Objective To explore the clinical application and outlook of anterior segment observation with Stratus optical coherence tomography(Stratus OCT).Design Prospective case series.Participants 56 eyes in 38 patients with primary angle closure glaucoma (PACG).Methods Morphological changes of anterior chamber angle (ACA) were observed with Stratus OCT before and 4 weeks after laser iridectomy.Opening status of ACA were described and measured with Photoshop software.Main Outcome Measures Opening degree of ACA and ratio of anterior chamber depth to thickness of cornea.Results To a great deal of extent,Stratus OCT could be used to observe the appearance of ACA. Clear images of ACA could be acquired.The ACA before laser was (15.67?5.33) degree and significantly reopened to(26.56?8.17) degree after laser iridectomy (P=0.000).The ratio of anterior chamber depth to thickness of cornea was also changed from (0.39?0.13) pre-treatment to (0.89?0.32) pest-treatment(P=0.000).Conclusions Stratus OCT could be applied to observe the changes of opening degree of ACA easily.The standard of quantization should be consummated in the future.

Animals ; Antioxidants ; pharmacology ; Carbon Tetrachloride Poisoning ; drug therapy ; Chemical and Drug Induced Liver Injury ; drug therapy ; etiology ; Enzyme Inhibitors ; pharmacology ; Male ; Mice ; Protective Agents ; pharmacology ; Ribonucleases ; antagonists & inhibitors

Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of Ca2+, Mg2+, K+, and HCO3 - in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.
ATP-Binding Cassette Transporters/*chemistry/*genetics/metabolism ; Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Calcium/metabolism ; *Cloning, Molecular ; Cryptosporidiosis/parasitology ; Cryptosporidium/chemistry/genetics/*metabolism ; Humans ; Iron/metabolism ; Mice ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/*chemistry/*genetics/metabolism ; Sequence Alignment

Advanced technologies are used to clarify the meridian tropism theory of traditional Chinese medicine is an important part of theoretical studies of traditional Chinese medicine. In this article, modern pharmacokinetic method was used to investigate tissue distribution characteristics of psoralen and isopsoralen of Psoraleae Fructus decoction in rats, in order to provide research ideas and experimental basis for the meridian tropism theory. In this study, various tissue samples such as heart, liver, spleen, lung, kidney, brain and spermary were collected at different times after oral administration with FP decoction, in order to determine concentration of psoralen and isopsoralen by HPLC. Pharmacokinetic parameters were calculated by DAS 2.0 software. The study results showed that HPLC indexes of psoralen and isopsoralen in various tissues of rats met the determination requirements of biological samples. Both components were distributed in all of the tissues, with AUC(0-t) order of liver > lung approximately kidney > heart > brain approximately spleen > spermary. There was significant difference between liver, kidney, lung and other tissues (P < 0.05). MRT(0-t) of both psoralen and isopsoralen were about 10 h. Therefore, psoralen and isopsoralen showed stronger targeting selection in liver, kidney and lung.
Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Fruit ; chemistry ; Furocoumarins ; pharmacokinetics ; Kinetics ; Male ; Plant Extracts ; administration & dosage ; chemistry ; pharmacokinetics ; Psoralea ; chemistry ; Rats ; Tissue Distribution

This study was aimed to propagate and identify the prdm1 gene-knockout mice, so as to lay the foundation for studying Blimp-1 protein. Two kinds of transgenic homozygous mice with B6.prdm1(flox/flox) and B6.Lck-Cre were feed and propagated; after successful propagating, the first passage mice were obtained; after the first passage mice were copulated once again, the genotypes were obtained as follows: B6. prdm1(wild/wild). Lck-Cre, B6. prdm1(wild/wild), B6.prdm1(flox/flox). Lck-Cre, B6.prdm1(flox/wild). Lck-Cre, B6.prdm1(flox/flox), B6. prdm1(flox/wild). The genomic DNA of second passage mice was extracted, the Cre and loxp gene fragments were amplified by PCR, then the size of Cre and loxp genomic DNA were detected by agarose gel electrophoresis. The mice with B6.prdm1(flow/flox). Lek-Cre were used as conditionally prdm1-knockout mice, B6.prdm1(flox/wild). Lck-Cre mice, B6.prdm1(flox/flox) and B6 mice were used as controls. The spleen T lymphocytes and B lymphocytes were sorted by using magnetic beads, the blimp-1 target protein was identified by Western blot. The results showed that the two transgenic homozygous mice had the ability to reproduce, and the separation ratio of second passage mice generated from propagation of their offspring cach other meet Mendelian laws, and the prdm1 gene-knockout mice also could successfully obtained. It is concluded that the application of Cre-loxp system may successfully obtain plentiful prdm1 gene-knockout mice.
Animals ; Genotype ; Mice ; Mice, Inbred C57BL ; genetics ; Mice, Knockout ; genetics ; Reproduction ; Transcription Factors ; genetics

In order to study the distribution of virulence genes of Listeria monocytogenes (Lm) in Hebei Province,29 virulence genes of Lm,including Listeria monocytogenes pathogenicity islands Ⅰ (LIPI-Ⅰ:prfA,plcA,plcB,hlyA,mpl and actA),10 internalins genes (inlA,inlB,inlC,inlD,inlE,inlF,inlG,inlH /C2,inlI and inlJ) and the other 13 virulence-associated genes (bsh,srtA,iap,sigB,virR,mprF,dltA,dltB,dltC,dltD,srtB,fbpA and hpt) were detected by PCR.Results showed that in the 91 Lm strains,the detection rate of 23 virulence genes were 100％.The 29 virulence genes of 26 Lm strains were all detected,and 65 Lm strains had different deletion of 6 virulence genes inlD,inlF,inlG,inlH /C2,inlJ and mpl.The deletion rate of inlG and inlF were 60.44％ and 54.95％,respectively,following by mpl gene,with a deletion rate of 19.78％.According to the absence of virulence genes,91 strains could be divided into 10 subtypes,and the dominant virulence subtypes was type Ⅰ with all 23 virulence genes.The deletion rate of virulent genes in Shijiazhuang was higher than that in northern Hebei.It is suggested that the rate of virulence gene of food-borne Lm in Hebei Province is high,and the virulence gene deletion patterns has diversity and regional differences.

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.
Apoptosis Regulatory Proteins ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; Transfection ; U937 Cells

OBJECTIVETo determine the diagnostic value of B72.3, BerEP4 and calretinin in differentiating metastatic carcinoma cells from reactive mesothelial cells (RMC) in serous effusions by using immunocytochemical method (ICC), and to investigate the feasibility of ThinPrep (TP) preparation for ICC.

METHODSOne hundred fifty eight serous effusion specimens were examined by ICC on cell block (CB) sections (CB-ICC) using antibodies against of B72.3, BerEP4 and calretinin. Fourty-nine of the samples, ICC on ThinPrep slides (TP-ICC) and CB-ICC were performed concurrently.

RESULTSThe sensitivities of B72.3 and Ber-EP4 for detecting carcimoma cells were 76.9% and 69.2% respectively, and when combined the sensitivity was increased to 89.7%. The sensitivity and specificity of Calretinin for detecting mesothelial cells were 90.9% and 87.2% respectively. The sensitivity of B72.3 in differentiating cancer cells from reactive mesothelial cells by CB-ICC and TP-ICC was 78.9% and 68.4%. It was 78.9% and 68.4% of BerEP4 respectively. No statistical significance was observed between CB-ICC and TP-ICC in differentiating metastatic carcinoma cells from reactive mesothelial cells.

CONCLUSIONThe combination of antibodies of B72.3, Ber-EP4 and calretinin is quite helpful as an auxiliary in differentiating metastatic carcinoma cells from reactive mesothelial cells. ThinPrep preparation slides may effectively replace the cell block sections for ICC in differential diagnosis of serous effusions.

Antibodies, Monoclonal ; Antibodies, Neoplasm ; Ascitic Fluid ; metabolism ; pathology ; Calbindin 2 ; Cytodiagnosis ; Diagnosis, Differential ; Humans ; Pericardial Effusion ; diagnosis ; pathology ; Pleural Effusion, Malignant ; diagnosis ; pathology ; S100 Calcium Binding Protein G

In order to explore the transfection and expression of hRI gene on human umbilical blood stem cells, and observe it's effect on the tumor growth. After enriching human umbilical cord blood CD34+ cells with a high-gradient magnetic cell sorting system (MACS), transfected them with supernatant of retrovirus containing human Ribonuclease inhibitor (hRI) cDNA. Hematopoietic progenitor clonogenic assay and PCR were used to evaluate transfection efficiency, and Western-blot and immune fluorescence were used to evaluate the expression quantity of hRI gene after transfection. Observe the effect of RI on the growth of melonoma in B16C57BL mice. The results showed that human umbilical blood CD34+ cells were highly purified by MACS, which made the purity of human umbilical blood CD34+ cells average 96.15%. hRI can be transfected on umbilical blood CD34+ cells, and the transfection efficiency was 35%. The positive expression of hRI gene on transfected CD34+ cells is identified by Western-blot and immune fluorescence assay. Mice injected with transfected CD34+ cells show a significant restraint of the tumor growth, a lower efficiency of tumor formation, a lower weight of the tumor and a longer incubation period of tumor formation with respect to the control groups. The results demonstrated the capacity of RI to inhibited the tumor growth by blocking the vasculature in tumor.
Animals ; Antigens, CD34 ; metabolism ; Cell Proliferation ; drug effects ; Fetal Blood ; cytology ; Genetic Therapy ; Hematopoietic Stem Cells ; metabolism ; Humans ; Melanoma ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Transfection ; Tumor Cells, Cultured

To analyze the metabolites of Chenxiang Huaqi pill in rats by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). The separation was performed on Phenomenex Kinetex C₁₈ column, with the acetonitrile -0.1% formic acid as the mobile phase for gradient elution at a flow rate of 0.8 mL·min⁻¹. The data were collected by the positive ion mode of ESI source. The plasma and urine total ion chromatograms of the rats in blank group and treatment group were used to analyze the targeted ion chromatograms. The results showed that 24 compounds were detected in the plasma and urine, including 5 prototype components and 19 metabolites. The major metabolic pathways included hydration, glucuronidation, demethylation, hydrolysis, hydroxylation and sulfation. The method was rapid, simple and sensitive, and can be used to rapidly identify the metabolites of Chenxiang Huaqi pill that can be absorbed in rats, providing a reference for the study of the absorption and metabolism mechanism of Chenxiang Huaqi pill .
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; metabolism ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Urine ; chemistry