Objective To investigate the clinical application value of fluorescence PCR in vitro diagnosis(IVD)reagent kits in the HLA-B27 detection by comparing 2 kinds of HLA-B27 IVD reagent kit approved by CFDA.Methods A total of 573 clinical blood samples were collected and detected for HLA-B27 by the approved reagent kits based on the fluorescence PCR technique and the flow cytometry.The samples with inconsistent testing results by the two kits were further confirmed by the PCR sequencing.At the same time,about 5% samples of the positive results detected by the fluorescence PCR method were extracted for conducting the re-testing.Results Among 573 samples,191 samples were HLA-B27 positive and 382 cases were HLA-B27 negative by flow cytome-try;the same samples had 194 cases of HLA-B27 positive and 379 cases of HLA-B27 negative by real-time PCR.With flow cytome-try as reference of the final results,the positive coincidence rate of the two kinds of kit was 96.33%(184/191),the negative coinci-dence rate was 94.76%(362/382),27 samples had inconsistent results from the two kinds of assay(accounting for 4.71% of the to-tal number of samples),the total coincidence rate was 95.29% [(184+362)/573],the Kappa value was 0.896(P =0.02);the chi-square test P =0.021,the two kinds of testing method had the high consistency,but the differences existed in the testing results. The re-testing results by PCR sequencing(including 27 samples with inconsistent results by two kinds of kit)were entirely consist-ent with the fluorescence PCR testing results.Conclusion Compared with the authority method flow cytometry for HLA-B27 tes-ting in clinic,the fluorescence PCR kit may present more accurate judging ability for the HLA-B27 testing on the basis of ensuring the higher consistency of the testing results,is easier compared with the sample preparation and operating procedures,and has the stronger clinical application value and prospects s.

Objective To investigate the relationship of TCM constitution and syndrome elements based on patients with hyperhomocysteinemia. Methods A survey was conducted among 1316 patients with diabetes, coronary disease, hypertension and cerebral infarction. Totally 326 patients with hyperhomocysteinemia were incorporated in diagnostic data and their constitutions were determined. At the same time, syndrome elements identification method for syndrome elements was applied for judgment, and consistency of the constitution and syndrome elements was analyzed. Results The survey showed that prevalence rate of population with hyperhomocysteinemia was 24.77%;there were 6 cases with mild nature, and 320 cases with biased constitution;phlegm dampness, blood stasis, qi deficiency of constitutions 110 cases, 73 cases and 59 cases, respectively;blood stasis, phlegm, qi deficiency, and dampness were the main syndrome elements of hyperhomocysteinemia, of 100 cases, 98 cases, 73 cases and 59 cases, respectively;the consistency of phlegm dampness constitution and syndromes of phlegm was the most obvious (Kappa=0.89, P<0.05). Conclusion Phlegm dampness, blood stasis, and qi deficiency were the most common constitutions of hyperhomocysteinemia;blood stasis, phlegm, qi deficiency and dampness were the main syndrome elements of hyperhomocysteinemia patients. The two results were consistent, and the consistency of phlegm syndrome and phlegm dampness constitution was the most obvious.

Objective To investigate the changes in cathepsin B (CatB) expression in acutely photodamaged human dermal fibroblasts (HDFs) and their significance.Methods HDFs were isolated from the foreskin of children,and subjected to primary culture and subculture.The fourth-to eighth-passage HDFs were used in the following experiment.HDFs were divided into two groups to receive irradiation with different doses of ultraviolet A (UVA) for different durations (acutely photodamaged group) or remain unirradiated (control group).Cell counting kit-8 (CCK8) assay was conducted to evaluate the proliferative activity of HDFs after irradiation with UVA at 5,10,15,20 and 25 J/cm2 respectively.Western blot and quantitative real-time reverse transcription PCR were performed to measure the protein and mRNA expressions of CatB respectively in HDFs at 24,48 and 72 hours after exposure to UVA at 10 J/cm2,and at 48 hours after exposure to UVA at 10,15,20 and 25 J/cm2.Statistical analysis was carried out by analysis of variance and least significant difference (LSD) test using the SPSS 13.0 software.Results UVA radiation induced a decrease in the proliferative activity of HDFs.When the dose of UVA was ≤ 10 J/cm2,the survival rate of HDFs maintained higher than 85％,and significant differences were observed in cell survival rate between unirradiated and irradiated HDFs at 24,48 and 72 hours (all P ＜ 0.05).Western blot showed that the gray value of CatB protein in the acutely photodamaged group irradiated with 10 J/cm2 UVA was significantly higher than that in the control group at 24 hours (0.76 ± 0.14 vs.0.35 ± 0.01,P ＜ 0.05),48 hours (1.34 ± 0.38 vs.0.45 ± 0.12,P＜ 0.05) and 72 hours (0.82 ± 0.09 vs.0.61 ± 0.06,P＜ 0.05).Increased mRNA expressions of CatB were also observed in the acutely photodamaged group compared with the control group at 24 hours (0.149 ± 0.009 vs.0.089 ± 0.015,P ＜ 0.05),48 hous (0.173 ± 0.009 vs.0.091 ± 0.010,P ＜ 0.05) and 72 hours (0.185 ± 0.158 vs.0.111 ± 0.017,P ＜ 0.05) after UVA radiation at 10 J/cm2.The gray value of CatB protein was 0.99 ± 0.07,1.49 ± 0.14,1.89 ± 0.08,2.07 ± 0.06 in HDFs at 48 hours after exposure to UVA of 10,15,20 and 25 J/cm2,respectively,significantly higher than that in the control group (0.60 ± 0.05,all P ＜ 0.05).Similarly,the mRNA expression of CatB was up-regulated in HDFs at 48 hours after UVA radiation at 10,15,20 and 25 J/cm2 compared with the unirradiated HDFs.Conclusion The protein and mRNA expressions of CatB are up-regulated in acutely photodamaged HDFs induced by UVA radiation.

Adult ; Cryptococcosis ; Cryptococcus neoformans ; Humans ; Lung Diseases, Fungal ; Male

ObjectiveTo investigate the effects of chitosan/PVA nerve conduits which used for repairing sciatics nerve defect in rats.MethodsTwenty-seven rats were divided into 3 groups randomly,with 9 rats in each group. Firstly, the 15mm defects in the left sciatic nerves were made in the rats and were respectively repaired with chitosan/PVA conduits graft (group A), the silicon conduits graft (group B),and autografts (group C). At 12 weeks after the operations, the left sciatic nerves were taken out, and the comparative evaluation was made on the repairing effects by wet weight of gastrocnemius and soleus muscles, histological examination,computerized imaging analysis and True Blue retrograde tracing. ResultsThe wet weight of gastrocnemius and soleus muscles showed no significant difference between the chitosan/PVA graft and autograft groups (P ＞ 0.05). The wet weight of gastrocnemius and soleus muscles in significant difference between the chitosan/PVA graft and the silicon group at 12 weeks after the operation(P ＜ 0.05). The nerve fiber density showed no statistically significant differences between the chitosan/PVA and autograft groups(P＞ 0.05).The regenerative nerve fiber in group B had normal morphological and structural characters under transmission electron microscope.True Blue-labeled neuron cell bodies were found within both anterior horn of gray matter in the spinal cord and dorsal root ganglions (DRGs) ipsilateral to the operated side of the tested rats on illumination with ultra-violet light 1 week after the injection of True Blue.Conclusion Chitosan/PVA nerve conduit can effectively promote the nerve regeneration and myelinization of rat sciatic nerve, which is expected to substitute for autograft to repair nerve defects succesfully.

Objective To study the effects of the soil factors on the yield and quality of Angelica dahurica originated from Sichuan Province.Methods The yield and contents of imperatorin,isoimperatorin,and other 11 different inorganic elements of A. dahurica,gathered from 15 different habitats of Suining in Sichuan Province were determined.And the basic soil nutrients and other 11 different mineral elements of the soil were analyzed.The effects of the soil factors on the yield and quality were analyzed through correlation analysis.Results The weight,length,circumference,and the contents of imperatorin and isoimperator in the roots of A.dahurica from different habitats were different.And the basic soil nutrients and other 11 different mineral elements of soil were different.The positive correlation between root length of A.dahurica,and soil available P content,root circumference of A.dahurica,and soil organic matter content,root weight of A.dahurica,and soil available P and K contents were all significant.The soil total nitrogen content had negatively significant correlation with the content of isoimperatorin.The correlation among the imperatorin,isoimperator contents of A.dahurica,and 11 other soil inorganic elements was not significant.Conclusion The soil factors have effects on the yield and quality of A.dahurica to some degree.

AIM: To evaluate effects of Pinaverium Bromide on different segments and layers of colonic smooth muscle in wrap restraint stress (WRS) rats and explore its possible therapeutic mechanism on different types of irritable bowel syndrome (IBS). METHODS: Adult SD rats were randomly divided into model group (wrap restraint stress group) and control group. Colonic smooth muscle strips were made from different segments and layers in two groups. The spontaneous contraction activities of colonic longitudinal/circular muscle (LM/CM) strips of rats were observed with organ bath system before and after addition of series concentrations of pinaverium. RESULTS: Pinaverium Bromide caused concentration-dependent inhibition of colonic smooth muscle, the inhibitory effect of pinaverium in model group was significantly stronger than that in control group(proximal colon: 28.54±4.82 vs 7.48±1.65,21.75±1.00 vs 12.56±3.15; distal colon: 15.71±5.27 vs 3.89±1.16, 20.16±3.16 vs 7.56±1.96 )(P<0.05). Compared with that of distal colon, inhibitory effect of pinaverium was significantly higher of proximal colon (P<0.05). For the inhibition of pinaverium, there was no significant difference between LM and CM strips in the same intestinal segments (P>0.05). CONCLUSION: Effects of Pinaverium Bromide on different colonic muscle layers and segments in WRS rats is probably related with its therapeutic mechanism on different types of IBS.

ObjectiveTo explore the effects and possible mechanisms of VCR combined with low dose cyclophosphamide(CTX) intermittently to treat severe systemic lupus erythematosus(SLE).It is assumed that this might be a new combination therapy for SLE and expected to improve the overall prognosis and outcome of SLE.MethodsMurine chronic graft-versus-host disease(cGVHD) model were developed for study.They were randomly divided into the control group,vincristine (VCR) pulse therapy group,CTX pulse therapy group,CTX every other day(EOD) group,VCR+CTX combination group.One way ANOVA and repeated measure variance analysis were used for statistical analysis.Results① Six weeks after cGVHD models were set up,the average 24-hour urine protein quantification was(5.02±0.88) mg,anti-dsDNA antibody was positive,and Ⅳ LN pathology could be observed histologically in the model murine.So cGVHD models were successfully developed.② Significantly difference in decreasing of 24-hour urine protein quantification was found in the CTX EOD group,VCR+CTX combination group and other groups (P＜0.01).Significant decrease in Cr,ALT,anti-dsDNA,was found in the CTX EOD group,VCR+CTX combination group,CTX pulse therapy group and other groups(P＜0.05).Decrease in urine MCP-1 and TGF-β1 could be detected,and statistical significant difference in these parameters could be found in the CTX EOD group,CTX pulse therapy group,VCR+CTX combination group and other groups (P＜0.01).MCP-1 and TGF-β1'expression in model kidney were reduced in the CTX EOD group,VCR+CTX combination group and had statistical significant difference in the CTX EOD group,VCR+CTX combination group,VCR pulse therapy group,and CTX pulse therapy group.③ VCRand CTX combination treatment was effectivein 24-hour urine protein quantification,blood Cr,ALT,anti-dsDNA and urine MCP-1,as well as urine TGF-β1 (P＜0.05).Conclusion ① The combination of VCR and CTX is synergistic in decreasing 24-hour urine protein quantification,Cr,and the expression of MCP-1,TGF-β1.② The adverse effect of VCR+CTX combination group is similar to VCR pulse therapy group and CTX pulse therapy group.

BACKGROUND:Animal experiments and clinical studies have shown that, functional deprivation at the early development stage may possibly cause nerve and cognition dysfunction at adulthood. However, no research focuses on the effect of olfactory function on the neurogenesis in piriform cortex of adult guinea pigs.
OBJECTIVE:To establish olfactory deprivation models and explore the effect of olfactory experience on the distribution of doublecortin, parvalbumin, glutamic acid decarboxylase 67 and the co-localization of doublecortin with neuron-specific nuclear-binding protein in piriform cortex of adult guinea pigs.
METHODS:Twenty guinea pigs were randomly divided into two groups. Olfactory deprivation models were established through unilateral naris-occlusion and the occluded animals were al owed to survive 3 and 6 weeks, then the specimens were harvested. The distribution of doublecortin, parvalbumin, glutamic acid decarboxylase 67 was detected with immunohistochemistry. The co-localization of doublecortin with neuron-specific nuclear-binding protein in piriform cortex of adult guinea pigs was determined with immunofluorescence method.
RESULTS AND CONCLUSION:The number of doublecortin, parvalbumin, glutamic acid decarboxylase 67 positive cel s in piriform cortex at the undeprived side was significantly higher than that at the deprived side at 3 and 6 weeks (P<0.05). There was no significant difference in the co-localization of doublecortin with neuron-specific nuclear-binding protein in piriform cortex at 3 weeks group and 6 weeks group (P>0.05). The findings suggest that olfactory experience promotes the maturation and differentiation of neurons in piriform cortex of adult guinea pigs.

Objective To investigate the role of Toll like receptor 4/nuclear factor-κB (TLR4/NF-κB) signal pathway on myocardial dysfunction after cardiac arrest-cardiopulmonary resuscitation (CA-CPR) in animal model. Methods Twenty-six pigs were randomly divided into sham group (n = 6), CA-CPR 12 hours group (n = 10) and CA-CPR 24 hours group (n = 10). The model of CA-CPR was reproduced by endocardial electrical stimulation for 8 minutes followed by CPR, and the pigs in sham group were only given anesthesia and tracheal intubation. The changes in hemodynamics including mean arterial pressure (MAP) and cardiac output (CO), as well as morphology and ultrastructure of myocardial cells were observed before and after CPR. The levels of serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined by enzyme linked immunosorbent assay (ELISA), and protein and mRNA expressions of TLR4/NF-κB in the myocardium were determined by Western Blot and real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR), respectively. Results Hemodynamic disturbance and myocardial serious injury were observed in CA-CPR groups. Compared with sham group, the levels of serum TNF-α were markedly increased 0.5 hour after return of spontaneous circulation (ROSC) in CA-CPR 12 hours and 24 hours groups (pg/L: 62.49±6.66, 48.39±2.37 vs. 10.75±0.74, both P < 0.05), and peaked at 2 hours (pg/L: 70.93±5.51, 66.03±2.60 vs. 10.87±0.91, both P < 0.05) followed by a gradual decline. The levels of serum IL-6 at 0.5 hours after ROSC in CA-CPR 12 hours and 24 hours groups were markedly higher than those of sham group (pg/L: 14.42±1.99, 11.23±1.12 vs. 8.75±0.74, both P < 0.05), and peaked at 12 hours (pg/L: 36.50±2.91, 38.15±1.26 vs. 8.88±0.62, both P < 0.05) followed by a gradual decline. The protein expressions of TLR4 and NF-κB in the myocardium were significantly increased in CA-CPR 12 hours and 24 hours groups as compared with sham group [TLR4 protein (gray value): 0.11±0.03, 0.24±0.05 vs. 0.05±0.02; NF-κB protein (gray value): 0.27±0.04, 0.24±0.03 vs. 0.09±0.02, all P < 0.05]. The mRNA levels of TLR4 in CA-CPR 12 hours and 24 hours groups were increased by approximately (9.93±1.07) folds and (9.21±1.27) folds of sham group respectively, and NF-κB mRNA expressions were increased by (4.44±0.96) folds and (6.09±0.81) folds of sham group respectively (all P < 0.01). Conclusion Activation of TLR4/NF-κB signal pathway may be one of the main pathological mechanisms of post resuscitation myocardial injury in a porcine model of CA-CPR.