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Meningoencephalitis caused by the human herpes virus 6 is common in acquired immune deficiency syndrome patients or in patients after organ transplantation, rarely in immunocompetent adults. The clinical manifestations include headache, drowsinesss, coma, epileptic attack, psychiatric symptom and focal neurological signs. Antiviral therapy is effective, however the prognosis should be poor if the treatment is not timely. The clinical data of one case of meningoencephalitis caused by the human herpes virus 6 in Tianjin Haihe Hospital in October 2019 were analyzed. Meningoencephalitis caused by the human herpes virus 6 can be recurrent. The clinical manifestations are nonspecific and the diagnosis is difficult. The next generation sequencing of cerebrospinal fluid is a powerful method to identify pathogens.
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OBJECTIVE@#To develop a cell-based system for the diagnosis of vitamin K-dependent coagulation factor deficiency 1 (VKCFD1).@*METHODS@#In HEK293 cells stably expressing the reporter gene FIX-Gla-PC, the gamma-glutamyl carboxylase (GGCX) gene was knocked out by using CRISPR/Cas9 technology. Enzyme-linked immunosorbent assay (ELISA), DNA sequencing and Western blotting were used to identify the GGCX gene knockout cells. A quickchange point variant method was used to construct the GGCX variant. ELISA was used to assess the influence of GGCX variant on the activity of reporter gene.@*RESULTS@#Two monoclonal cell lines with no reporter activity by ELISA was identified. Edition and knockout of the GGCX gene was confirmed by DNA sequencing and Western blotting. The activity of the reporter gene was recovered by transfection of the wild-type GGCX gene. Thereby two monoclonal cells with GGCX knockout were obtained. By comparing the wild-type and pathogenic GGCX variants, the reporter activity was decreased in the pathogenic variants significantly.@*CONCLUSION@#A cell-based system for the detection of GGCX activity was successfully developed, which can be used for the diagnosis of VKCFD1 caused by GGCX variants.
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Objective:To observe the expression, correlation and significance of chemokine (C-X-C motif) ligand 12 (CXCL12) and chemokine (C-X-C motif) receptor 4 (CXCR4) in endometrium and myometrium of adenomyosis.Methods:Totally 38 patients were selected in this study, who underwent hysterectomy for adenomyosis at Beijing Obstetrics and Gynecology Hospital from October 2017 to December 2018 as the adenomyosis group, and, in the same period, selected 31 patients with cervical intraepithelial neoplasia Ⅲ or cervical cancer undergoing hysterectomy served as control group. The expression levels of mRNA and protein for CXCL12, CXCR4 in the endometrium and myometrium of the two groups were detected by immunohistochemistry and real-time PCR.Results:(1) The protein levels of CXCL12 and CXCR4 in endometrium in uterus with adenomyosis (0.229±0.025 and 0.226±0.016) were significantly higher than those in endometrium in uterus without adenomyosis (0.153±0.018 and 0.178±0.026); compared with each other, the differences were statistically significant (all P<0.05). And the expressions of CXCL12 and CXCR4 proteins in uterine myometrium of adenomyosis were 0.222±0.045 and 0.126±0.058, respectively, which were higher than those in the control group (0.091±0.029 and 0.099±0.020); compared with each other, the differences were statistically significant (all P<0.05). (2) The expression levels of CXCL12 and CXCR4 mRNA in endometrium of patients with adenomyosis were 6.31±0.12 and 8.49±0.21, respectively, which were higher than those in the control group (1.23±0.10 and 1.36±0.13); compared with each other, the differences were statistically significant (all P<0.05). Moreover, the expression levels of CXCL12 and CXCR4 mRNA in myometrium of patients with adenomyosis were 9.11±0.12 and 8.45±0.16, respectively, which were higher than those in the control group (1.18±0.08 and 1.46±0.13); compared with each other, the differences were statistically significant (all P<0.05). (3) In endometrium and myometrium of uterus with adenomyosis, CXCL12 and CXCR4 mRNA expression levels were positively associated ( r=0.478, 0.542, all P<0.05). Conclusions:The levels of CXCL12 and CXCR4 in the endometrium and myometrium of adenomyosis are increased and positively correlated. The two chemokine may be involved in the development of adenomyosis.
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Objective:To investigate the serum complement C1q level in patients with polymyositis/dermatomyositis (PM/DM) and its clinical significance.Methods:Thirty patients with PM/DM admitted to the Department of Endocrinology in Shuguang Hospital from January 2016 to December 2018 were selected as the case group. Thirty healthy people for physical examination in the same period in Shuguang Hospital were selected as the control group. The levels of complement C1q and creatine kinase (CK) in the serum of all the subjects were detected. Meanwhile, the levels of anti-Jo-1 antibody(Jo-1) , antineutrophil cytoplasmic antibodies (ANCA) and anti-ribonuclear protien antibody (RNP) in PM/DM patients were detected. The levels of complement C1q and CK in the serum of PM/DM patients with different clinical symptoms and antibody expression were compared. The working characteristics (ROC curve) were used to determine the diagnostic efficacy of complement C1q and CK for PM/DM.Results:Compared with the control group, the levels of complement C1q [(241.26±26.10) mg/L vs (125.04±11.87)mg/L] and CK[(561.03±29.48)mU/ml vs (161.74±15.86)mU/ml] in serum of patients with PM and DM were higher ( t=22.201, P=0.026; t=65.331, P=0.012) , but there was no significant difference in the levels of complement C1q [(254.63±29.24) mg/L vs (240.18±25.72) mg/L] and CK[(557.19±50.10)mU/ml vs (585.42±63.64)mU/ml] between PM and DM( t=1.427, P=0.165; t=1.358, P=0.185). The serum complement C1q and CK in patients with myalgia, limb muscle weakness, Reynolds phenomenon, arthralgia, Yang rash, interstitial lung disease, Gottron sign, V sign, dysphagia and other organ-accumulated patients were increased( P<0.05). The levels of complement C1q[(189.25±24.20) mg/L vs (352.19±30.22) mg/L; (207.31±16.29)mg/L vs (298.16±34.29) mg/L] and CK[(524.56±51.32) mU/ml vs (752.56±49.61)mU/ml; (497.36±54.28) mU/ml vs (651.22±58.26)mU/ml] in patients with serum of Jo-1 or ANCA positive were lower than those of negative patients (χ 2=8.536, P=0.019;χ 2=7.694, P=0.037 and χ 2=12.696, P=0.027; χ 2=10.763, P=0.046). The levels of complement C1q [(316.00±34.82) mg/L vs (225.46±41.38) mg/L] and CK[(624.18±26.10)mU/ml vs (468.25±24.27)mU/ml] in serum of RNP positive patients were higher than those of negative patients (χ 2=6.712, P=0.041; χ 2=11.751, P=0.038). The ROC curves of complement C1q and CK in diagnosis of PM/DM were 0.842 and 0.783 respectively, and the Jordan index was 0.811 and 0.534 respectively, which had certain value for diagnosis of PM/DM, but that of the complement C1q level was higher than that of CK ( P<0.05). The sensitivity and specificity of complement C1q were 88.95% and 92.10% respectively with 231.820 mg/L as the critical value, which was higher than that of CK (68.42% and 85.00%)( P<0.05). Conclusions:The expression of complement C1q in serum of PM/DM patients is increased, which is related to the clinical symptoms and antibody levels of PM/DM patients. It may be involved in the progression of PM/DM disease.
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Objective To investigate the advantages and continuous optimization of laboratory automation system through analysis and assessment of the core data and performance after the application of open assembly line.Methods Collect the data of biochemical and immunoassay in Shuguang Hospital attached to Shanghai University of Traditional Chinese Medicine from April to October 2017.( 1 ) Cost analysis of the assembly line schemes;(2) Analysis of workflow before and after the application of assembly line;(3) Analysis of the volume of samples collecting before and after the application of assembly line ;(4) Analysis of TAT data before and after the application of assembly line; ( 5 ) Analysis of staffs allocation before and after the application of assembly line; (6) Analysis of samples rechecking before and after the application of assembly line .Results (1) Open assembly line costs least on hardware (8 million) and site among various projects;(2) Inspection process is greatly simplified after the application of assembly line;(3) The samples′volume of biochemical and immunoassay inspection were reduced by 31.85％;(4) The items′test cycle decreases after the application of assembly line , the average TAT is reduced by 32 minutes;(5) Staffs for samples pretreatment can be reduced by 50％after the application of assembly line , and the quantity of operators does not change;(6) The number of re-check samples increase except the gray zone and critical values , which ensures the reliability of the results .Conclusion To analyze the core data and to evaluate the performance , the laboratory improve on detection cycle ,staffs,and test efficiency.
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Objective To analyze the clinical application of detecting anti Mullerian hormone (AMH) in the patients with polycystic ovary syndrome (PCOS).Methods A total of 480 cases with PCOS and 500 cases of healthy females were selected in the study.The levels of AMH in the patients with PCOS were compared between the two groups.Results The levels of AMH,luteinizing hormone (LH),testosterone (T) in patients with PCOS were significantly higher than those in the healthy control group (P＜0.05).Differences of the levels of estradiol (E2) and follicle-stimulating hormone (FSH) in patients with PCOS and thosein healthy control group had no statistical significance(P＞0.05).As the age increased,the level of AM H obviously declined.Conclusion The level of AMH in patients with PCOS is obviously higher than that in the healthy ones,it declines with the age increasing.
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Objective To analyze the clinical features of the patients with meningitis caused by the Campylobacterfetus.Methods The clinical data of one case of meningitis caused by the Campylobacter fetus in Tianjin Haihe Hospital in September 2016 were analyzed and the literature was reviewed.In the PubMed database with “ Campylobacter fetus,meningitis” as keyword,we retrieved 51 English literature,from 1962 to 2016,excluding infant infection and non infection meningitis.Results Finally 17 articles,20 patients,were retrieved.With these patients,a total of 21 patients were analyzed.There were 19 male cases,two female cases,average age being 47 years (23-84 years).The clinical manifestations were fever,headache,some patients with altered consciousness,increased white blood cells and protein in cerebrospinal fluid,normal or decreased glucose.Antibiotic treatment was effective,and the prognosis was good.No specific clinical fetures were found.Alcoholism,diabetes and immunodeficiency were independent risk factors of the disease.Positive cerebrospinal fluid and blood cultures can help to diagnose the disease.Conclusions Campylobacter fetus caused meningitis in clinic is extremely rare,and its clinical manifestations are nonspecific.Antibiotic therapy is effective,and the prognosis is good.
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Ribosomal S6 kinase is a family of serine/threonine protein kinases involved in the regulation of cell viability. There are two subfamilies of ribosomal s6 kinase, (p90rsk, p70rsk). Especially, p90rsk is known to be an important downstream kinase of p44/42 MAPK. We investigated the role of p90rsk on ethanol-induced cell proliferation of HepG2 cells. HepG2 cells were treated with 10~50 mM of ethanol with or without ERK and p90rsk inhibitors. Cell viability was measured by MTT assay. The expression of pERK1, NHE1 was measured by Western blots. The phosphorylation of p90rsk was measured by ELISA kits. The expression of Bcl-2 was measured by qRT-PCR. When the cells were treated with 10~30 mM of ethanol for 24 hour, it showed significant increase in cell viability versus control group. Besides, 10~30 mM of ethanol induced increased expression of pERK1, p-p90rsk, NHE1 and Bcl-2. Moreover treatment of p90rsk inhibitor attenuated the ethanol-induced increase in cell viability and NHE1 and Bcl-2 expression. In summary, these results suggest that p90rsk, a downstream kinase of ERK, plays a stimulatory role on ethanol-induced hepatocellular carcinoma progression by activating anti-apoptotic factor Bcl-2 and NHE1 known to regulate cell survival.
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Humans , Blotting, Western , Carcinoma, Hepatocellular , Cell Proliferation , Cell Survival , Enzyme-Linked Immunosorbent Assay , Ethanol , Hep G2 Cells , Phosphorylation , Phosphotransferases , Protein Kinases , Ribosomal Protein S6 KinasesABSTRACT
Objective To investigate the correlation of serum levels of retinol binding protein 4 (RBP4) and andomen?tin-1 with ischemic stroke and carotid artery stenosis. Methods Healthy adult (n=50) were selected as the control group while ischemic stroke patients with different degrees of carotid artery stenosis (n=143) were enrolled as the disease group. According to the ultrasonic Doppler performance, these ischemic stroke patients were divided into three groups:mild steno?sis group (carotid artery stenosis was normal or less than 50%, n=67), moderate stenosis group (carotid artery stenosis was 50%to 70%, n=45), severe stenosis group (carotid artery stenosis was 70%to total occlusion, n=31). BMI index, hyperten?sion, total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL) and fasting blood glucose (FPG) were detected by Automatic biochemical analyzer and serum levels of RBP4 and omentin-1 were measured by ELISA. Results Hypertensive population in carotid artery stenosis group was larger than that in control group (P<0.05). Serum TC, TG, LDL, FPG and RBP4 levels were significantly higher while omentin-1 level decreased remarkably in disease group than those in the control group (P<0.05). Logistic regression analysis showed that TC and RBP4 were the risk factors of ischemic stroke with carotid artery stenosis, while omentin-1 was a protective factor. Conclusion RBP4 is a risk factor for ischemic stroke with carotid artery stenosis and omentin-1 is a protective factor.
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To see the inhibitory mechanism of gentamicin in response to electrical field stimulation (EFS) using the rat bladder smooth muscle, atropine or guanethidine was treated but had no effect. Methylsergide, a non-selective 5-HT1, 5-HT2 receptor antagonist was also treated but had on effect. Kinase inhibitors, such as chelerythrine (PKC inhibitor), ML-9 (MLCK inhibitor), or Y27632 (rho kinase inhibitor) were pretreated before gentamicin treatment, but did not have effect. For U73122, a phospholipase C (PLC) inhibitor however, the inhibitory effect to gentamicin was significantly attenuated in all frequencies given by the EFS. Therefore gentamicin induced inhibitory effect on EFS response in rat bladder smooth muscle was not mediated by the activation of adrenergic, cholinergic, or serotonergic receptor. The inhibition of gentamicin might be mediated through the PLC dependent pathway, but not through the PKC, MLCK or rho kinase dependent pathway.
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Animals , Rats , Atropine , Gentamicins , Guanethidine , Muscle, Smooth , Phosphotransferases , rho-Associated Kinases , Type C Phospholipases , Urinary BladderABSTRACT
A number of studies have demonstrated that 5-hydroxytryptamine (5-HT) can induce muscle contraction or relaxation response and enhance secretion in the gastrointestinal tract via a multiplicity of 5-HT receptor subtypes. In the present study, we investigated the pharmacological characterization of the 5-HT-induced contractile response in longitudinal smooth muscle isolated from the feline ileum. Addition of 5-HT into muscle chambers enhanced the basal tone and spontaneous activity in a concentration-dependent manner. The neurotoxin tetrodotoxin did not alter the 5-HT-induced contraction of the longitudinal muscles. Neither atropine nor guanethidine affected the contraction. The 5-HT agonists, 5-methylserotonin hydrochloride and mosapride, also evoked concentration-dependent contractions. The 5-HT-induced contraction was enhanced by the 5HT2 receptor antagonist ketanserin and the 5-HT3 receptor antagonist ondansetron but was inhibited by the 5-HT1 receptor antagonist methysergide and 5-HT4 receptor antagonist GR113808. These results indicate that 5-HT1 and 5-HT4 receptors may mediate the contraction of the 5-HT-induced response and 5-HT2 and 5-HT3 receptors may mediate 5-HT-induced relaxation in feline ileal longitudinal smooth muscles.