Objective To assess the curative effect of microscopic anterior cervical approach in cervical spondylotic diseases. Methods A total of 75 patients were selected, who underwent microscopic subtotal anterior cervical decompres-sion and titanium mesh to fusion fixation or microscopic decompression of resection of the cervical intervertebral disc and cage implant fixation in Tianjin People's Hospital since October 2011 to October 2012. The Cervical vertebra gap involved 1 level in 41 cases, 2 levels in 30 cases and 3 levels in 4 cases. Pateints followed up regularly after operation. The nerve func-tion was assessed using Japanese orthopaedic association scoring system (JOA) function was calculated to examine the im-provement rate and to assess the curative effect of microscopic anterior cervical approach. Results All patients were fol-lowed up. The nerve function recovery was excellent in 44 cases, was well in 25 cases, was general in 6 cases, and the ex-cellent and well recovery rate was 92%. There was no significant difference between excellent recovery rate and well recov-ery rate. There were significant differences in values of each cervical levels between before and after surgery in the last fol-low-up of patients (P<0.05). Conclusion Microscopic operative treatment for cervical spondylotic is safe and effective, with less trauma and bleeding. After surgical decompression, the recovery is rapid in patients with early ambulation.

BACKGROUND:Screw loosening and shedding may occur after osteoporosis associated with lumbar degenerative disease treated with pedicle screw fixation. Application of pedicle screw enhanced with cured materials can improve the therapeutic effect.
OBJECTIVE:To compare the clinical effect of pedicle screws enhanced with poly(methyl methacrylate) and injectable calcium sulfate cement in the lumbar internal fixation of osteoporosis.
METHODS:Sixty-one patients diagnosed with osteoporosis combined with lumbar spondylolisthesis, lumbar spinal instability, and severe lumbar spinal stenosis were col ected. Al patients were divided into two groups according to the treatment method:poly(methyl methacrylate) bone cement enhanced pedicle screw group and calcium sulfate bone cement enhanced pedicle screw group.
RESULTS AND CONCLUSION:There were no significant differences in the operation time, blood loss, preoperative and postoperative visual analog scale score, Japanese Orthopedic Association scores and the Japanese Orthopedic Association score improvement rate between two groups (P>0.05). The results showed that two patients had bone cement leakage in poly(methyl methacrylate) group which had no neurological symptoms caused by new symptoms during fol ow-up period. The bone mineral density was not improved gradual y in poly(methyl methacrylate) group with fol ow-up time prolonging;however, in calcium sulfate group, the bone mineral density was increased significantly after treatment, and the change of bone mineral density was linearly related with Japanese Orthopaedic Association score improvement rate in calcium sulfate group. No screw loosening, pul ing out or neurological dysfunction occurred in both groups. The results indicate that like poly(methyl methacrylate), balcium sulfate bone cement can increase the stability of pedicle screws.

Objective To investigate the effects of traditional Chinese medicine colquhounia root tablet on the expression of tight junction protein claudin-2 and ZO-1 in bronchial epithelium tissues of rats with acute lung injury (ALI), and to study the mechanism of protective effect of colquhounia root tablet on ALI. Methods Twenty-four healthy male Sprague-Dawley (SD) rats were randomly divided into control group, ALI group and colquhounia root tablet pretreatment group, with 8 rats in each group. The model of ALI was reproduced by intravenous injection of oleic acid 0.04 mL/kg, and the rats in cont rol group were given the same amount of normal saline (NS) instead. The rats in colquhounia root tablet pretreatment group were intragastric administrated with colquhounia root tablet of 600 mg·kg-1·d-1 (2 mL) for 10 days before model reproduction, and the rats in control group and ALI group were given the same amount of NS. At 4 hours after model reproduction, the blood was drawn from abdominal aorta, and bronchoalveolar lavage fluid (BALF) was collected for determination of protein content in plasma and BALF, and the lung permeability index (LPI) was calculated. The rats were sacrificed to collect lung tissues for determination of lung wet/dry weight ratio (W/D), the changes in pathology of lung tissue were observed after hematoxylin and eosin (HE) staining with light microscope, and lung injury score (LIS) was evaluated. The immunohistochemic al staining was used to detect the expression and localization of claudin-2 and ZO-1 in bronchial epithelium tissues. The protein expressions of claudin-2 and ZO-1 in bronchial epithelium tissues were determined by Western Blot. Results Compared with control group, the lung injury in ALI group was more obvious including cellular edema and structural disorder of intercellular connection by optical microscope, and LIS, W/D ratio, and LPI were significantly increased (LIS: 3.81±0.42 vs. 0.40±0.08, W/D: 7.68±0.64 vs. 4.44±0.39, LPI: 0.89±0.15 vs. 0.38±0.05, all P < 0.01). Claudin-2 and ZO-1 were mainly expressed in the bronchial epithelium cell, and the expression degrees were significantly weakened in ALI group as compared with control group. It was shown by Western Blot results that compared with control group, the protein expressions of claudin-2 and ZO-1 were significantly down-regulated in ALI group [claudin-2 protein (gray value): 0.43±0.31 vs. 2.16±1.33, ZO-1 protein (gray value): 1.25±0.41 vs. 2.82±0.76, both P < 0.01]. Compared with ALI group, colquhounia root pretreatment could effectively diminish the degree of ALI (LIS: 1.22±0.39 vs. 3.81±0.42, W/D: 4.62±0.84 vs. 7.68±0.64, LPI: 0.46±0.07 vs. 0.89±0.15, all P < 0.01), and the protein expressions of claudin-2 and ZO-1 were significantly up-regulated [claudin-2 protein (gray value): 2.98±0.91 vs. 0.43±0.31, ZO-1 protein (gray value): 2.35±0.51 vs. 1.25±0.41, both P < 0.01]. Conclusion Administration of colquhounia root table could attenuate lung injury induced by oleic acid with improving epithelial barrier function via up-regulate the expression claudin-2 and ZO-1, which play a protective effect on the lung of rats with ALI.

Objective:To validate an enzyme linked immunosorbent assay (ELISA) method for the quantification of rhCNB in long-tailed macaque sera.Methods: The linear,sensitivity,accuracy,precision and recovery were determined using ELISA.Results:The present ELISA method had high linearity within 0.195 ng/ml-12.5 ng/ml,the working curve of rhCNB was Y=15.1X-0.26, R2=0.996 8 , the method showed good sensitivity of 0.195 ng/ml, the accuracy were in the range of 91.9%-108.8%, and the Coefficient of variation ( CV) for inter-assay were 3.55%,1.39%and 4.71%,the intra-assay were 1.59%,3.2%and 3.8%,all less than 10%, the recoveries were in the range of 88.5% -108.3%, <110% .Thus the method was coincidence with requirement.Conclusion:Double antibody sandwich ELISA assay of rhCNB in long-tailed macaque sera has good sensitivity ,accuracy, precision and recovery and it can be used to measure rhCNB concentration in biological samples .

OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

A sensitive, selective and simple liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method was developed for determining of daptomycinin human plasma and effluent. The analyte was extracted from plasma samples by SPE method, separated through a Phenomenex Kinetex C18 column(50 mm×2. 1 mm, 1. 7 μm)using isocratic mobile phase consisting of 0. 1% formic acid-acetonitile(75 ∶25), and analyzed by electro-spray ionization(ESI). The precursor to product ion transitions of m/z 810. 9→159. 1 and m/z 286. 2→217. 2 were used to measure daptomycinand the internal standard, respectively. The method was validated over a concentration range(plasma: 1-200 μg/mL, effulent: 0. 005-20 μg/mL). The intra- and inter-day precision values were less than 10% and accuracy values 90%-110%. The stability of daptomycinin human plasma and effluent under different storage conditions met the requirements of bioanalytical method. The concentration of daptomycin is significant lower in the septic shock patient, when give a dose of 6 mg/kg, the cmax and AUC0-24 h of steady state decreased by 50% and 60% respectively; the increase in capillary permeability and interstitial oedema during sepsis and septic shock may enhance drug distribution. By the way, daptomycin can be cleared via continuous veno-venous hemofiltration(CVVH)for nearly 16%. In summary, on the treatment of continuous renal replacement therapy(CRRT)in patients with septic shock with daptomycin therapy, the suggested dose should be increased, and the drug monitoring should be carried on.

As one of the first-line antipsychotic drugs, olanzapine(OLA)inducing insulin resistance and metabolic disorder has become a serious clinical concern. This study aimed at investigating the effect of OLA on white adipose tissue(WAT)beiging as a potential mechanism behind its metabolism-disturbing effects. C57BL6/J mice were orally administered with low and high-dose OLA(4, 8 mg/kg, once daily)for 28 consecutive days; body weight and food intake were recorded every other day. Mice were subjected to glucose-tolerance test(GTT)and a cohort from each group was challenged with cold stress for 36 h before sacrifice. The perirenal, epididymal and inguinal WAT were carefully dissected, weighed and processed separately for hematoxylin-eosin staining, UCP-1 immunohistochemical and Western blot analysis. The results showed that OLA induced significant impairment in basal heat generation and glucose regulation. There were fewer beige adipocytes in the inguinal WAT after OLA treatment. The beiging of inguinal WAT in response to cold stress was significantly inhibited in OLA-treated mice. Mechanistically, OLA induced the activation of mTOR-Notch pathway, as evidenced by a significant increase in phosphorylated mammalian target of rapamycin(p-mTOR)and intracellular domain of Notch1(N1ICD)expression. Together, our study reveals that the inhibitory effects on WAT beiging may explain the propensity of OLA to induce metabolic disturbance, and mTOR-Notch pathway activation could play key roles in this effect. Future validation and elucidation of these novel findings are expected to provide novel insights into the metabolic risks of OLA and related second generation antipsychotics, which may lead to innovative intervention strategies in the clinic.

Objective:To investigate the clinical value of radiomics based on computed tomography (CT) examination in preoperative differential diagnosis of pancreatic serous cystadenoma (SCA) and mucinous cystadenoma (MCA).Methods:The retrospective case-control study was conducted. The clinicopathological and imaging data of 154 patients with pancreatic cystic neoplasms who were admitted to the First Affiliated Hospital, Zhejiang University School of Medicine from January 2012 to December 2019 were collected. There were 24 males and 130 females, aged (50±13)years. Of the 154 patients, 99 cases were diagnosed as SCA and 55 cases were diagnosed as MCA. All the 154 patients underwent plain and enhanced CT scan of pancreas before operation. The clinical characteristics, radiology features and radiomics features of all patients were collected to construct the clinical characteristics model, radiology model, radiomics model and fused model. The receiver operating characteristic (ROC) curve of each model was drawn, and those constructed models were evaluated by area under the curve (AUC), accuracy, sensitivity, specificity, positive predictive value and negative predictive value. Based on the optimal model, the nomogram was constructed. Observation indicators: (1) establishment and validation of clinical characteristics model; (2) establishment and validation of radiology model; (3) establishment and validation of radiomics model; (4) establishment and validation of fused model; (5) nomogram of fused model. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Results:(1) Establishment and validation of clinical characteristics model: 3 clinical characteristics, including age, symptoms and preoperative serum CA19-9, were selected using multinomial logistic linear regression analysis to construct the clinical characteristics model. Result of the multinomial logistic linear regression analysis was expressed by formula ①: clinical characteristics model score=0.635-0.007×age+0.054×clinical symptoms+0.108×preoperative serum CA19-9. The ROC curve for the test dataset of clinical characteristics model was drawn. The AUC, accuracy, sensitivity, specificity, positive predictive value and negative predictive value of clinical characteristics model were 0.611(95% confidence interval as 0.488?0.734, P<0.05), 56.6%, 66.7%, 56.3%, 41.5%, 78.4% for the training dataset and 0.771(95% confidence interval as 0.624?0.919, P<0.05), 77.8%, 63.1%, 88.5%, 80.1%, 76.7% for the test dataset, respectively. (2) Establishment and validation of radiology model: 5 radiology characteristics, including tumor location, the number of tumors, tumor diameter of cross section, lobulated tumor and polycystic tumor (more than 6), were selected using multinomial logistic linear regression analysis to construct the radiology model. Result of the multinomial logistic linear regression analysis was expressed by formula ②: radiology model score=?0.034+0.300×tumor location+0.202×the number of tumors+0.014×tumor diameter of cross section?0.251×lobulated tumor?0.170×polycystic tumor (more than 6). The ROC curve for the test dataset of radiology model was drawn. The AUC, accuracy, sensitivity, specificity, positive predictive value and negative predictive value of radiology model were 0.862(95% confidence interval as 0.791?0.932, P<0.05), 78.8%, 81.8%, 77.5%, 62.8%, 90.2% for the training dataset and 0.853(95% confidence interval as 0.713?0.994), P<0.05), 88.9%, 89.4%, 88.5%, 85.0%, 92.0% for the test dataset, respectively. (3) Establishment and validation of radiomics model: 4 categories of a total 1 067 radiomics features were extracted from 154 patients with pancreatic cystic neoplasms, including 7 first-order histogram features, 53 texture features, 848 wavelet features and 159 local binary pattern features. A total of 896 stable radiomics features were retained to construct the model, based on the condition of intraclass correlation coefficient >0.9. After selected by variance threshold and correlation coefficient threshold, 350 radiomics features were retained. Fifty synthetic radiomics features were constructed based on the original features in order to obtain potential radiomics features, and the total number of radiomics features was 400. After analyzed by the five-fold recursive feature elimination, 22 radiomics features were screened out, including 13 wavelet features, 7 synthetic radiomics features and 2 local binary pattern features. The support vector machine algorithm was used to construct the radiomics model. The penalty coefficient 'C' and parameter 'γ' of the radiomics model were 35.938 and 0.077, respectively. The kernel function of the radiomics model was 'radial basis function kernel'. The ROC curve of radiomics model using 5-fold cross validation was drawn. The average AUC, accuracy, sensitivity, specificity, positive predictive value and negative predictive value of the radiomics model were 0.870 ( P<0.05), 83.1%, 81.8%, 83.8%, 73.8% and 89.2%, respectively. (4) Establishment and validation of fused model: the fused model was constructed after selecting the tumor location and lobulated tumor of radiology characteristics and radiomics score. Result of the multinomial logistic linear regression analysis was expressed by formula ③: fused model socre=?0.154+0.218×tumor location?0.223×lobulated tumor+0.621×radiomics score. The ROC curve for the test dataset of fused model was drawn. The AUC, accuracy, sensitivity, specificity, positive predictive value and negative predictive value of fused model were 0.893(95% confidence interval as 0.828?0.958, P<0.05), 83.7%, 81.8%, 84.5%, 71.1%, 90.9% for the training dataset and 0.966(95% confidence interval as 0.921?0.999, P<0.05), 91.1%, 84.2%, 96.2%, 94.1%, 89.3% for the test dataset, respectively. (5) Nomogram of fused model: the nomogram of fused model was illustrated with the Youden index of 0.416. Conclusion:The prediction model based on the radiomics signature and radiological features extracted from preoperative CT examination can make the differential diagnosis of pancreatic SCA from MCA.

Objective:To discuss the stability of mixed standard application solution by volatilization organic matter (benzene, toluene, xylene, ethylbenzene, acetone, butanone, Ethyl acetate, butyl ester, trichloroethylene) in workplace air.Methods:A total of 11 kinds volatilization organic compounds were separated by capillary chromatographic column, and detected with flame ionization detector. The stability of mixed standard application solution was judged by studying three paratemers during the placement period, such as the linear correlation coefficient of the standard curve, the relative response values of the highest and lowest concentration levels and the accuracy of the measured values of the quality control samples.Results:Within 187 days, the linear correlation coefficient of each compound was ≥0.999. The changes of relative response values at the highest and lowest concentration levels were both <10%; The measured values of toluene were all in the reference range.Conclusion:The mixed standard application solution is stable and reliable within 187 days of storage.

OBJECTIVE: To analyze the clinical phenotype and MYH7 gene variant in a Chinese pedigree affected with hypertrophic cardiomyopathy (HCM). METHODS: The proband was screened for variant of 96 cardiomyopathy-associated genes by exonic amplification and high-throughput sequencing. Candidate variant was verified by Sanger sequencing among 300 healthy controls as well as family members of the proband. Co-segregation analysis of genotypes and clinical phenotypes was carried out for the pedigree. Clustal X software was used to analyze the sequence conservation of the variant among various species, and its pathogenicity was predicted by using bioinformatics software. RESULTS: 6 out of 12 members from this pedigree were found to harbor heterozygous c.4124A>G (p.Tyr1375Cys) variant of the MYH7 gene, among whom five were diagnosed with HCM. The remaining one had failed to meet the diagnostic criteria for HCM, but had abnormal ECG. The same variant was not found in the 300 healthy controls. Amino acid sequence analysis showed that the variant is located in a highly conserved region, and bioinformatics analysis predicted that this variant may affect protein function and has a deleterious effect. Based on the American College of Medical Genetics and Genomics (ACMG) guidelines, the variant was predicted to be likely pathogenic (PM2+ PP1_Moderate+PP3+PP5). CONCLUSION The c.4124A>G (p.Tyr1375Cys) variant of the MYH7 gene probably underlay the pathogenesis in this pedigree. Above finding has important value for the early diagnosis of patients with HCM.
Cardiac Myosins/genetics* ; Cardiomyopathy, Hypertrophic/genetics* ; Genotype ; Humans ; Mutation ; Myosin Heavy Chains/genetics* ; Pedigree ; Phenotype