Objective To compare the sensitivity and specificity of different fluorescent quantitation polymerase chain reaction(PCR) for genes detection of Streptococcus pneumonia.Methods By designing five sets of fluorescent quantitation PCR primers targeting different genes of Streptococcus pneumonia (lytA,ply,spn9802,psaA,cspA),the sensitivity and specificity of fluorescent quantitation PCR was determined through detecting 37 strains of Streptococcus pneumonia and 28 strains of non-Streptococcus pneumonia,as well as 80 sputum specimens from suspected pneumonia cases.Results These five primers had excellent sensitivity with 90％ amplification efficiency through analysis of the standard curve.The change of Ct value was less than 0.5,indicating that these five primers have good stability.By detecting 37 strains of Streptococcus pneumonia and 28 strains of non-Streptococcus pneumonia,it showed that the specificity of lytA,ply and spn9802 was better than the other two primers (positive rate and negative rate was all 100％).But the detection limit of lytA,ply was one order of magnitude than spn9802 (101 CFU/ml vs.102 CFU/ml).The specificity of psaA and cspA was worse [negative rate of psaA was 89％ (25/28),positive rate of cspA was 97％ (36/37)].By detecting 80 sputum specimens from suspected pneumonia cases,it showed that the specificity of fluorescent quantitation PCR were better than the bacterial culture.The specificity of lytA and ply was the best two [positive rate:92.50％(74/80),90.00％(72/80)],and next was spn9802 [86.25％(69/80)],the specificity ofpsaA and cspA was the worst two for detection [76.25％(61/80) and 78.75％(63/80)].Conclusions The sensitivity and specificity of fluorescent quantitation PCR genes used in clinic for detecting Streptococcus pneumonia is different.LytA and ply are the best primers in specificity,next is spn9802,and psaA and cspA are the worst primers for in specificity detection.

Objective To explore the value of three-dimensional constructive inference in steady state sequence (3D-CISS se-quence)in case of midbrain aqueduct obstruction.Methods 46 cases with midbrain aqueduct obstruction were scanned with FLASH T1 WI、TSE T2 WI and 3D-CISS sequence at 3.0T superconductive MR scanner.The original images of the 3D-CISS sequence were reconstructed.The images of three sequences showing midbrain aqueduct obstruction were observed and analyzed.Results The de-tection rate of the midbrain aqueduct obstruction was 13% 、71.7% and 100% in FLASH T1 WI、TSE T2 WI and 3D-CISS sequence, respectively.The difference between the three sequences were significant (P <0.01 67).Conclusion 3D-CISS sequence shows mid-brain aqueduct obstruction more accurately.

Critical value report procedures need to be improved rationally by urban space saving, post setting, and monitoring and analysis of historical data of critical value.Laboratory is required to develop and put into practice critical value policies and procedures by numerous regulatory agency standards, such as, International Standard Organization ( ISO 15189) , College of American Pathologists.Laboratories should implement and continuously improve quality management system for achieving the quality goals set by Laboratory and International Standard Organization.

Objective To evaluate potential clinical risk factors for the development of early-onset myocardial damage following multiple trauma (MT), and to determine whether early-onset myocardial damage was caused by the combined effects of thoracic and systemic injury factors in MT patients.Methods A total of 231 patients with MT over the last 3 years were retrospectively reviewed. With myocardial damage being a dependent variable and other twenty factors being independent variables, univariate and multivariate logistic regressions were applied to investigate the risk factors for early-onset myocardial damage and to identify the association of thoracic and systemic risk factors with early-onset myocardial damage.Results Multivariable logistic regressions showed that acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ ) score ≥10, injury severity score (ISS) ≥ 25, shock index ≥ 2,coexisting chest trauma, abbreviated injury scale (AIS) of chest≥3, and hypoxia time ≥ 0.5 h were risk factors. The risk of earlyonset myocardial damage following MT obviously increased when thoracic and systemic injury risk factors were coexisting.Conclusion Our results indicated that thoracic injury combined with systemic injury increased the overall risk of early-onset myocardial damage following MT. Prospective validation of these findings in other clinical settings is warranted.

Objective To observe the feasibility of PC-cine MRI for estimating the cerebrospinal fluid circulation of fistula after endoscopic third ventriculostomy (ETV)in obstructive hydrocephalus.Methods 25 cases with obstructive hydrocephalus were scanned with routine MR protocol and PC-cine sequence before and after ETV.Qualitative and quantitative evaluation of the cerebro-spinal fluid flow through the fistula were performed and the results were compared with 25 cases of healthy volunteers.Results One week after operation,the cerebrospinal fluid flow through the fistula showed obviously positive in 23 patients,while negative in the other 2 patients,and the waveform was similar to that in the normal midbrain aqueduct.The outflow and inflow peak velocity of the cerebrospinal fluid through the fistula were lower than that in normal midbrain aqueduct(P<0.05),while the quantity of outflow,inflow and netflow were much higher(P<0.05).6 patients showed ventricular narrowing one week after operation and 17 cases maintained the same finding.During follow-up of half year,the ventricular size still showed no obvious narrowing in 14 patients.Ventricular expanding was observed in 2 post-operative patients.Conclusion PC-cine MRI can provide intuitive and reliable evidences in evaluation of ETV for obstructive hydrocephalus.

OBJECTIVETo investigate the effect of kirenol on bovine type II collagen (CII)-specific lymphocytes in vivo and in vitro, and explore the mechanism of kirenol-induced immunosuppression in antigen-specific lymphocytes.

METHODSTwenty-four Wistar rats were randomized into control group, collagen-induced arthritis (CIA) model group, kirenol group (2 mg/kg), and prednisolone group (2 mg/kg). After CII injection, the rats in the latter two groups received intragastric administration of kirenol and prednisolone for 30 days, and the spleens and draining lymph nodes of the rats were harvested to prepare single cell suspensions for measurement of the cytokine levels using ELISA. In the in vitro experiment, the lymphocytes from the control rats, with or without 20 µg/ml CII treatment in the presence of 0-80 µg/ml kirenol, were evaluated for cell proliferation and apoptosis using [(3)H]-thymidine incorporation and flow cytometry, respectively.

RESULTSCompared with those in CIA group, IFN-γ and TNF-α production was significantly reduced in splenocyte culture supernatant of kirenol group (P<0.05 and P<0.01, respectively), and the level of IL-10 and IL-4 was up-regulated (P<0.05 and P<0.01, respectively); IFN-γ and TNF-α secretion by the cultured lymph node cells (LNCs) significantly decreased (P<0.05 and P<0.001, respectively) and IL-10 and IL-4 production increased (P<0.05, P<0.001) in kirenol group. In the in vitro experiment, kirenol treatment caused obvious suppression of CII-induced LNC proliferation and dose-dependently induced antigen-specific apoptosis of the splenocytes and LNCs.

CONCLUSIONKirenol treatment reduces pro-inflammatory cytokine secretion, increases anti-inflammatory cytokine production, inhibits cell proliferation and induces apoptosis of CII-specific lymphocytes in vitro, suggesting the potential of kirenol as an immunosuppressant.

Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Arthritis, Rheumatoid ; chemically induced ; drug therapy ; immunology ; Cattle ; Cell Proliferation ; Cells, Cultured ; Collagen Type II ; immunology ; Cytokines ; immunology ; Diterpenes ; pharmacology ; therapeutic use ; Female ; Immunosuppressive Agents ; pharmacology ; therapeutic use ; Lymphocytes ; cytology ; drug effects ; immunology ; Rats ; Rats, Wistar

Objective To investigate the significance of blood oxygenation level dependent functional magnetic resonance imaging (BOLD-fMRI) and ambulatory urodynamics monitoring (AUM) to find the cause of primary monosymptomatic nocturnal enuresis (PMNE) in children.Methods Thirty-seven children with PMNE (20 males and 17 females) were selected,with a mean age of (11.3 ±4.1) years old.A clinical management tool,3-day urination record,daytime BOLD-fMRI scan,conventional urodynamics (CUD) and AUM were performed respectively.Thirty-seven gender-age matched children(19 males,18 females) who would receive surgery treatment because of upper urinary tract disease and were confirmed to have no lower urinary tract dysfunction by CUD were enrolled as controls,with a mean age of (11.1 ± 2.9) years old,and 13 cases underwent BOLD-fMRI scanning.Results It was found that the ALFF value of the left middle frontal gyrus of PMNE was decreased and the ReHo value of the left superior occipital gyrus was increased compared with the control group by the resting BOLD-fMRI.The maximum voiding volume of PNME children was (303.11 ± 87.48) mL,the total urine volume at night was (568.65 ± 208.48) mL,and the nighttime bladder volume was (217.43 ± 81.53) mL.The incidence of maximum voiding volume reduction,nocturnal polyuria and decreased nocturnal bladder volume were 24.32％ and 56.76％,and 64.86％,respectively.However,AUM results showed that maximum detrusor pressure in the PMNE group was (39.22 ± 7.78) cmH2O (1 cmH2O =0.098 kPa),which was statistically significantly higher than that in CUD (32.22 ±9.00) cmH2O,and the difference was statistically significant (P ＜0.05).In PMNE group,29 cases (78.37％) had detrusor overactivity (DO),which was significantly higher than that in CUD group [16 cases (43.24％)],and the difference was statistically significant(t =-3.047,P =0.004).CUD and AUM were all detected in children with DO,the frequency of DO detected by AUM was significantly higher than that detected by CUD[(2.00 ±0.55 times/h) vs.(1.38 ±0.50) times/h,P ＜0.05],and the maximum amplitude of detrusor pressure when DO occurring was significantly higher in AUM than in CUD [(19.56 ± 6.01) cmH2O vs.(14.38 ± 3.07) cmH2O],and the differences were statistically significant (all P ＜ 0.05);however,there was no significant difference in bladder compliance detected by AUM or CUD (P ＞ 0.05).Conclusions The abnormal functions of the left middle frontal gyrus and left superior occipital gyrus,nighttime DO and the decrease of bladder capacity at night are considered to be key causes of enuresis in children with PMNE.AUM and resting state BOLD-fMRI evaluations are helpful in differentiating the pathogenesis of PMNE.

Objective: This study was carried out to analyze the clinical characteristics of pseudohypopara-thyroidism(PHP) type 1b, and to improve the understanding and diagnosis of the disease. Methods: Five patients with molecular diagnosis of pseudohypoparathyroidism type 1b in our hospital during 2018 were enrolled, their clinical data, biochemical indicators, imaging, and gene detection results were analyzed. Results: There were 4 females and 1 male, with low calcium, high phosphorus and high PTH serum concentrations. The onset age span was large and the onset symptoms were different. Family history may not be obvious. There was abnormal methylation of GNAS gene or deletion of exon STX16 in methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) detection while gene sequencing result was negative. Conclusion MS-MLPA detection was still needed in patients with suspected PHP but negative gene sequencing result. Different methylation abnormalities and copy number variations might be correlated with the onset symptoms and familial of type 1b PHP.

This study reported a family of mitochondrial diabetes mellitus complicated with chronic hereditary pancreatitis. A 18-year-old woman presented with self-reported hyperglycemia and chronic epigastric pain was admitted to our hospital. Clinical data and family history were collected. Mitochondrial gene sequencing and whole exon gene sequencing showed that the proband carried mutation of mt.3243A>G and heterozygous mutation of SPINK1 c. 194+ 2T>C, which was considered as mitochondrial diabetes mellitus with chronic pancreatitis.

ObjectiveTo investigate the role and possible mechanism of action of rhubarb decoction （RD） retention enema in improving inflammatory damage of brain tissue in a rat model of mild hepatic encephalopathy （MHE）. MethodsA total of 60 male Sprague-Dawley rats were divided into blank group （CON group with 6 rats） and chronic liver cirrhosis modeling group with 54 rats using the complete randomization method. After 12 weeks， 40 rats with successful modeling which were confirmed to meet the requirements for MHE model by the Morris water maze test were randomly divided into model group （MOD group）， lactulose group （LT group）， low-dose RD group （RD1 group）， middle-dose RD group （RD2 group）， and high-dose RD group （RD3 group）， with 8 rats in each group. The rats in the CON group and the MOD group were given retention enema with 2 mL of normal saline once a day； the rats in the LT group were given retention enema with 2 mL of lactulose at a dose of 22.5% once a day； the rats in the RD1， RD2， and RD3 groups were given retention enema with 2 mL RD at a dose of 2.5， 5.0， and 7.5 g/kg， respectively， once a day. After 10 days of treatment， the Morris water maze test was performed to analyze the spatial learning and memory abilities of rats. The rats were analyzed from the following aspects： behavioral status； the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） and the level of blood ammonia； pathological changes of liver tissue and brain tissue； the mRNA and protein expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （AKT）， and mammalian target of rapamycin （mTOR） in brain tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the MOD group， the RD1， RD2， and RD3 groups had a significantly shorter escape latency （all P<0.01）， significant reductions in the levels of ALT， AST， IL-1β， IL-6， TNF-α， and blood ammonia （all P<0.05）， significant alleviation of the degeneration， necrosis， and inflammation of hepatocytes and brain cells， and significant reductions in the mRNA and protein expression levels of PI3K， AKT， and mTOR in brain tissue （all P<0.05）， and the RD3 group had a better treatment outcome than the RD1 and RD2 groups. ConclusionRetention enema with RD can improve cognitive function and inflammatory damage of brain tissue in MHE rats， possibly by regulating the PI3K/AKT/mTOR signaling pathway.