ObjectiveTo establish a new method for preparing synaptosomes.MethodsDensity gradient centrifugation method was used to isolate synaptosomes of mouse, checking by transmission electron microscopy.ResultsSynaptosomes prepared by this method had intact morphological characteristics, surrounding with a continuous oval-shaped membrane structure, moreover, mitochondrion and lots of synaptic vesicle in them.ConclusionThis method is applicable to establish a rapid, convenient and useful method for preparing synaptosomes.

OBJECTIVETo study the roles of IL-4, IL-5 and IgE in childhood cough variant asthma (CVA).

METHODSThe IL-4 and IL-5 levels in peripheral blood mononuclear cell (PBMC) and the serum IgE levels were determined using ELISA in children with CVA in the acute stage (n=21) and in the convalesce stage (n=9). The samples from 30 children with acute bronchial asthma and from 30 healthy children were used as controls.

RESULTSThe levels of PBMC IL-4 (91.57 +/- 12.19 ng/L) and IL-5 (13.28 +/- 0.31 ng/mL) in children with CVA in the acute stage were significantly higher than those in the convalesce stage (74.68 +/- 11.54 ng/L, 6.53 +/- 0.28 ng/mL) and also higher than those in the healthy controls (70.32 +/- 18.16 ng/L, 5.29 +/- 0.36 ng/mL) (P < 0.01). The levels of serum IgE in children with CVA in the acute stage (279.6 +/- 41.3 KU /L) were strikingly higher than those in the convalesce stage (153.8 +/- 37.5 KU/L) (P < 0.01). The levels of serum IgE in children with CVA either in the acute stage or in the convalesce stage were significantly higher than those in healthy controls (90.6 +/- 44.8 KU /L) (P < 0.01). There were no significant differences in the levels of IL-4, IL-5 and IgE between children with acute CVA and acute asthma.

CONCLUSIONSA combined determination of PBMC IL-4 and IL-5 and serum IgE may be valuable for the diagnosis and the outcome evaluation of CVA. IL-4 and IL-5 may play a role in the pathogenesis of CVA. It is speculated that CVA may have similar pathogenesis to bronchial asthma since acute CVA patients have similar IL-4, IL-5 and IgE levels to children with acute bronchial asthma.

Asthma ; immunology ; Child, Preschool ; Cough ; immunology ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Interleukin-4 ; blood ; physiology ; Interleukin-5 ; blood ; physiology ; Male

OBJECTIVETo evaluate the contribution of individual risk factor to a disease on someone with several risk factors.

METHODSA method based on epidemiological theory and Bayes' theorem was established to measure the contribution of individual risk factor, using the relative risk (RR) or odds ratio (OR) value obtained from population-based cohort studies or meta-analysis.

RESULTSThe proportional contribution for individual risk factor to disease in one person can be measured or estimated.

CONCLUSIONThis method can be applied to risk assessment in a patient with more than one risk factor, and the results also contribute to our etiological study and clinical decision-making strategy.

Bayes Theorem ; Data Interpretation, Statistical ; Disease ; etiology ; Epidemiologic Methods ; Epidemiologic Studies ; Humans ; Proportional Hazards Models ; Risk Assessment ; Risk Factors

OBJECTIVETo investigate the MR image features of TMJ coronal structures.

METHODS28 TMJs from 15 cadavers dipped in formalin over 1 year were included. Their mandibles were fixed at intercuspal position when TMJs oblique coronal MR scan was taken. Baseline was draw on the oblique coronal imaging along the ramus of mandible through its center. The corresponding dimensions and areas were measured with electronic ruler.

RESULTSThe dimensions were in accordance from large to small as: lateral-medial dimension of fossa, lateral-medial dimension of condyle, medial dimension of fossa, medial dimension of condyle, lateral dimension of condyle and lateral dimension of fossa. Three significant co-relative relationship were found: the lateral dimension of condyle with lateral dimension of fossa, the medial dimension of condyle with medial dimension of fossa, and the lateral-medial dimension of condyle with lateral-medial dimension of fossa. CV of lateral dimension of fossa was higher than that of lateral and medial dimension of condyle, and then all the three were higher than other dimensions. The size of the area of joint space shown on oblique coronal imagings was positively related to that of the area of coronal disc imagings.

CONCLUSIONSignificant co-ordinate relationship of condyle and fossa of TMJ coronally was existed, and the condyle position to fossa lateral-medially varied in great.

Cadaver ; Humans ; Magnetic Resonance Imaging ; Temporomandibular Joint ; anatomy & histology

OBJECTIVETo introduce the advantages of the circumcision with a scalpel by inserting a piece of gauze between the inner prepuce and superficial fascia.

METHODSFrom November 2000 to March 2006, 2 100 patients with redundant prepuce, aged 6-78 years, averaging 23, were circumcised with a scalpel by inserting a piece of gauze between the inner prepuce and superficial fascia. Of all the cases, 1 799 (85.7%) were classified as Type I, 237 (11.3%) as Type II and 64 (3.0%) as Type III redundant prepuce. The mean operation time was 20 minutes (15-35 min).

RESULTSThe incisions were all healed in one stage, with good appearance, no infection and no disruption. Bleeding occurred in 12 cases 648 hours after the operation and delayed bleeding in 3 cases.

CONCLUSIONThe circumcision with a scalpel by inserting a piece of gauze between the inner prepuce and superficial fascia, with easier performance, fewer complications and less bleeding, and capable of preserving more and continuous superficial fascia and giving a better appearance, well deserves to be popularized in clinical practice.

Adolescent ; Adult ; Aged ; Child ; Circumcision, Male ; methods ; Humans ; Male ; Middle Aged ; Phimosis ; surgery ; Surgical Sponges

To investigate the effect of hepatitis B virus X protein(HBx) on CtBP-interacting protein(CtIP) which is an important repair factor of DNA double strand break damage in HepG2 cells induced by bleomycin. A HBx stably expressing HepG2 cell line and a control HepG2 cell line with empty vector transfected were established. After the double strand break (DSB) damage occurred, the mRNA and protein levels of CtIP were detected by Real-time PCR and Western blot assay respectively, cell cycle profiles and apoptotic cell death were determined by a flow cytometry, and the position of CtIP in cells was observed by confocal laser scanning microscopy. It showed that HepG2 cells transfected with hepatitis B virus X gene could stably express HBx protein. After being induced by bleomycin, the percentage of apoptotic cell was 16.90%+/-0.89% in HBx stably expressing HepG2 cell line and 15.30%+/-0.86% in control cell line, respectively (q = 2.074, P is more than to 0.05). While the percentage of death cell was 8.71%+/-0.74% in HBx stably expressing HepG2 cell line and 4.90%+/-0.46% in control cell line, respectively (q = 7.126, P is less than to 0.01). The two cell lines manifested the increase of G2/M arrest and significant difference existed between the two cell lines. HBx down regulated the expression levels of CtIP and its mRNA. The CtIP level was 0.66+/-0.04 in HepG2-HBx cell and 0.73+/-0.05 in HepG2-vec cell, respectively (t = 2.314, P is less than to 0.05). The relative mRNA level was 1.00+/-0.06 in HepG2-HBx cell and 1.23+/-0.08 in HepG2-vec cell, respectively (t = 2. 732, P is less than to 0.05). We also found that CtIP was concentrated in the cell nucleus. The research suggests that HBx may affect DNA-repair pathways by disrupting the expression of CtIP.
Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; Real-Time Polymerase Chain Reaction

Objective To discuss the feasibility of Mongolian medicine pulse diagnosis in assessment of the disease situation of acute critically ill patients. Methods The clinical data of acute critically ill patients admitted in the Department of Emergency of Inner Mongolia International Mongolian Medical Hospital from December 2015 to December 2017 were retrospectively analyzed, and the role of Mongolian medicine pulse diagnosis in the evaluation of disease situations of patients with acute critical illness was observed. Results The improved early warning score (MEWS) at admission was carried out for all the patients in the Emergency Department of this hospital, 996 critically ill patients with their MEWS score ≥ 5 scores in accord with the inclusion criteria were allowed to undergo the Mongolian medicine pulse diagnosis, and of them, 654 patients showed specific changes in pulse condition. Shock appeared rapid pulse, microphygmia in pulse diagnosis of Mongolian medicine, severe anemia appeared the slippery pulse and rapid pulse in pulse diagnosis of Mongolian medicine, severe arrhythmia appeared rapid pulse and slow pulse in pulse diagnosis of Mongolian medicine, acute myocardial infarction appeared rapid pulse, weak pulse and slow pulse in pulse diagnosis of Mongolian medicine, insufficiency of heart function appeared rapid pulse and weak pulse in pulse diagnosis of Mongolian medicine, hypertension complicated with cerebral hemorrhage with forceful pulse and pulsus pulse are all suggestive of critical condition. Conclusions The specific changes in pulse condition of patients with acute critical illness in Mongolian medicine are helpful to the assessment of the patient's condition. This method can also be used as one of the tools for rapid assessment of disease situation in critical patients before going to hospital or in the hospital.

[Objective]To study the rare alleles frequencies and sequences of Expressmarker 22 kit in Guangdong Han Population.[Methods]3495 Samples from unrelated individuals in Guangdong Han Population were screened by using AGCU Expressmarker 22 kit(EX22)and ABI 3100 Genetic Analyzer. Then analyzed the frequencies of the off-ladder(OL)alleles and sequenced the rare alleles obtained based on comparison with the STRBase database and litera-ture.[Results]33 off-ladder alleles with 25 rare alleles were found in 10 STR loci,and allele frequencies ranged from 0.0003~0.0046. Sequencing of the11unreported rare OL alleles showed that most of them have incomplete repeats.[Conclusion]Off-ladder alleles especially the rare alleles are helpful to improve the power of discrimination and the power of exclusion and to provide samples which will be added its allele into ladders These OL-alleles will supplement forensic DNA database.

Objective To investigate the protective effect of micro RNA(miR)-98-5p targeting Kruppel-like factor 9(KLF9) against myocardial isehemia-reperfusion(MI/R) injury in rats. Methods Totally 50 rats were randomly divided into sham operation group, model group, miR-98-5p agomir group, agomir-NC group, and miR-98-5p agomir+pcDNA-3. 1-KLF9 group, 10 in each group. MI/R model was established by coronary artery ligation. The pathological changes of myocardial tissue were detected by HE staining. The myocardial apoptosis were detected by TUNEL. The levels of creatine kinase (CK), creatine kinase isoenzymes (CK-MB) and lactate dehydrogenase (LDH) in serum were detected by ELISA. The expression levels of miR-98-5p and KLF9 mRNA were detected by Real-time PCR. The expression of KLF9, Bax and JAK2/STAT3 pathway relative protein were detected by Western blotting. Dual luciferase assay verified the relationship between miR-98-5p and KLF9. Results Compared with the sham operation group, the arrangement of myocardial cells in the model group was disordered, and the myocardial cells appeared necrosis. The apoptosis rate of myocardial cells, serum CK, CK-MB and LDH contents increased, the expression level of miR-98-5p decreased, and the expression levels of KLF9 mRNA and protein, p-JAK2 and p-STAT3 protein increased (P < 0. 05) . After the overexpression of miR-98-5p, the myocardial cells arranged more orderly and the myocardial cell necrosis decreased. The apoptosis rate of myocardial tissue, the contents of CK, CK-MB and LDH in serum and the expression of p-JAK2 and p-STAT3 protein were decreased (P< 0. 05) . The result of dual luciferase assay showed that KLF9 was the target gene of miR-98-5p. The overexpression of KLF9 reversed the effects of miR-98-5p agomir on myocardial injury. Conclusion MiR-98-5p targeting KLF9 can improve the myocardial injury of MI/R rats. The mechanism may be related to the regulation of JAK2/STAT3 signal pathway by miR-98-5p which inhibit myocardial cell apoptosis.

Objective To explore the role of the abnormal expression of miRNAs in the development process of non-small cell lung cancer and the feasibility of ultrasound microbubble-mediated gene therapy after transfecting antisense miRNA-224 and miRNA-122a plasmids into non-small cell lung cancer A549 cells. Methods Antisense miRNA-224 and miRNA-122a plasmids were transfected into non-small cell lung cancer A549 cells on the optimal ultrasound microbubble-mediated condition. We set up a control group. The cell proliferation activity, apoptosis, invasion ability were detected by MTT assay, Annexin V-PE, Transwell invasion experiment and colony formation assay, respectively. Results The expression of miRNA-224 decreased and the expression of miRNA-122a rose after the plasmids of target genes were transfected into non-small cell lung cancer A549 cells, and there were significant differences when compared with those of the control group (P < 0.05). After the plasmids of target genes were transfected into A549 cells, the growth of antisense miRNA-224 and miRNA-122a were inhibited, and the differences were significant as compared with the control group (P < 0.05). Besides, the inhibition of miRNA-122a group was the most significant and there was statistically significant difference as compared with miRNA-224 group (t = −4.694, P = 0.009). After the plasmids of target genes were transfected into A549 cells, the proportion of apoptotic cells increased, the invasive cells were decreased and the clone ability reduced, and also there was a significant difference as compared with those of the control group (P < 0.05). What's more, the apoptotic peak appeared in miRNA-122a group. Its invasion ability decreased most obviously (40.25 ± 3.97/visual field), the number of clone ability was 104.93 ± 4.87 and the inhibitory effect was the most obviously. There was statistically significant difference as compared with other groups (P < 0.05). Conclusions A549 cells transfected by ultrasound microbubble-mediated antisense miRNA-224 and miRNA-122a plasmids possessed good transfection efficiency. The cell growth, invasion and colony-forming abilities of transfected A549 cells were suppressed, which laid a solid foundation for the gene therapy of non-small cell lung cancer.