.1 ESC was transplanted for subculturing to observe the morphological characterization and determine ESC alkaline psphatase (AKP) activity and the expression of stem cell marker Oct-4.Severe combined immune deficiency(SCID) mouse in vivo terotoma formation experiment was performed to identify its pluripotent properties.Results Human umbilical vein-derived endothelial cells grew well in culturing in vitro and regenerate in large numbers.The endothelial cells maintained normal cellular morphological and biological characterization after 10 passages.The cells stopped proliferating after being treated with mitomycin-C,but its activity and morphological properties were well-maintained with 24 hours,which was a fundamental property of serving as feeder layer.E14.1 ESC remained undifferentiated in human umbilical venous endothelial cells after 3-8 passages,the cells grew in colony and showed high expression of AKP and stem cell Oct-4.In vivo pluripotency experiment showed that 6 weeks after being transplanted to SCID mice E14.1 ESC of 6 and 10 passages in endothelial cells both could form teratoma containing 3 layers of tissue cells.Conclusions Human umbilical venous endothelial cell serve as a convenient feeder layer cell with rich sources.It can effectively support ESC growth and heterogenous and prevent the heterogeneous protein pollution and pathogenic microorganisms caused by animal cell feeder layers,thus solve the problem of biological safety of ESC clinical application.

Objective To determine the efficacy of hysteroscopy and laparoscopy in differential diagnosis of pregnancy-related diseases,including gestational trophoblastic neoplasia(GTN),incomplete abortion and ectopic pregnancy.Methods Twenty-seven patients with a suspected diagnosis of GTN were transferred to Peking Union Medical College Hospital from September 2003 to March 2006,and underwent hysteroscopy and laparoseopy.Clinical data of patients were reviewed retrospectively.Most patients had abnormal vaginal bleeding and persistently elevated plasma beta human chorionic gonadotropin(?-hCG) level for a median(53?37)days(range,15-125 days)after evacuation.Ultrasound revealed a lesion with affluent blood flow in intrauterine,unilateral horn of uterus,or myometrium.No positive findings were revealed by computerized tomography or X-ray of the chest in all patients.Eleven patients underwent evacuation under hysteroscope,10 patients were diagnosed and treated by laparoscopy,and 6 by hysteruscopy and laparoseopy.Results Choriocarcinoma was diagnosed in 4 patients,who achieved complete remission by chemotherapy later.The diagnosis of GTN was ruled out in the other 23 patients, including cornual pregnancy in 12,pregnancy in rudimentary horn in 1,and incomplete abortion in 10,who were cured by hysteroscopic and laparoscopic surgery and postoperative adjuvant single dose methotrexate.Conclusions The major causes of pregnancy-related abnormal bleeding include incomplete abortion,eetopic pregnancy,and GTN.Hysteroscopy and laparoseopy are effective alternative of diagnosis for differentiation of GTN from non-GTN and can also offer therapeutic treatment.

Objective To explore hemodynamic parameter changes in pulmonary arterial hypertension(PAH)treated by transplantation of bone marrow mesenchymal stem cells(MMSCs)in the experimental rats.Methods MMSCs cells were collected from bone marrow of Sprague-Dawleye(SD)rat's femoral and tibial bones,cultured and passaged in vitro,then stained by Hoechst 33342 fluorescence dye stuff.Ninety healthy male SD rats were randomly divided into 3 groups(n=30):normal control group(group N),MMSCs transplanted group(group M),PAH model group(group H).The rats in the two latter groups were given a single subcutaneous crotaline(50 mg/kg)to establish the model of PAH.The rats of group N were injected respectively a single subcutaneous 9 g/L saline water(6 mL/kg).After 21 days,5?109 L-1 MMSCs cultured in 1 mL phosphate-buffered saline were infused into the rats respectively in group M by sublingual vein and 1 mL L-DMEM was given in group H.The indexalso of right ventricle systolic pressure(RVSP),right ventricle hypertrophy index,arterial blood gas analysis and the changes of small pulmonary blood vessel were observed after 28 days.Results The administration of MMSCs 28 days after PAH nearly completely prevented the increase in RVSP with PAH alone [(32.20?2.32)mmHg vs(48.30?1.56)mmHg P

Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

Catalytic wet air oxidation (CWAO) of o-chlorophenol in wastewater was studied in a stainless steel autoclave using four different Fe catalysts in the temperature range of 100-200 degrees C. Experimental results showed that high rate of o-chlorophenol and COD(Cr) (Chemical Oxygen Demand, mg/L) removal by CWAO was obtained at relatively low temperature and pressure. The catalysts Fe2(SO4)3, FeSO4, Fe2O3 and FeCl3 all exhibited high catalytic activity. More than 93.7% of the initial COD(Cr) and nearly 100% of o-chlorophenol were removed at 150 degrees C after 150 min with FeSO4 as catalyst. The CWAO of o-chlorophenol was found to be pseudo-first order reaction with respect to o-chlorophenol, with activation energy of 75.56 kJ/mol in the temperature range of 100-175 degrees C.
Catalysis ; Chlorophenols ; analysis ; chemistry ; isolation & purification ; Industrial Waste ; prevention & control ; Iron ; chemistry ; Kinetics ; Models, Chemical ; Oxidation-Reduction ; Salts ; chemistry ; Solutions ; Temperature ; Water Purification ; methods

Acinetobacter baumannii is an important opportunistic pathogen in hospital, and the multidrug-resistant isolates of A. baumannii have been increasingly reported in recent years. A num-ber of different mechanisms of resistance have been reported, some of which are associated with plasmid-mediated acquisition of genes. Therefore, studies on plasmids in A. baumannii have been a hot issue lately. We have performed complete genome sequencing of A. baumannii MDR-TJ, which is a multidrug-resistant isolate. Finalizing the remaining large scaffold of the previous assem-bly, we found a new plasmid pABTJ2, which carries many phage-like elements. The plasmid pAB-TJ2 is a circular double-stranded DNA molecule, which is 110,967 bp in length. We annotated 125 CDSs from pABTJ2 using IMG ER and ZCURVE_V, accounting for 88.28%of the whole plasmid sequence. Many phage-like elements and a tRNA-coding gene were detected in pABTJ2, which is rarely reported among A. baumannii. The tRNA gene is specific for asparagine codon GTT, which may be a small chromosomal sequence picked up through incorrect excision during plasmid forma-tion. The phage-like elements may have been acquired during the integration process, as the GC content of the region carrying phage-like elements was higher than that of the adjacent regions. The finding of phage-like elements and tRNA-coding gene in pABTJ2 may provide a novel insight into the study of A. baumannii pan-plasmidome.

Great attention should be paid now to simultaneously removing common pollutants, especially inorganic pollutants such as nitrate and heavy metals, as individual removal has been investigated extensively. Removing common pollutants simultaneously by iron metal is a very effective alternative method. Near neutral pH, heavy metals, such as copper and nickel, can be removed rapidly by iron metal, while nitrate removal very much slower than that of copper and nickel, and copper can accelerate nitrate removal when both are removed simultaneously. Even a little amount of copper can enhance nitrate removal efficiently.Different mechanisms of these contaminants removal by iron metal were also discussed.