Brain Neoplasms ; pathology ; physiopathology ; DNA Repair ; genetics ; Glioma ; pathology ; physiopathology ; Humans ; Statistics as Topic ; Targeted Gene Repair

BACKGROUNDProteins or peptides can be directly transferred into cells when covalently linked to protein transduction domains (PTDs). TAT is one of the most widely studied PTDs. The effect of fusion protein TAT and heme oxygenase-1 (HO-1) on liver sinusoidal endothelial cells (SECs) apoptosis during cold storage is unknown. The present study aimed to determine whether fusion protein TAT-HO-1 would transduce efficiently into liver during cold storage, and, if so, to determine whether TAT-HO-1 would attenuate SECs apoptosis during preservation injury in rat.

METHODSLivers of Sprague-Dawley rats were harvested and randomly assigned to group 1 (HTK solution) and group 2 (HTK solution containing TAT-HO-1 fusion protein) according to the type of the preservation solution. The transduction efficiency of TAT-HO-1 was examined and the impairment of SECs was assessed during the period of cold storage followed by 1 hour of reperfusion.

RESULTSTAT-HO-1 can transduce efficiently into liver during cold storage. A significantly lower apoptotic index of SECs was observed in group 2, at 6, 12 and 18 hours of cold storage after 1 hour reperfusion, when compared with group 1. TAT-HO-1 reduced HA and ET levels in liver at each time point. Both Bcl-2 and Bax protein were expressed in hepatocytes and SECs at the periphery of the sinusoidal space. Moreover, higher Bcl-2 expression and lower Bax expression were observed in group 2.

CONCLUSIONSTAT-HO-1 can transduce efficiently into rat livers and shows a protective effect on SECs by attenuating apoptosis during cold ischemia/reperfusion injury. Protein transduction will be a novel therapeutic strategy to reduce the risk of preservation injury in liver transplantation.

Animals ; Apoptosis ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Heme Oxygenase-1 ; genetics ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Liver ; cytology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; tat Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVE: To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value. METHODS: Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control. RESULTS: Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method. CONCLUSION The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.
DNA/isolation & purification* ; DNA Fingerprinting ; Female ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Silicon ; Spermatozoa

OBJECTIVETo report two cases of glandular odontogenic cyst and examine its cytokeratin 18,19 expression.

METHODSTwo cases of glandular odontogenic cyst were reported and studied. The cytokeratin 18, 19 expression in these two cases were also investigated using immuno-histochemical staining as well as in the situ hybridization of the cyst epithelium.

RESULTSHisto-pathological examination revealed that ciliated columnar cells, squamous cells and low-columnar cells were found in the superficial layer of the lining epithelium. Several minor salivary glands, mainly composed of seromucous cells were observed near the satellite cyst. CK18 were expressed in all layers of the lining epithelium of varying intensity. CK18 was negative in lining epithelium of the daughter cyst, but CK19 was positive. CK18-mRNA was expressed in all the layers of the lining epithelium, the salivary glands and daughter cysts.

CONCLUSIONSHistological features and CK18 expression may be indicative of the possibility of salivary glandular and odontogenic differentiation.

Adolescent ; Adult ; Epithelium ; pathology ; Female ; Humans ; Keratin-18 ; metabolism ; Keratin-19 ; metabolism ; Male ; Odontogenic Cysts ; metabolism ; pathology

OBJECTIVETo study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium.

METHODSCK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases).

RESULTSThe expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA.

CONCLUSIONSThese results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.

Epithelial Cells ; metabolism ; pathology ; Humans ; Jaw Cysts ; etiology ; metabolism ; pathology ; Keratin-13 ; biosynthesis ; genetics ; Keratin-18 ; biosynthesis ; genetics ; Maxillary Diseases ; etiology ; metabolism ; pathology ; Metaplasia ; metabolism ; pathology ; Postoperative Complications ; RNA, Messenger ; genetics

OBJECTIVETo examine cytokeratin 18(CK18) and it's gene in jaw odontogenic keratocyst (OKC) epithelial lining.

METHODSThe epithelial linings of 32 cases were subject to monoclonal antibody immunohistochemical staining for CK18, CK8 and CK19. RT-PCR and in situ hybridization for CK18 mRNA were conducted in 12 of 32 cases in keratocyst epithelial cell linings.

RESULTSIn 17 cases, CK18 were observed in keratinized surface layers, though weakly positive. In 27 cases, CK18 were positive in the granular cell layers. CK18 were also positive in the spinous cell layers in 14 cases. In all cases, CK18 was negative in basal cell layers. By RT-PCR, 4 cases expressed CK18 strongly, 8 cases weakly. By in situ hybridization, 8 cases expressed CK18 mRNA positively in both spinous and granular cell layers, and 4 cases positively in basal and keratinized cell layers. CK8 were expressed in basal cell layers of keratocyst epithelial linings. In 23 cases, CK19 were expressed in surface cell layers of keratocyst epithelial linings.

CONCLUSIONThe expression of CK18 in keratocyst epithelial linings transfers from basal cell layer to spinous layer. The expression of CK18 immunohistochemical staining and CK18 mRNA in situ hybridization are different, which shows CK18 might be related to proliferation of OKC epithelial linings. That suggests the existence of regulation of CK18 and CK18 mRNA expression.

Epithelial Cells ; Humans ; In Situ Hybridization ; Keratin-18 ; Keratins ; Odontogenic Cysts ; RNA, Messenger

Objective To analyze the clinical characteristics ,causes of death and their changes of hospitalized neonates so as to provide theoretical basis for improving the level of intensive medical care and reduce neonatal mortality .Methods The clinical data of 108 neonates who died between January 2012 and December 2016 were collected .We compared the mortality rate of neonates with different gestational age ,birth weight ,sex ,family background and abnormal high-risk pregnancy .The causes of death and death rate were analyzed .Results Among the 8869 hospitalized neonates ,108 died and the mortality rate of the neonates was 1 .22% .The avoidable mortality rate of the neonates was 0 .86% and the avoidable mortality ratio was 71 .29% .Infectious diseases remained to be the leading cause of neonatal death in hospitals . The top five most common causes of death in our hospitalized neonates were infectious diseases ,respiratory diseases ,asphyxia ,congenital malformations ,and genetic metabolic diseases .The three most common causes of death in full-term infants were infectious diseases ,genetic metabolic diseases ,and asphyxia . The three most common causes of death in preterm infants were infectious diseases , respiratory diseases ,and asphyxia .The neonatal mortality rate in our hospital decreased from 2 .02% in 2012 to 1 .09% in 2016 .Sepsis was the leading cause of death between 2012 and 2015 and dropped to the third place in 2016 . Respiratory diseases were the leading cause of death in 2016 . Asphyxia was the second cause of death in 2016 . Congenital malformations dropped from the third cause of death to the fifth .Conclusion In recent years ,thetreatment of neonates has improved and mortality rate of hospitalized neonates is gradually decreased .Controlling infectious diseases should be the primary measure to reduce the avoidable mortality in hospitalized neonates .

OBJECTIVETo evaluate the effectiveness and safety of subcutaneous low molecular weight heparin (LMWH) used in acute management of patients with non-ST segment elevation acute coronary syndrome (ACS).

METHODSA total of 102 patients with non-ST segment elevation ACS were treated for at least 48 hours ( > or =5 times) with subcutaneous nadroparin (1 mg/kg each 12 hours). All 102 patients underwent coronary angiographies (CAG) within 8 hours after LMWH injection, followed by immediate percutaneous coronary intervention (PCI).

RESULTSAnti-Xa activity at the time of catheterization was (0.62 +/- 0.18) IU/ml, and 90% of the patients had anti-Xa activity > 0.5 IU/ml. No death, myocardial infarction relapse or emergent revascularization occurred after PCI. Thrombosis and/or embolism occurred in 2 patients (3.5%) during PCI. Mild hemorrhage was observed in 4 patients (3.9%) of PCI group and in 2 patients (4.4%) in CAG group. No major hemorrhage occurred.

CONCLUSIONPCI within 8-12 hours of the last dose after > or =48 hours nadroparin subcutaneous injection seems to be effective and safe.

Acute Coronary Syndrome ; blood ; therapy ; Angioplasty, Balloon ; Anticoagulants ; adverse effects ; therapeutic use ; Factor Xa Inhibitors ; Humans ; Nadroparin ; adverse effects ; therapeutic use

BACKGROUNDIslet transplantation represents an ideal therapeutic approach for treatment of type 1 diabetes but islet function and regeneration may be influenced by necrosis or apoptosis induced by oxidative stress and other insults. Heme oxygenase-1 (HO-1) is the rate-limiting enzyme in the catabolism of heme into biliverdin, releasing free iron and carbon monoxide. It has also been reported to be an antioxidant enzyme which can improve the function of grafted islets by cytoprotection via free radical scavenging and apoptosis prevention. In the present study, we investigated whether transduction of HO-1 genes into human islets with an adenovirus vector has cytoprotective action on islets cultured in vitro and discuss this method of gene therapy for clinical islet transplantation.

METHODSCadaveric pancreatic islets were isolated and purified in vitro. Transduction efficiency of islets was determined by infecting islets with adenovirus vector containing the enhanced green fluorescent protein gene (Ad-EGFP) at multiplicities of infection (MOI) of 2, 5, 10, or 20. Newly isolated islets were divided into three groups: EGFP group, islets transduced with Ad-EGFP using MOI = 20; HO-1 group, transduced with adenovirus vectors containing the human HO-1 gene using MOI = 20; and control group, mock transduced islets. Insulin release after glucose stimulation of the cell lines was determined by a radioimmunoassay kit and the stimulation index was calculated. Flow cytometry was used to detect apoptotic cells in the HO-1 group and in the control group after induction by recombinant human tumor necrosis factor-alpha (rTNFalpha) and cycloheximide (CHX) for 48 hours.

RESULTSAdenovirus vectors have a high efficiency of gene transduction into adult islet cells. Transduction of islets with the Ad-EGFP was most successful at MOI 20, at which MOI fluorescence was very intense on day 7 after transduction and EGFP was expressed in cultured islet cells for more than four weeks in vitro. The insulin release in the control group was (182.36 +/- 58.96) mIU/L after stimulation by high glucose media (16.7 mmol/L), while insulin release from the HO-1 group and the EGFP group were (270.09 +/- 89.37) mIU/L and (175.95 +/- 75.05) mIU/L respectively. Compared to the control group and the EGFP group, insulin release in the HO-1 group increased significantly (P < 0.05). After treatment with rTNFalpha and CHX the apoptotic ratio of islet cells was (63.09 +/- 10.86)% in the HO-1 group, significantly lower than (90.86 +/- 11.25)% in the control group (P < 0.05).

CONCLUSIONSTransduction of human islets with Ad-HO-1 can protect against TNF-alpha and CHX mediated cytotoxicity. The HO-1 gene also appears to facilitate insulin release from human islets. Transduction of donor islets with the adenovirus vector containing an HO-1 gene might have potential value in clinical islet transplantation.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cycloheximide ; pharmacology ; Cytoprotection ; Genetic Therapy ; Heme Oxygenase-1 ; genetics ; physiology ; Humans ; Insulin ; secretion ; Islets of Langerhans ; physiology ; Transduction, Genetic ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the association between G894T polymorphism and essential hypertension (EH) in Uygur population in Xinjiang province.

METHODSIn this case-control study, G894T genotypes in 375 hypertension patients (EH group) and 414 normotensive control subjects (NT group) from the rural area of Tuluafan of Xinjiang was investigated by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.

RESULTS(1) Significant differences were found in GG, GT and TT frequencies of G894T genotypes between the EH and NT groups (56.5%, 28.3%, 15.2% in EH group and 65.9%, 22.5%, 11.6% in NT group, OR = 2.97, 95% CI 1.393 - 6.358). T allele frequencies were significantly higher in EH group (29.33%) than that in NT group (22.83%, P < 0.05). (2) SBP, DBP in patients with T allele of eNOS gene [(171.36 +/- 22.30) mm Hg and (103.63 +/- 13.22) mm Hg] were significantly higher than that of GG genotype [(158.07 +/- 20.850) mm Hg and (89.90 +/- 10.39) mm Hg] (P < 0.01).

CONCLUSIONSeNOS gene exon7 G894T polymorphism might relate to essential hypertension in Uygur population in Xinjiang province.

Aged ; Alleles ; Case-Control Studies ; China ; epidemiology ; Exons ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; epidemiology ; ethnology ; genetics ; Male ; Middle Aged ; Nitric Oxide Synthase Type III ; genetics ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide