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 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.

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.

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

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.

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

·AlM: To investigate pathogeny and effects of surgery on paralytic strabismus.
· METHODS: A retrospective study was done in 46 patients with paralytic strabismus who underwent squint correction in our hospital from June 2010 to June 2013. Among 26 horizontal strabismus, the cases of extra rectus palsy was 16, internal rectus palsy was 10.Among all20 vertical strabismus, the cases of superior oblique palsy, superior rectus palsy, inferior rectus palsy, double elevator palsy counted for 7, 8, 2 and 3, respectively. Pathogenesis: trauma was 19 cases, followed by 10 cases that the causes could not be identified.Nine was congen ital paralytic strabismus, 8 o ccurred after nose or brain surgery. The surgery methods included rectus muscle recession, rectus muscle resection, partial rectus muscle transposition, Jensen procedure, inferior oblique myectomy and anterior transposition of inferior oblique. Statistical software SPSS10.0 was used in chi-square test between two groups, while the situation of paralysis eye movements improved by two methods in the horizontal strabismus group was compared with t test.
· RESULTS: Among all horizontal strabismus the rate of cure, improvement and inefficiency was 20 ( 77%) , 5 ( 19%) and 1 ( 4%) , respectively. Among vertical strabismus the ratio of cure, improvement and inefficiency was 15 (75%), 3 (15%) and 2 (10%).There was no significantly difference between the two groups ( P >0.05 ). The movements of paralytic eyes were improved. Two procedures used in horizontal strabismus, can improve paralysis eye movements were 3.76 ±0.91, 3.72 ±0.84mm, with no significant difference (P=0.93) statistically.
· CONCLUSlON: Paralytic strabismus in adults had complicated conditions. Choosing different operation methods in treating paralytic strabismus according to the degree of paralysis can result in satisfactory cosmetically alignment of the eyes and modify head position and diplopia.

Objective To investigate the value of contrast-enhanced ultrasonography (CEUS) in the diagnosis of tuberculous mesenteric lymphadenitis by analyzing its enhancement pattern. Methods The conventional ultrasound and contrast-enhanced ultrasound images of 62 patients with tuberculous mesenteric lymphadenitis confirmed by needle core biopsy or surgery were retrospectively analyzed. The location, size, shape, internal echo and posterior enhancement of mesenteric lymph nodes were recorded. All cases were divided into two groups:the maximum diameter of the lymph node≤20 mm and the maximum diameter of the lymph node >20 mm, and the patterns of enhancement in two groups were analyzed. Results The conventional ultrasound of 62 cases with tuberculous mesenteric lymph nodes showed enlargement. And the echogenicity was hypoechoic or heterogeneity, containing punctate or clusters of calcification in 19 cases (30.6%). After CEUS, there were three forms of enhancements:rim enhancement in 29 cases (46.8%);inhomogeneous enhancement in 21cases (33.9%);non-enhancement in 12 cases (19.3%). Rim enhancement was more common in the≤20 mm group, while inhomogeneous enhancement was more common in the lymph nodes>20 mm. There was statistically significant difference of the enhancement type between the≤20 mm group and the>20 mm group (χ2=6.782, P=0.034). Conclusions Most of tuberculous mesenteric lymph nodes showed rim and inhomogeneous enhancement in CEUS, and the sizes of mesenteric lymph node tuberculosis influenced the CEUS enhancement patterns. CEUS may provide useful information for the diagnosis of the tuberculous mesenteric lymph node.

Objective To observe and comparatively analysize the effect of intraperitoneal injection of hydrogen and hydrogen-rich saline in neonatal rats with hyperoxia-induced lung injury.Methods Sprague-Dawley(SD) newborn rats were randomly divided into six groups(n=10).air group(A),air+ hydrogen-rich saline group(B),air+hydrogen group(C),hyperoxia group(D),hyperoxia+hydrogen-rich saline group (E) and hyperoxia+ hydrogen group(F).The group A,B and C were exposed to air and group D,E and F were exposed to 95 ％ oxygen.The group B and E were intraperitoneally injected with hydrogen-rich saline (10 mL/kg,twice daily),while the groups C and F with hydrogen (10 mL/kg,twice daily).The group A and D were injected with normal saline(10 mL/kg,twice daily).Lung tissue and serum samples were collected on 15 d of experiment.The pathological changes of lung tissue and radiate alveoli count (RAC) were observed by HE staining.The content of HYP in lung tissue was detected by the alkaline hydrolysis method,serum SOD and MDA levels were measured.The expression of α-SMA in lung tissue was detected by immunohistochemistry SP method.Results Compared with the A group,RAC and SOD activities in the D group were significantly decreased,while the HYP and MDA levels and α-SMA expression were significantly increased.Hydrogen intervention could significantly alleviate these changes caused by hyperoxia.while intraperitoneal injection of hydrogen got better effect than intraperitoneal injection of hydrogen-rich saline.Conclusion Hydrogen can extenuate the indexes of hyperoxia-induced lung oxidative damage,impairment development and fibrosis to a certain extent.Intraperitoneal injection of hydrogen has slightly better effect than hydrogen-rich saline.