Background Rhodopsin (RHO) gene is the most common disease gene for autosomal dominant retinitis pigmentosa (adRP),one of the main pathogenesis is that misfolded mutant RHO proteins accumulate in the endoplasmic reticulum and cause endoplasmic reticulum stress (ERS).Objective This study aimed to determine the genetic basis for a consanguineous Chinese Han adRP family.Methods This study procedure complied with Helsinki Declaration.All participants in the family were investigated under the informed consent.Regular ocular examination was performed on the patients in this family.Next-generation sequencing (NGS) was carried out to screen the mutations in 189 genes associated with hereditary retinal diseases (HRDs).After being analyzed and filtered,variations detected by NGS were validated by Sanger sequencing and evaluating of pathogenicity.The wild-type RHOWT and mutant RHOP53Rwere cloned into the vector pEGFP-N1.Then the two plasmids were transfected into adult retinal pigmentosa epithelium cell line(ARPE19) and human embryo kidney 293 line (HEK293) to observe the location of rhodopsin-GFP fusion protein in cells,and the expression of ERS related protein XBP1 in the cells was detected by quantitative-PCR and Western blot.Results This family included 5 generations with the typical adRP characteristics.Genetic analysis identified a heterozygous variation,p.P53R in RHO gene,which was fully cosegregated in the family.Wild-type RHOWT-GFP fusion proteins showed the green fluorescence on the endoplasmic reticulum and cytomembrane,but the misfolded mutant RHO-GFP fusion protein gathered only in endoplasmic reticulum.Compared to wild-type RHOWT,the XBP1 was activated and increased by (1.28 ±0.09) fold.The introns of 26 bases in XBP1 mRNA were removed in the HEK293 cells with mutant RHO-GFP fusion protein,and the expression of XBP1 was stronger in the HEK293 cells with mutant RHO-GFP than that in HEK293 cells with wild type RHO-GFP and cells with blank pEGFP-N1 plasmid.Conclusions Heterozygous variant RHO p.P53R is very likely the pathogenical mutation in the adRP family.The RHOP53R mutant rhodopsin protein can not be delivery effectively from the endoplasmic reticulum to the cell membrane,and these proteins accumulate in the endoplasmic reticulum,which causes ERS.

OBJECTIVETo study the effect of stigma maydis polysaccharide (SMPS) on gastrointestinal movement.

METHODTaking charcoal as the indicator and taking ratio of charcoal movement, beginning time of black excretion and stool amount as the index to observe the effect of SMPS on intestinal movement in mice. Taking emthylorange as the indicator and taking the ratio of residual rate of methylorange as the index to observe the effect of SMPS on gastric emptying in mice. Taking methylene blue as the indicator and taking the time of gastric emptying and movement speed of intestinal content as the index to observe the effect of SMPS on gastrointestinal movement in rats. Observing the changes of cholecystokinin (CCK) level in plasm in rats.

RESULTCompared with control, the ratio of charcoal movement increased in mice (P <0.01). The beginning time of black excretion shortened and the stool amount increased in mice (P <0.01). The ratio of residual rate of methylorange increased in mice (P <0. 01). The time of gastric emptying prolonged in rats (P <0.01). The movement speed of intestinal content in rats accelerated (P <0.01). CCK level in plasm increased in rats (P < 0.05).

CONCLUSIONEffects of stigma maydis polysaccharide on gastrointestinal movement are probably related to the increasing of CCK level in plasm.

Animals ; Cholecystokinin ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Gastric Emptying ; drug effects ; Gastrointestinal Agents ; isolation & purification ; pharmacology ; Gastrointestinal Motility ; drug effects ; Intestine, Small ; physiology ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Zea mays ; chemistry

Objective To evaluate the clinical effect and safety of percutaneous closure of patent foramen ovalein treatment of migraine.Methods We studied 23 patients with migraine who were tested positive in Contrast Transcranial Doppler (TCDc) at the Department of Neurology in Bethune First Hospital of Jilin University between 2010 May and 2011 November.Procedures of transcatheterclosure of patent foramen ovale were performed at Department of Cardiovascular Surgery.At varied time points post-surgery,the patients were re-examined by TCDc and transthoracic echocardiograph (TTE) and followed up by phone interviewed with Headache Impact Test-6 (HIT-6) questionnaire to elevate improvement of migraine.The data was analyzed statistically.Results Percutaneous closure was successful in all patients without major long-term side-effects.At follow-ups at 48-542 days ((247 ± 145) days) post-surgery,the scores of HIT-6 were reduced to 38-70 (51 ± 9),compared to the pre-operative scores of 42-76 (61 ± 8).Disability by migraine had significantly improved (t =4.80,P ＜ 0.05).In 6 patients with migraine with aura,the scores of HIT-6 were reduced to 46-56 (51 ± 3),from preoperative scores of 63-76 (69 ± 4) and their disability affected by migraine had significantly improved (t =6.80,P ＜0.05).In 17 patients with migraine without aura,the scores of HIT-6 after surgery were reduced to 36-70 (52-10) from the preoperative scores of 42-69 (60 ± 8) and disability affected by migraine in this group had significantly improved (t =3.77,P ＜0.05).Among 18 patients having MRI or CT examination before the operation,8 patients had neurological events (such as cerebral infarction,cerebral ischemia) with average age of (44 ± 7) years old.In this group,disability affected by migraine had significantly improved (the HIT-6 scores:49 ± 7 vs 58 ± 10,t =2.49,P ＜ 0.05).Conclusion Percutaneous closure of PFO showed significant improvement in migraine treatment and it is a safe procedure.

Objective To explore the clinical efficacy of early surgery combined with electron linac therapy on keloid.Methods The keloid patients with stable phase were selected;complete resection of keloid and relaxation suture were performed;after the surgery within 24 hours 6 MeV with Varian 2300CD radiotherapy was given,each measuring 4 Gy,total dose of 20 Gy.Results 860 cases of patients were colected for a period of 3 months to 36 months of regular follow-up,which recovered in 802 cases (cure rate was 93.26％),effective results were observed in 41 cases (effective rate of 4.77％),including 17 cases of recurrence (recurrence rate 1.98％),the total efficiency (cure plus effective) was 98.02％.Conclusions More accurately immediate radiotherapy after surgery can effectively reduce the recurrence rate,which is a safe and effective method in the treatment of keloids.

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

OBJECTIVEWernicke's encephalopathy (WE) is an acute neuropsychiatric syndrome resulting from thiamine deficiency, which is associated with significant morbidity and mortality. The disorder is still greatly underdiagnosed in children because of either a relatively non-specific clinical presentation in some cases or unrecognized clinical setting. The aim of this literature review was to provide knowledge of pediatric WE in an effort to assist in early diagnosis, thereby reducing the morbidity and mortality.

METHODSThe clinical manifestations, characteristic magnetic resonance imaging (MRI), diagnosis and treatment of one case and the other 35 cases reported in the last decade in children were summarized.

RESULTSThirty-six cases (22 boys and 14 girls, 2-month to 16-year-old) were analyzed. All the other 35 cases except for our case had underlying diseases: improper feeding in 25/35 cases, long-time vomiting in 5/35 cases, immunosuppressive therapy in 4/35 cases, long-time total parenteral nutrition without multivitamin preparations supplementation in 3/35 cases and anorexia nervosa in 1/35 case. The classic triad (mental-status changes, nystagmus and ophthalmoplegia, and ataxia) was seen in 6/36 cases. The other clinical manifestations included consciousness disturbance in 24/36 cases, infection in 22/36 cases, pathological reflex and muscular tension changes in 18/36 cases, convulsion in 17/36 cases, developmental delay in 4/36 cases and failure to thrive in 2/36 cases. Cerebrospinal fluid examination was performed in 31/36 cases, and a slightly raised protein concentration was seen in 7/31 cases. The cerebrospinal fluid lactate levels were detected in 4/36 cases (all increased), serum lactic acid levels in 7/36 cases (6/7 cases increased), serum pyruvate in 4/36 cases (all increased), thiamine pyrophosphate effect (TPPE) in 9/36 cases (all increased), and serum thiamine in 2/36 cases (increased in 1/2 cases). The brain computed tomography (CT) scan was conducted in 20/36 cases and 16/20 cases showed abnormal hypodensity in bilateral basal ganglia, one case revealed diffuse cortical atrophy. The brain MR scan was conducted in 13/36 cases and all the 13 cases revealed symmetrical abnormal signal in bilateral mamillary body and basal ganglia, and 7/13 cases showed abnormal signals in the tegmentum of midbrain, cerebral aqueduct and white matter around the third and fourth ventricles. The diagnosis of WE was confirmed by MR in 12 cases, triad combined with MR in 3 cases, autopsy in 1 case among the 13 cases who underwent MR scan. The diagnosis of WE was confirmed by the TPPE and/or lactate levels in 9/11 cases. The initial thiamine was given by intravenous or intramuscular infusion in 33/36 cases, unknown method in 1 case, orally in 1 case and no thiamine was used in 1 case. The dosage of thiamine was 100 mg daily in 29/35 cases, unknown in 3/35 cases, 50 mg daily in 2/35 cases, 600 mg daily in 1/35 case. 34/35 patients' clinical symptoms improved during 24 hours to 1 week after initial treatment, and 1 case died due to no response to thiamine. Nineteen patients were followed up for 2-2.5 months and 17 cases recovered completely.

CONCLUSIONWernicke's encephalopathy can be difficult to diagnose because of a relatively non-specific clinical presentation. The characteristic MRI findings and the dramatic response of neurological signs to parenteral thiamine will assist early clinical diagnosis. Early and timely thiamine supplementation could reverse the clinical features and improve the prognosis in most cases.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Sepsis ; complications ; Wernicke Encephalopathy ; complications ; diagnosis

To investigate changes of 14-3-3beta from apoptosis induced by PrP106-126 polypeptide, HeLa cell was incubated with PrP106-126 for 4h or 8h. Nucleus changes and the expression of PARP were detected differently by Hoechst staining and Western blotting. Expressing of protein and mRNA from 14-3-3beta was determined by Western blotting and Real-time PCR. The results show that typical nucleus pyknosis and chip of apoptosis and degradation of PARP were induced by PrP106-126 peptide in HeLa cells. Degradation of 14-3-3beta appeared in apoptosis groups induced by PrP106-126 peptide. However, 14-3-3beta mRNA did not display any changes in apoptosis groups. This study indicated that degradation of antiapoptosis protein 143-3beta induced by PrP106-126 peptide may be one of pathogenesis mechanism of prion disease.
14-3-3 Proteins ; metabolism ; Apoptosis ; drug effects ; HeLa Cells ; Humans ; Peptide Fragments ; pharmacology ; Prions ; pharmacology ; Proteolysis ; drug effects

Objective To observe the injury of high intensity focused ultrasound combined with radiotherapy (RT) to the pancreas,the unintended abdominal tissues and organs of swine in vivo.MethodsAccording to the criterion to grade the gross and histological injury of the pancreas and the unintended tissue,the scores of injury between the groups to get the data of safety and feasibility of high intensity focused ultrasound(HIFU)combined with RT were compared.Results There was a better tolerance in each group and no fatal complication was observed.For the pancreas there was significant difference for the combined group compared with the other groups.For the unintended target tissue there was no significance difference except the control group with the other groups.Conclusion HIFU combined with RT can increase the injury to the pancreas of the swine compared with HIFU alone; while there was no increase for the injury to the unintended target tissue.The main toxicity of combination treatment is the toxicity of RT and can be tolerated by the animals.If the HIFU treatment is strictly controlled,the combination of HIFU and RT is safe and feasible.

Background Statistic data revealed that different retinopathy of pre-term infants have different susceptibility to retinopathy of prematurity (ROP),which may be associated with polymorphism of vascular endothelial growth factor(VEGF) gene.Objective This study was to determine the association of polymorphisms of VEGF gene with the risk for ROP.Methods This research was approved by Ethics Committee of Hunan Children's Hospital,and written informed consent was obtained from the parents of patients.A prospective case-controlled study was designed.Ninety-nine ROP patients in Hunan Children' s Hospital and 88 pre-termed children without ROP were included from January,2006 to December,2009.Thirty-nine patients who received retinal photocoagulation or cryotherapy were included as the treatment group,and 60 untreated but spontaneously regressed ROP patients as the non-treatment group.No significant differences were seen in demography between with the ROP group and the without ROP group,or between the treatment group and the non-treatment group (all P＞0.05).2 mL of peripheral blood was collected for the extraction of DNA.Gene polymorphisms of VEGF-A+405 and VEGF-A936 were detected using the pyrosequencing method.Results No significant difference was found in the frequencies of the VEGF-A+405 gene polymorphisms between the ROP group and without ROP group (P =0.071,OR =0.675,95 ％ CI =0.444-1.026).Also no significant difference was found in the frequencies of the VEGF-A936 gene polymorphisms between with the ROP group and without the ROP group (P =0.118,OR =0.768,95 ％ CI=2.823-4.614).However,the frequencies of the VEGF-A+405 gene polymorphisms were significantly higher in the ROP treatment group than the non-treatment group (P＜0.01,OR--0.857,95 ％ CI =5.239-14.024),and VEGF-A936 gene polymorphisms was also significantly higher in the ROP treatment group than the non-treatment group (P =0.000,OR =3.609,95 ％ CI =0.711-0.829).Conclusions There is no association between the VEGF-A+405/VEGF-A936 single nucleotide polymorphism with the risk of ROP,but polymorphisms of VEGF gene may be correlated with the prognosis of ROP.The carrier of VEGF-A +405 /VEGF-A936 allele is more susceptible to ROP progression.

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.
Animals ; Egg Proteins ; genetics ; metabolism ; Electroporation ; Fermentation ; Immunization ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Rabbits ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; secretion ; Swine ; Zona Pellucida Glycoproteins