Objective To explore the simplified method by examining the levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),estradid(E2) and stosterone(T) at different times in the stimutation test of luteinizing hormone-releasing hormone(LHRH).Methods Sixty patients with precocious puberty accepted LHRH stimutation test.The levels of E2,T,FSH,LH before injection and after 30,60,90 minutes were compaired.The levels of stimutation test of LHRH examined with ACS:180 chemiluminesence.Results There were 39 patients in the group of central precocious puberty(CPP).The levels of FSH and LH in CPP group significantly increased after LHRH 30 minutes injection.The ratio of LH/FSH was higher than 1.The peak level of LH was higher than 12 IU/L.There were 21 patients in group of peripheral precocious puberty(PPP).Compared with the results before injection,the levels of LH were similar to the results of 30,60 and 90 minutes after LHRH injection.Compared with the result before injection,the levels of E2 and T were similar to the result of 60 minutes after LHRH injection.The peak levels of LH and FSH in two groups all focused in 30,60 minutes.Conclusions LHRH stimutation test is mainly based on the peak level of LH and the ratio of LH/FSH,the test can be simplified to examine the levels the of LH and FSH before the test and 30,60 minutes after injection as a basis for the clinical diagnosis.

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

Objective To investigate clinical significance of computed tomography (CT) scan in diagnosis of bronchial foreign body in children.Methods Twenty-one suspected children with bronchial foreign body were studied with spiral CT cross-section scan and coronal reconstruction and diagnosis was confirmed with bronchoscopy.Results The foreign body was displayed in all of 21 cases. CT scan showed foreign body was located in right main bronchial 12 cases, right middle bronchial 1 case, right inferior lobar bronchial 2 cases and left main bronchial 6 cases. Foreign bodies were extracted with bronchoscopy.Conclusion CT scan can display and locate accurately foreign body in bronchial of children,and has very important diagnostic value in patients having atypical histories, clinical and radiological findings.

Objective To investigate the effect of angiotensin Ⅱ(Ang Ⅱ)on the expression of integrin-linked kinase(ILK)in rat glomerular mesangial cells.Methods SD rat glomerular mesangial cells were separated and cultured.They were treated with 10-7 mol/L Ang Ⅱ in various time-point(0,12,24,48,72 h),and then cultured rat glomerular mesangial cells were treated 48 h with Ang Ⅱ in various concentration(0,10-11,10-9,10-7,10-5 mol/L).Then total protein expression in rat glomerular mesangial cells stimulated by Ang Ⅱ at every time-point and every concentration-point was extracted.The expression of ILK protein was measured by Western blot.Results 1.Expression of ILK protein in rat glomerular cells stimulated by Ang Ⅱ in various time-point,or various concentration was significantly increased.In group of 10-7 mol/L Ang Ⅱstimulated cells,the protein semiquantity of ILK at 0,12,24,48,72 h respectively was 0.18?0.02,0.37?0.07,0.90?0.10,1.73?0.12,and 1.72?0.13(F=166.78 P

Background Heat shock proteins (HSPs) are highly conserved proteins that are induced in cells when confronted with a wide variety of proteotoxic stresses.HSP27 has a high degree of similarity with α-crystallin protein.The abnormality of HSP27 structure and expression are closely related to the formation of cataracts.Our previous study showed sodium salieylate has the protective effect on H2O2-induced lens damage.Objective This study was to investigate the roles of MAPK signal pathway in sodium salicylate-induced the expression of HSP27 in human lens epithelial cells (LECs) in vitro.Methods Human LECs were incubated in the fresh media containing sodium salicylate at different concentrations (0-55 mmol/L) for different times (1-5 hours) and allowed to be recovered in fresh medium without sodium salicylate for 1-24 hours with or without pretreatment with P38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor ( SP600125). The expressions of P38MAPK, EBK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 were detected by Western blot. HSP27 mRNA was detected by RT-PCR. The expression of HSP27 was also detected by immunohistochemistry. Results There was only weak expression of HSP27 in normal human LECs.After stimulation of 35-55 mmol/L sodium salicylate was removed and human LECs were cultured again for 6 hours,the expression of HSP27 in LECs were significantly increased ( F= 509. 953,P<0. 01). HSP27 was absent expressed in human LECs in 55 mmol/L sodium salicylate stimulation for 1-5 hours groups, but LECs were re-cultured for 3,6 hours after removed the stimulation, the expression of HSP27 was elevated (F = 452. 534, P<0. 01). Activation of P38 M APK occurred after sodium salicylate stimulation 30 minutes and 1 hour ( F = 865.68, P<0. 01). However, ERK 1/2 was expressed after sodium salicylate was eliminated for 1-6 hours ( F = 388.84, P<0. 01). JNK/SAPK was inactived by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059. Conclusion Sodium salicylatc can induce the expression of HSP27 in human (LECs) . The effects are mediated,at least in part ,through the activation of P38MAPK and ERK1/2 signaling pathway .

Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.

Objective To explore the relationship of proteinuria,immunopathogenesis and humoral immunity in children with Henoch-schonlein purpura nephritis(HSPN).Methods Serum IgG,IgA,IgM,C3 and C4 levels were detected with complete-automatic biochemistry analyzer in 47 children with HSPN and 18 heathy children,and the differences in the findings were compared among groups.The levels of IgG,IgA,IgM,C3 and C4 were compared among groups according to the count of proteinuria.The renal tissues were adopteded with immunity in the children with HSPN were taken for biospy examination,and the differences in immunopathalogy were compared among groups with immunof-luorescence assay.Results The IgG,IgA levels in children with HSPN were higher than those of heathy children,on the contrary,the IgM,C3 and C4 levels were lower than those of heathy children(Pa0.05),but the IgM level of the nephritic syndrome group were higher than that of other groups(Pa0.05).Conclusions There is severe disturbance of immunologic function in children with HSPN.There is no connections among serum IgG,IgA,IgM,C3 and C4 levels,proteinuria and renal immunopathlogy.

OBJECTIVETo explore the feasibility and effect of percutanuous cannulated pedicle screw AF system fixation for the treatment of thoracolumbar flexion-distraction fracture without neurologic impairment.

METHODSFrom June 2008 to June 2010,21 patients with thoracolumbar flexion-distraction fracture were treated with percutanuous cannulated pedicle screw fixation. There were 16 males and 5 females with the mean age of 32.7 years ranging from 23 to 55 years. Injured levels 5 cases was in T12, 13 was in L1, 3 was in L2. According to classification of AO, B1 was in 13 cases, B2 was in 8 cases. ASIA grade of all the patients were grade E. The mean operative time, bleeding volume, lengths of stay were evaluated. All the patients took the X-ray and three-dimensional CT reconstruction to observe the fracture healing and to measure the height of the anterior border of fracture vertebral body and the Cobb angle of kyphosis. The visual analogue scales (VAS) were compared preoperation and postoperation. The clinical effects were analyzed according to Oswestry Disability Index (ODI) from 10 aspects, including pain intensity, personal care, lifting, walking, sitting, standing, sleeping, sex life, social life, traveling.

RESULTSAll the operations were successful and the mean time was (109.0+/-16.0) min, blood loss was (90.0+/-15.0) ml, lengths of stay was (7.7+/-2.3) d. No postoperative neurological deficits or wound infection occured. One case occurred subcutaneous fluid of incision and 1 case occurred delayed healing of incision. Fifteen patients were followed up with an average of 16.7 months (12-33 months). The VAS score improved from preoperative 8.3+/-1.7 to 1.8+/-1.2 at final follow up (P<0.05). The ODI of last follow-up was (10.0+/-1.2)%. Three-dimensional CT reconstructions and X-rays showed the height of vertebral body increased from preoperative (54.0+/-17.1)% to (82.7 - 3.5)% at 12 months after operation (P<0.05). Cobb angle of kyphosis changed from preoperative (23.0+/-13.1) to (6.9+/-5.1) degrees at 12 months after operation (P<0.05). The healing time of fracture was (4.7+/-0.3) months. There were no significant loss of vertebral height or kyphosis correction between postoperative 1 months and last follow-up (P>0.05). There were satisfactory fracture reduction and good position of internal fixation,and no loss of reduction within 12 months and no fixation failure in follow-up.

CONCLUSIONThe minimally invasive percutaneous cannulated pedicle screw fixation technique is feasible in treatment of thoracolumbar flexion-distraction fracture without neurologic impairment. This technique can avoid re -injury of posterior ligaments and bone structure, and may rebuild posterior tension band structure with the advantages of less trauma, less bleeding,shorter operative time, rapid postoperative recovery and so on.

Adult ; Bone Screws ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery

OBJECTIVETo study the mechanism of prophylactic effects of nasal tolerance with a dual analogue (Lys262-Ala207) on experimental autoimmune myasthenia gravis (EAMG).

METHODSClinical and immunological changes were observed in Lewis rats administered with dual analogue Lys262-Ala207 nasally, to compare the effects between the rats with predetermined dosage of Lys262-Ala207 and control peptides at two different time points, before the day (Group A or C) or on the day (Group B or D) of immunization with acetylcholine receptor (AChR) in complete Freud's adjuvant for 10 consecutive days. The clinical scores was evaluated for 50 days post immunization. Numbers of MNC expressing IFN-gamma, IL-4 or IL-10 and CD4+ and/or CD25+ from lymph nodes were enumerated by flow cytometry. Proliferative response, expressed as stimulation index (SI), was suppressed in response to antigen-specific stimulation in the rats receiving dual analogue, as compared with the rats receiving saline buffer only.

RESULTSGroup A and group B of Lewis rats developed EAMG with reduced severity, as compared to the control groups. Number of cells synthesizing IFN-gamma, IL-4 or IL-10 decreased, whereas numbers of CD4+CD25+ cells increased in group A and B than those in the control groups. Proliferative response was suppressed in response to antigen-specific stimulations in the rats receiving dual analogue Lys262-Ala207.

CONCLUSIONSNasal administration with a dual analogue Lys262-Ala207 at two different time points, before the day and on the day of immunization, could delay symptoms of muscular weakness in EAMG rats, which was associated with suppression of immune function in AChR antigen-specific T cells and lay a scientific foundation for treatment of human MG with nasal dual analogue.

Administration, Intranasal ; Animals ; Antibody Affinity ; Autoantibodies ; immunology ; Dose-Response Relationship, Immunologic ; Female ; Immune Tolerance ; drug effects ; Immunity, Cellular ; Immunity, Mucosal ; Lymphocyte Activation ; Myasthenia Gravis ; immunology ; prevention & control ; Nasal Mucosa ; immunology ; Rats ; Rats, Inbred Lew

OBJECTIVETo observe the protective effects of histone deacetylase inhibitor on stress-induced myocardial injury.

METHODSHealthy male Wistar rats were randomly divided into 3 groups( n = 6), and the stress-induced myocardial injury model was established with chronic restraint stress method. The protective effects of histone deacetylase inhibitor on stress-induced myocardial injury were observed with Trichostatin A (TSA) intervention. Histone acetylation levels in myocardium of rats were detected by Western blot method, spectrophotometry method was used to dynamically determine the activity of rat serum lactate dehydrogenase (LDH), serum creatine kinase isoenzyme-MB (CK-MB) and Caspase 3, and nagar Olsen staining were used to observe the early myocardial damage.

RESULTSRestraint stress could significantly reduce the level of histone acetylation of myocardium in rats, and TSA intervention could inhibit the stress-induced reduction of myocardial levels of histone acetylation. Restraint stress could cause the significant increase of serum LDH activity ( P < 0.05), serum CK-MB activity ( P < 0.05), and the Caspase 3 activity of myocardial tissue (P < 0.05), and early myocardial damage also occurred during restraint stress. ISA intervention could significantly reduce the serum LDH activity (P < 0.05), the serum CK-MB activity (P < 0.05), the activity of myocardial tissue caspase 3 induced by restraint stress (P < 0.05), and the stress-induced myocardial injury was also attenuated by TSA intervention.

CONCLUSIONThe histone deacetylase inhibitor TSA can protect stress-induced myocardial injury.

Acetylation ; Animals ; Cardiotonic Agents ; pharmacology ; Caspase 3 ; blood ; Creatine Kinase, MB Form ; blood ; Histone Deacetylase Inhibitors ; pharmacology ; Hydroxamic Acids ; pharmacology ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Restraint, Physical ; Stress, Physiological