DNA, Fungal ; isolation & purification ; Humans ; Mass Screening ; Occupational Diseases ; diagnosis ; microbiology ; Polymerase Chain Reaction ; Skin ; microbiology ; Tinea ; diagnosis ; microbiology

ADAM Proteins ; genetics ; ADAM17 Protein ; Animals ; Gene Library ; Genetic Vectors ; Male ; Mice ; Two-Hybrid System Techniques

Objective To examine the application value of reticulocyte hemoglobin content (CHr)for diagnosing iron deficiency in premenopausal women.Methods The levels of CHr,hemoglobin (Hb), mean cellular volume(MCV),red cell distribution width (RDW) were measured on the ADVIA 120 (Bayer Diagnostics) automated hematology analyzer.Transferrin saturation (TS) and ferritin (SF) were measured on chemistry analyzer.Results CHr in iron deficiency without anemia group were significantly lower than that in the healthy control group (P＜0.01)and significantly higher than that in iron deficiency anemia group(P＜0.01).CHr in anemia of chronic disease group were significantly higher than that in iron deficiency anemia group(P＜0.01).Receiver operator characteristic curve (ROC) analysis in diagnosis of iron dificiency without anemia demonstrated that the area under the curve for CHr,SF,RDW,MCV,Hb were 0.872,0.798,0.721,0.713,0.677,respectively (P＜0.01).So CHr has a better overall sensitivity than SF,Hb,MCV and RDW in the diagnosis of iron deficiency without anemia.ROC also showed that the area under the curve for Hb,RDW,CHr,SF and MCV was 1.000,0.969,0.958,0.953 and 0.926,respectively (P＜0.01) in iron deficiency anemia.Conclusion CHr is the early and sensitive predictor of iron deficiency in premenopausal women,especially for the diagnosis of iron deficiency without anemia.

Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P＜0.01),which was contrary to those transfected with antisense MTHFR(P＜0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.

In this study, the effect of heparin-derived oligosaccharide (HDO) on platelet-derived growth factor (PDGF) induced vascular smooth muscle cells (VSMCs) proliferation and the related signal transduction mechanisms were investigated. MTT assays were used to measure VSMCs proliferation. Cell cycle distribution was analyzed by flow cytometry. The level of key regulatory proteins in PKC, MAPK and Akt/PI3K pathways were determined by RT-PCR, Western blot and immunocytochemical methods. Meanwhile, mRNA expressions of some proto-oncogenes were assayed by RT-PCR method. Our data showed that HDO (0.01, 0.1 and 1 μmol · L(-1)) inhibited 30 ng · mL(-1) PDGF-induced VSMCs proliferation in a dose-dependent manner, blocked the G1/S transition and inhibited the level of key regulatory proteins and some proto-oncogenes (P < 0.05). The results showed that HDO may decrease the key regulatory proteins expression, hence suppress the transcription of proto-oncogene and G1/S transition, finally inhibiting VSMCs proliferation.
Cell Cycle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Heparin ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Oligosaccharides ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Signal Transduction

OBJECTIVETo prepare monoclonal antibodies of mice anti-human sperm protein 22 (SP22) and to identify their specificities.

METHODSBALB/c mice were immunized with human SP22, monoclonal antibodies were prepared by hybridoma technique and the sensitivity and specificity of SP22 McAb were investigated by ELISA and Western-blot assay, respectively. The distribution of human SP22 in sperm were shown by immunohistochemistry.

RESULTSThree strains of hybridoma cells were obtained, with the affinity constant (K) of 1.0 x 10(7) L/mol and the titers were 1:10(3) and 1:3,200 in the mixed supernatant of cell cultures and abdominal dropsy, respectively. IgG isotype of the antibody was identified as IgG1. Western blot demonstrated that there was a specific recognition between human SP22 and the obtained monoclonal antibody. Immunohistochemistry displayed that human SP22 mainly distributed on the acrosome surface of human sperm.

CONCLUSIONThe monoclonal antibodies of anti-human 22 was prepared by the technique of hybridoma cell has higher titer and specificity, which can combine specially with the SP22 protein on the surface of human sperm.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hybridomas ; secretion ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred BALB C ; Microtubule-Associated Proteins ; immunology ; metabolism ; Spermatozoa ; metabolism

Objective To explore main technical points and clinical efficacy of individualized stepwise multiple embolization treatment for refractory hemoptysis. Methods To retrospectively analyze materials of 103 patients treated by individualized stepwise multiple embolization. According to disease categories, individualized stepwise multiple embolization treatment with polyvinyl alcohol and loaded sodium alginate microspheres as basic embolization agent were performed, after the type, number, abnormal branches, pulmonary circulation and systematic pulmonary shunt of targeted vessels were confirmed through radiography. To judge short(less than 3 months), medium(3 to 6 months) and long term(more than 6 months) efficacy, resolution of hemoptysis after operation were assessed. To evaluate efficacy of individualized stepwise multiple embolization treatment for refractory hemoptysis, Kaplan-Meier survival curves were used. According to the features of target vessels to supply blood, patients were classified into with SPS and without SPS. By using Log-Rank test, the effective rates of one-year were compared between them. Results Out of 103 patients, 215 target vessels were demonstrated, among which individualized stepwise multiple embolization was for 196 target vessels, peripheral embolization for 8 vessels, and main trunk embolization in 11 patients. The visits after operation were made to 103 patients after 6 to 50 months, with the medium of 21 months. Hemoptysis was instantly resolved in 97.1%(100/103). The effective rates were 94.5%,93.2%, 89.7%,88.9%,85.2%and 76.6%for one, three, six months and one, two and three years after operation. In 103 patients, patients with SPS were 22 and without SPS were 81. One-year effective rates with and without SPS were (69.50 ± 0.11)% and (98.30 ± 0.03)% , respectively (χ2=11.662,P<0.01). Conclusion Individualized stepwise multiple embolization treatment shows excellent short-term and mid-long term efficacy in the treatment of refractory hemoptysis.

Objective To investigate the effects of TLR7 on imiquimod induced apoptosis of THP-1 derived macrophages.Methods Three cell lines ( THP-1 derived macrophages, MDCK cell line and HUVEC cell line) with different capabilities of expressing TLR7 were selected.The survival rates of cells af-ter the treatment with different concentrations of imiquimod were detected by MTT assay.The levels of IL-6 in the supernatants of TLR7 inhibitor chloroquine or TLR7-siRNA treated cells were detected by enzyme-linked immunosorbent assay.The apoptosis of cells was detected by flow cytometry after inhibiting the ex-pression of TLR7.Results Imiquimod induced the apoptosis of THP-1 derived macrophages, MDCK cell lines and HUVEC cell lines.The levels of IL-6 were significantly decreased as the expression of TLR7 was inhibited by treating THP-1 derived macrophages with chloroquine or TLR7-siRNA.Treating THP-1 derived macrophages with chloroquine or TLR7-siRNA did not affect the cell apoptosis induced by imiquimod.Con-clusion Imiquimod could induce the apoptosis of THP-1 derived macrophages through TLR7 independent pathway.

Objective To develop a BP neural network to differentiate between ventricular fibrillation( VF) and non-VF rhythms.Methods Eighteen metrics were extracted from the ECG signals.Each of these metrics respectively characterized each aspect of the signals, such as morphology, gaussianity, spectra, variability, and complexity.These metrics were regarded as the input vector of the BP neural network.After training, a classifier used for VF and non-VF rhythm classification was obtained.Results and Conclusion The constructed BP neural network was tested with the databases of VFDB and CUDB, and the accuracy was 98.61%and 95.37%, respectively.

AIM: To analyze the clinical features of children ocular penetrating trauma,and hope to effectively assist to prevent pediatric ocular trauma METHODS: The data of 145 cases (145 eyes) with ocular penetrating trauma,hospitalized in Xijing Hospital from January 2012 to December 2016,were collected and retrospectively analyzed.All the data of injury factors and environment,age and gender of patients,lesions,treatment and prognosis were detailed studied.RESULTS: In all the 145 pediatric patients with ocular penetrating trauma,accounted for 8.5% of all the ocular trauma patients,there were 95 cases of male,and 50 of female.Penetrating injuries mainly occurred in age of 3-9.The main injuries of ocular perforating in children were scissors,and sharp objects of wooden and iron.The wound was often located in the cornea or the anterior sclera.Traumatic cataract,vitreous hemorrhage and endophthalmitis were the common complications.The visual acuity was severely damaged,and 90 cases (62.1%) of the children recovered better than 0.1 after effective treatment.CONCLUSION: The visual function of pediatric p0atients was seriously threatened after penetrating injuries.However,the damage of vision and the rate of blindness can be reduced effectively after timely and correct diagnosis and treatment.It is the most important that active and effective prevention in keep children away from penetrating injuries.