AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Objective To explore the association between the infection of human cytomegalovirus(HCMV),herpes simplex virus typeⅠ(HSV-Ⅰ),HSV-Ⅱ,toxoplasma(TOX) and serum hepatitis B virus(HBV)these 5 pathogens and low body weight and pneumonia,and explore the clinical value of nested polymerase chain reaction (PCR) examining newborns infected with pathogens.Methods Forty-six newborn infants with low weight and 66 newborn infants with pneumonia were selected.And 1 mL pripheral blood of every newborn infant was drawn.Classic phenol-chloroform-isoamyl alcohol-protease digested,after neonatal serum extraction of DNA in peripheral blood through 2 pairs of pri-mers,the outer primer amplified larger DNA fragments and the inner primer amplified small fragments,in the amplified products.HCMV,HSV-Ⅰ,HSV-Ⅱ,TOX and HBV of the viral DNA in highly conservative district to design primer respectively and amplify its viral DNA,nested PCR was used to detect of these pathogens DNA in infants of low body weight and pneumonia,and to detect positive rate of infection.Screening for birth defects in infants in these virus infection.SPSS 10.0 software was used to analyze the relationship between infection of 5 pathogens.Results The infective rate of HCMV in 46 infants with low body quality was 91.3%,the infective rate of HSV-Ⅰwas 8.7%,the infective rate of HSV-Ⅱwas 15.2%,the infective rate of TOX was 8.7%,and the infective rate of HBV was 15.2%.Among 66 infants with pneumonia,the infective rate of HCMV was 83.3%,the infective rate of HSV-Ⅰwas 6.1%,the infective rate of HSV-Ⅱwas 16.7%,the infective rate of TOX was 6.1%,and the infective rate of HBV was 7.6%.The infective rate of HCMV was higher than that of other 4 pathogens,these infection rates were different statistically in these 5 kinds of pathogens(Pa=0).Conclusions Five kinds of pathogens both low pathosens screening is necessary newborns infants with low body weight and pneumonia,and for the early diagnosis and prevention of these pathogens.

Adult ; Aged ; Aged, 80 and over ; Carcinoma in Situ ; virology ; Carcinoma, Squamous Cell ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; Microarray Analysis ; methods ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; diagnosis ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Uterine Cervical Neoplasms ; virology

BACKGROUNDSeveral studies suggest that cyclooxygenase-2 (COX-2) contributes to the delayed progression of ischemic brain damage. This study was designed to investigate whether COX-2 inhibition with parecoxib reduces focal cerebral ischemia/reperfusion injury in rats.

METHODSNinety male Sprague-Dawley rats were randomly assigned to three groups: the sham group, ischemia/reperfusion (I/R) group and parecoxib group. The parecoxib group received 4 mg/kg of parecoxib intravenously via the vena dorsalis penis 15 minutes before ischemia and again at 12 hours after ischemia. The neurological deficit scores (NDSs) were evaluated at 24 and 72 hours after reperfusion. The rats then were euthanized. Brains were removed and processed for hematoxylin and eosin staining, Nissl staining, and measurements of high mobility group Box 1 protein (HMGB1) and tumor necrosis factor-α (TNF-α) levels. Infarct volume was assessed with 2,3,5-triphenyltetrazolium chloride (TTC) staining.

RESULTSThe rats in the I/R group had lower NDSs (P < 0.05), larger infarct volume (P < 0.05), lower HMGB1 levels (P < 0.05), and higher TNF-α levels (P < 0.05) compared with those in the sham group. Parecoxib administration significantly improved NDSs, reduced infarct volume, and decreased HMGB1 and TNF-α levels (P < 0.05).

CONCLUSIONSPretreatment with intravenous parecoxib was neuroprotective. Its effects may be associated with the attenuation of inflammatory reaction and the inhibition of inflammatory mediators.

Animals ; Blotting, Western ; Brain Ischemia ; drug therapy ; metabolism ; prevention & control ; Injections, Intravenous ; Isoxazoles ; administration & dosage ; therapeutic use ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; metabolism ; prevention & control

OBJECTIVETo investigate the genetic polymorphisms of 11 Y-chromosomal short tandem repeats (Y-STR) loci in 484 male individuals from two minority populations, the Hui and Xibe, of Liaoning province, and to evaluate their forensic application values and genetic relationships with other 15 populations of China.

METHODSEleven Y-STR loci in all samples were amplified with PowerPlex Y System, and the PCR products were analyzed by 310 Genetic Analyzer. Cluster analysis and neighbor-joining tree were applied to show the genetic distance among the populations.

RESULTSIn Hui people, 187 haplotypes were identified, and the overall haplotype diversity value was 0.9990. The gene diversity values (GD) for each locus ranged from 0.4783(DYS437) to 0.9679(DYS385a/b); In Xibe people, 237 haplotypes were identified, and the overall haplotype diversity value was 0.9984. The GD value for each locus ranged from 0.3618(DYS391) to 0.9686(DYS385a/b). Comparing with 15 reference populations, the genetic distance between the Hui and Xibe was the nearest (0.0257), and that between the Hui and Yi was the farthest (0.1046), while the genetic distance between Xibe and Korean was also the farthest (0.0978). The NJ tree was similar to the results of clustering analysis and all the 17 populations were clustered into 3 groups.

CONCLUSIONThe genetic distribution of the 11 Y-STR loci in Liaoning Hui and Xibe ethnic groups showed favorable polymorphisms, therefore are suitable for forensic identification and paternity testing in the local area. The study of haplotype diversity among different populations is useful in understanding their origins, migrations and their relationships.

Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, Y ; Ethnic Groups ; classification ; genetics ; Genetics, Population ; Haplotypes ; Humans ; Male ; Minority Groups ; Phylogeny ; Polymorphism, Genetic ; Tandem Repeat Sequences ; genetics

OBJECTIVETo investigate the value of using an AMACR/34betaE12/p63 cocktail and double-staining for the diagnosis of small focal protatic carcinoma and precarcinomatous lesions.

METHODSA total of 130 consecutive cases were examined over a 3-month period, including 105 prostate needle biopsy samples, 6 radical prostatectomy specimens and 19 benign prostatic hyperplasia specimens which were excised transurethra or above pubis. 262 paraffin blocks of all the 1030 ones were stained with hematoxylin and eosin and by immunostains for AMACR, 34betaE12, p63, and an antibody cocktail comprising all the three with double-chromogen reaction. The diagnoses were then made according to the immunostaining, HE staining and clinical information.

RESULTSIn the sections stained by the 3-antibody cocktail, blue-black cytoplasmic staining was observed in the epithelial cells of prostatic carcinoma and high-grade prostatic intraepithelial neoplasia (HGPIN) the basal cells of benign glands were stained red. There were no red basal cells around the blue-black glandular epithelium of carcinoma, but discontinuous or consecutive red basal cells were present around the blue-black glandular epithelium of HGPIN. Prostatic carcinoma was found in 214 paraffin blocks (82%), including 31 small focal carcinoma. HGPIN were observed in 64 paraffin blocks (24%), including focal HGPIN and small gland alveolus HGPIN. AAH was found in one block. No benign glands were simultaneously positive for AMACR and negative for basal cell markers.

CONCLUSIONInmmunohistochemistry studies using a 3-antibody cocktail and double staining can improve the detection rate of small focal prostatic carcinoma and HGPIN.

Biomarkers, Tumor ; analysis ; Humans ; Immunohistochemistry ; methods ; Keratins ; analysis ; Male ; Predictive Value of Tests ; Prostatic Intraepithelial Neoplasia ; diagnosis ; metabolism ; Prostatic Neoplasms ; diagnosis ; metabolism ; Racemases and Epimerases ; analysis ; Staining and Labeling ; methods ; Trans-Activators ; analysis ; Transcription Factors ; Tumor Suppressor Proteins ; analysis

Objective To explore the relationship between the volatile components in Angelicae Sinensis Radix from different regions of Gansu Province and its growing environment with metabolomics based on GC-MS. Methods The GC-MS method was used for detecting the volatile components in Angelicae Sinensis Radix from 31 different regions in Gansu province, and principal component analysis (PCA) and partial least squares (PLS) methods were used for analyzing and evaluating its relationship with the growing environment. Results The results of PCA showed that the volatile components in Angelicae Sinensis Radix from different regions in Gansu province were related to the altitude and the soil types. The PLS method could divide 31 samples of Angelicae Sinensis Radix from different regions in Gansu Province into three groups according to the difference of altitude. There were significant differences in the volatile components in the samples taken at different altitude regions. After analyzing linear loading plots from PCA and PLS, 11 charateristic components were screened out, including 7 compounds were identified by the retrieval of NIST11 database. Conclusion The volatile components in Angelicae Sinensis Radix from different regions in Gansu Province are closely related to the altitude and the soil type.

Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg^-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between C_max and AUC_0-5d in the two groups. The liquid preparation was the reference preparation, with C_max ratio of 101.6% and AUC_0-5d ratio of 101.9%, the 90% confidence interval of C_max was 79.42%-129.92%, and the 90% confidence interval of AUC_0-5d was 85.72%-121.18%. These results suggested that different dosage forms of MIL50 had certain differences in the changes of blood drug concentration in rhesus monkeys.

Objective To investigate the protective mechanism of protein kinase C(PKC)and calcium sensing receptor(CaR)in ischemia preconditioned rat hearts.Methods Using cell culture method,in vitro cultured inhibitor(IPC+CaRI).Apoptosis was detected using TUNEL and Hoechst33342 cell viability was detected by MTT,the protein expression of easpase-12,calpain and CaR in endochylema were detected using Wedtetm blot.ResultsIn I/R group nucleus was shrank,big blue,chromatin concentrated,apoptotle body appeared.Other groups haddifferent fluorescence intensity varying degree,IPC+PKCI+CaRS group had more big blue nucleus.Myocardialcell viability and apoptotic rate,I/R group[(62.99±0.65)%,(19.13±0.87)%],IPC group[(78.67±0.37)%,(14.21±0.74)%],IPC+PKCI group[(71.09±0.52)%,(20.46±0.81)%],IPC+PKCI+CaRS group(66.10±0.75)%,(24.89±1.43)%],IPC+CaRS group[(69.56±0.44)%,(21.64±0.77)%],IPC+CaRI group(85.81±0.60)%,(13.12±0.69)%],all had a difference(P＜0.05 or＜0.01)compared with C group[(100.00)%,(6.02±0. 31)%].Western blot identified that CaR expression in IPC+PKCI and IPC+CaRS,IPC+PKCI+CaRS groupswas more than that in IPC and IPC+CaRI groups;easpase-12 had more active fragment(60×103)in I/R,IPC+CaRS,IPC+PKCI+CaRS groups;ealpain expressions in I/R,IPC,IPC+PKCI,IPC+PKCI+CaRS,IPC+CaRSgroups were higher than those in C and IPC+CaRI,I/R group was the highest one,C group the second,IPC+CaRI the third.Conclusion The interaction of PKC and CaR can reduce the intracellular Ca2+ from sarcoplasmicreticulum thus provide a protection.

Objective To evaluate the causes of DR waste film rates in different characteristic cases,and provide basis for quality control.Methods DR waste film rates of 98 503 cases from PACS were analyzed.All cases were grouped according to gender,age,X ray projection direction and projection position.The differences of DR waste film rates between the groups were compared.Results DR waste film rate in 98 503 cases was 4.89％ and factors that caused waste film were incorrect position,foreign materials,motion artifacts,machine problems and inappropriate X ray dosage.DR waste film rate was affected by gender,age,X ray projection direction,but not affected by projection position.Conclusion Analysis of DR waste film rate in different characteristic cases could provide basis for the quality control to reduce the DR waste film rate.