Objective Exploring the relationship among plasma tumor necrosis factor alpha(TNF-?),endothelin-1(ET-1),interleukin-6(IL-6) and cardiovascular diseases in patients with sleep apnea.Methods ELISA method were used to measure circulating levels of TNF-?,ET-1 and IL-6 in 15 with SAS complicated with cardiovascular diseases and 15 normal subjects, matched in the age ,sex ,height and weight.Results Plasma TNF-?,ET-1 and IL-6 level had increased significantly in SAS group,as compared with normal subjects(P

Objective To observe the results of crown lengthening surgery on the teeth with subgingival defect.Methods Crown lengthening surgery was performed on 54 terior teeth,which were divided into two groups based on probing depth of pre-operation:group A (<3 mm) and group B (3-4 mm).All teeth were restored 8 weeks after operation and followed-up to 1 year.The parameters of plaque index (PLI),suleus bleeding index (SBI),maximal defect probing depth (PD) and mobility degree (MD) at different times were measured,respectively.Results PLI,BI and PD were significantly improved during the follow-up period (P<0.01).The success rates of both groups were 96.8% and 69.6%, respectively (P<0.01).No significant difference about MD in the group A one year after restoration (P>0.05),but a significant improvement in the group B (P<0.01).Conclusion Crown lengthening surgery is a good method on the teeth with subgingival defect,but one must select the right indication.

Objective To investigate the role of Akt signal pathway on the expression of monocyte chemoattractant protein-1 ( MCP-1 ) and nuclear transcription factor-κB (NF-κB) in renal tubular epithelial cells (HK-2) stimulated by albumin and to explore the mechanisms of action. Method The HK-2 cells were incubated in the presence of albumin (5,15,30 mg/mL) with or without Ly294002 (an inhibitor of Akt). Expression of mRNA was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).Expression of Akt and protein MCP-1 were assessed by Western blot. Electrophoretic mobility shift assay (EMSA) was used to detect the activation of NF-κB. q-test was used to evaluate the differences in means between groups. Results Compared with control group, the expression of MCP-1 mRNA remarkly increased. [Control group: 0.233 ±0.01; BSA(5 mg/mL) group: 0.285 ±0.04; BSA( 15 mg/mL) group:0.387 ± 0.02; BSA ( 30 mg/mL) group: 0.473 ± 0.05; BSA ( 30 mg/mL) + Ly294002 group: 0. 325 ±0.05, P ＜ 0.05 ]. The expression of MCP-1 protein in renal interstitum of operation group were remarkly increased too. [ Control group: 100 ± 15.1; BSA ( 5 mg/mL) group: 148 ± 19.3; BSA ( 15 mg/mL) group: 176±20.7; BSA(30 mg/mL) group: 263 ± 18.1; BSA(30 mg/mL) + Ly294002 group: 175 ± 18.0, P ＜0.05 ]. Albumin stimulated the expression of MCP-1mRNA and protein in a dose-dependent manner. Albumin remarkably increased the activity of NF-κB. Albumin enhanced the expression of Akt. Ly294002 inhibited albumin-induced the expression of NF-κB and partially decreased the level of MCP-1. Apositive correlation was noted between NF-κB activation and MCP-1 expression( r = 0.68 ,P ＜ 0.01 ). Conclusions Albumin-induces MCP-1 and NF-κB production via Akt signal pathway in renal tubular epithelial cells.

Objective To establish a new double-antibody sandwich ELISA to detect the antigen AP33(a 33 kDa adhesive protein) of Trichomonas vaginalis based on the quantum dots and magnetic beads.Methods After BALB/c mice were immunized by AP33,the multiclonal antibodies in the antiserum was conjugated with the quantum dots and magnetic beads by carbon diimine crosslinking method respectively.Then the antibodies combined with magnetic beads were coated to microwell of plate as capturing antibody,and the antibodies bound to quantum dots were regarded as the marked antibody which can be directly observed under fluorescence microscope and quatitated by spectrofluorometry.The specificity and sensitivity of our established system were investigated.Results AP33 was successfully detected at the concentration as low as 50 ng/ml by this with-filling method.No crossreaction was observed when this system was used to detect Trichomonas vaginalis and other common bacteria in the vagina.The accuracy was 88% and the specificity was 90%.Conclusion This new double-antibody sandwich ELISA to detect Trichomonas vaginalis is successfully prepared and of sound specificity and sensitivity.

Objective To explore the titre of serum anti-phospholipase A2 receptor antibodies (anti-PLA2 R antibodies) detected by the enzyme-linked immunosorbent assay (ELISA) to provide a more specific serological index for clinical diagnosis and disease judgment of membranous nephropathy (MN).Methods Thirty-four cases of MN confirmed by kidney biopsy ,32 inpatients with autoimmune diseases ,nephrotic syndrome and renal insufficiency in the nephrology department of our hospital and 12 persons un-dergoing physical examination were selected.The serum was collected for detecting anti-PLA2 R antibodies level.Then its diagnostic performance for diagnosing MM was analyzed by combining with serum TP ,ALB ,IgG ,IgA ,IgM ,C3 and C4 indicators.Results The medians of anti-PLA2 R antibodies titres in the MN group ,disease control group and healthy control group were 22.1 ,2.0 ,2.0 RU/mL respectively ,which had statistical difference between the MN group and other two groups (P<0.05).Seventeen cases of anti-PLA2R antibodies positive were in the MN group(positive rate50% )and 17 cases were negative,the disease control group and healthy control group all were negative.Its specificity and positive predictive value were 100% ,TP ,ALB and IgG had statistical difference between the MN group with the disease control group and healthy control group (P<0.05);the relative coefficients of anti-PLA2 R antibodies with TP ,ALB and IgG ,IgA ,IgM ,C3 and C4 were in turn -0.382 ,-0.344 ,-0.502 ,-0.295 ,0.062 , 0.005 and 0.241 respectively ,anti-PLA2R antibodies were negatively correlated with TP ,ALB ,IgG and IgA(P<0.01) ,positively correlated with C4(P<0.05) and had no relation with IgM and C3(P>0.05).Conclusion Anti-PLA2 R antibodies have higher specificity for the diagnosis of MN and can serve as the necessary and specific serologic detection indicator in the patients unable to conduct renal biopsy.Quantitative detection helps to condition judgment.

Objective To establish the quality standard of Zhilou Lotion. Methods TLC was adopted to identify Radix et Rhizoma Rhei and Cortex Phellodendri. HPLC was adopted to determine the content of chlorogenic acid in Zhilou Lotion. The chromatography column was Agilent Ecilipse XDB-C18 (4.6 mm× 250 mm, 5 μm), the mobile phase was acetonitrile-0.4% phosphoric acid solution (10∶90), the flow rate was 0.80 mL/min, the temperature of column was room temperature, and the detection wavelength was 327 nm. Results The method for identification had good specificity and repeatability. There was no interference in blank control. Chlorogenic acid had good linearity in the range of 2.442-122.1 μg (r=1.000 0). The average recovery rate was 100.30% and RSD was 1.05% (n=9). Conclusion The qualitative and quantitative method is simple, specific, accurate and reliable, and can be used for quality control of Zhilou Lotion.

AIM: To explore the effects of cellular signal transduction pathways of heme oxygenase-1(HO-1)-carbon monoxide (CO)-cyclic GMP (cGMP) and nitric oxide synthase (NOS)-nitric oxide (NO)-cGMP on cholecystokinin octapeptide (CCK-8) reversed vascular hyporeactivity in endotoxemic rats. METHODS: According to the treatments given in vivo , rats were devided into four groups: control; lipopolysaccharide (LPS); CCK and CCK+LPS. Using isolated vascular ring tension detecting technique, thoracic aortic rings (TARs) were prepared and accumulation of contractive responses to phenylephrine (PE) were measured under which the TARs were incubated with Hemin (He, donor of CO), Zinc-protoporphyrin-IX (ZnPP-IX, selective inhibitor of HO-1), L-arginine (L-Arg, substrate of NOS), aminoguanidine (AG, selective inhibitor of iNOS), N ?-nitro-L-arginine (L-NNA, inhibitor of NOS) or methylene blue (MB, inhibitor of guanylyl cyclase), respectively. RESULTS: CCK-8 alone did not affect vascular tension. Injection of LPS induced the hyporeactivity of the TARs and was reversed by pretreatment of CCK-8. In LPS and CCK+LPS groups, the hyporeactivity was partly reversed by incubation of TARs with ZnPP-IX or AG, and restored to normal by incubation of TARs with L-NNA or MB. Incubation of TARs with He or L-Arg showed to make the vascular hyporeactivity worse in different degree. CONCLUSIONS: CCK-8 alone did not affect the activity of HO-1 and iNOS but influenced the activity of these enzymes induced by LPS, which lead to reduced CO/NO production, decreased the content of cGMP and plays its important role in reversing vascular hyporeactivity in endotoxemic rats.

AIM: To verify the effect of cholecystokinin octapeptide(CCK-8) on cardiac function in endotoxin shock (ES) rats.METHODS: The rats were divided into four groups:control,lipopolysaccharide(LPS),CCK-8 and CCK-8+LPS.The left ventricle pressure(LVP),the maximal/minimum rate of LVP(?LVd p /d t max ),heart rate (HR) and mean arterial pressure (MAP) were measured.The activity of superoxide dismutase (SOD),the contents of malondialdehyde (MDA) and nitric oxide (NO) in both serum and myocardium were also measured,respectively.RESULTS: CCK-8 (40 ?g?kg -1 , iv) elicited bradycardia in short time and gently increase MAP,LVP and ?LV d p /d t max . Lipopolysaccharide(LPS, 8 mg?kg -1 , iv) caused a variation in heart rate (HR)(a bradycardia following a tachycardia) and rapid decreases in MAP,LVP and ?LVd p /d t max . The rapid variation of HR and the decline of MAP,LVP and ?LVd p /d t max were reversed by pretreatment with CCK-8 in ES rats, but didn't restore to normal.The activity of SOD was increased and the contents of MDA and NO were decreased by pretreatment with CCK-8 in ES rats.CONCLUSION: The decline of cardiac function in ES rats could be reversed by pre-administration of CCK-8 and the decrease in NO production may be one of the mechanisms.

BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.

Objective To investigate the rehabilitative effect of low frequency repetitive transcranial magnetic stimulation (rTMS) on convalescing patients with Broca's aphasia. MethodsTwenty-eight patients with Broca's aphasia recovering from cerebral infarction were randomly divided into a stimulation group and a control group with 14 subjects in each.Patients in the control group accepted conventional drugs,speech rehabilitation and sham stimulation,while patients in the stimulation group were in given low frequency rTMS in place of the sham stimulation.Their speech performance was evaluated using the China Rehabilitation Research Center's aphasia examination (CRRCAE) pre-stimulation,post-stimulation and 90 days later. ResultsCompared with before treatment and with the controls,the speaking scores of the stimulation group increased significantly after treatment and also 90 days later. ConclusionLow frequency rTMS can not only improve the speech performance of Broca's aphasia sufferers in the short term,but it also plays a lasting role.It may thus have clinical application for patients with Broca's aphasia.