Naringenin attenuates cerebral ischemia/reperfusion injury by inhibiting oxidative stress and inflammatory response via the activation of SIRT1/FOXO1 signaling pathway in vitro

Purpose: To explore the protection of naringenin against oxygen-glucose deprivation/reperfusion (OGD/R)-induced HT22 cell injury, a cell model of cerebral ischemia/reperfusion (I/R) injury in vitro, focusing on SIRT1/FOXO1 signaling pathway. Methods: Cytotoxicity, apoptosis, reactive oxygen species (ROS) generation, malondialdehyde (MDA) content, 4-hydroxynonenoic acid (4-HNE) level, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities were measured by commercial kits. Inflammatory cytokines levels were determined by enzyme-linked immunosorbent assay (ELISA). The protein expressions were monitored by Western blot analysis. Results: Naringenin significantly ameliorated OGD/Rinduced cytotoxicity and apoptosis in HT22 cells. Meanwhile, naringenin promoted SIRT1 and FOXO1 protein expressions in OGD/R-subjected HT22 cells. In addition, naringenin attenuated OGD/R-induced cytotoxicity, apoptosis, oxidative stress (the increased ROS, MDA and 4-HNE levels, and the decreased SOD, GSH-Px and CAT activities) and inflammatory response (the increased tumor necrosis factor-α, interleukin [IL]-1ß, and IL-6 levels and the decreased IL-10 level), which were blocked by the inhibition of the SIRT1/FOXO1 signaling pathway induced by SIRT1-siRNA transfection. Conclusion: Naringenin protected HT22 cells against OGD/R injury depending on its antioxidant and anti-inflammatory activities via promoting the SIRT1/FOXO1 signaling pathway.

Association Between Antibiotic Overexposure and Adverse Outcomes in Very-Low-Birth-Weight Infants Without Culture-Proven Sepsis or Necrotizing Enterocolitis: A Multicenter Prospective Study

Objectives To explore the associations between higher antibiotic use rates (AURs) and adverse outcomes in very-low-birthweight (VLBW) infants without culture-proven sepsis or necrotizing enterocolitis (NEC) in a multicenter of China. Methods A prospective cohort study was performed on VLBW infants admitted to 24 neonatal intensive care units from January 1, 2018, to December 31, 2018. AUR was calculated as calendar days of antibiotic therapy divided by total hospital days. The composite primary outcome was defned as mortality or severe morbidity, including any of the following: severe neurologic injury, bronchopulmonary dysplasia (BPD), and stage 3 or higher retinopathy of prematurity. Results A total of 1,034 VLBW infants who received antibiotics without culture-proven sepsis or NEC were included in this study. The overall AUR of eligible VLBW infants was 55%, and the AUR of each eligible VLBW infant ranged from 3 to 100%, with a median of 56% (IQR 33%, 86%). After generalized propensity score and logistic regression analysis of 4 groups of VLBW infants with diferent AUR range, infants in the higher quartile AUR, (Q3, 0.57~0.86) and (Q4, 0.87~1.00), had higher odds of composite primary outcome (adjusted OR: 1.81; 95% CI: 1.23–2.67; adjusted OR 2.37; 95% CI: 1.59–3.54, respectively) and BPD (adjusted OR: 3.09; 95% CI: 1.52–6.57; adjusted OR 3.17; 95% CI: 1.56–6.57, respectively) than those in the lowest AUR (Q1). Conclusions Antibiotic overexposure in VLBW infants without culture-proven sepsis or NEC was associated with increased risk of composite primary outcome and BPD. Rational empirical antibiotic use in VLBW infants is urgently needed in China.

Personalized quantitative evaluation of the quality of radiotherapy plans based on dose prediction / 中华放射医学与防护杂志

Objective:To develop a dose prediction-based quantitative evaluation method of the quality of radiotherapy plans, and to verify the clinical feasibility and clinical value of the method .Methods:The 3D U-Netwas trained using the radiotherapy plans of 45 rectal cancer cases that were formulated by physicists with more than five years of radiotherapy experience. After obtaining 3D dose distribution using 3D U-Net prediction, this study established the plan quality metrics of intensity modulated radiotherapy(IMRT) rectal cancer radiotherapy plans using dose-volume histogram(DVH) indexes of dose prediction. Then, the initial scores of rectal cancer radiotherapy plans were determined.Taking the predicted dose as the optimization goal, the radiotherapy plans were optimized and scored again. The clinical significance of this scoring method was verified by comparing the scores and dosimetric parameters of the 15 rectal cancer cases before and after optimization.Results:The radiotherapy plans before and after optimization all met the clinical dose requirements. The total scores were(77.21±9.74) before optimization, and (88.78±4.92) after optimization. Therefore, the optimized radiotherapy planswon increased scores with a statistically significant difference( t=-4.105, P<0.05). Compared to the plans before optimization, the optimized plans show decreased Dmax of all organs at risk to different extents. Moreover, the Dmax, V107%, and HI of PTV and the Dmax of the bladder decreased in the optimized plans, with statistically significant differences ( t=2.346-5.771, P<0.05). There was no statistically significant difference in other indexes before and after optimization ( P>0.05).The quality of the optimized plans were improved to a certain extent. Conclusions:This study proposed a dose prediction-based quantitative evaluation method of the quality of radiotherapy plans. It can be used for the effective personalized elevation of the quality of radiotherapy plans, which is beneficial to effectively compare and review the quality of clinical plans determined by different physicists and provide personalized dose indicators. Moreover, it can provide great guidance for the formulation of clinical therapy plans.

Wutou Chishizhi Wan Regulates Autophagy and PI3K/Akt/GSK-3β Signaling Pathway in Rats with Myocardial Ischemia-reperfusion Injury / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Wutou Chishizhi Wan in regulating autophagy and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/glycogen synthase kinase-3β （GSK-3β） signaling pathway in rats with myocardial ischemia-reperfusion injury （MIRI）. MethodSixty male SD rats were randomly assigned into the normal group （normal saline）， model group （normal saline）， positive control （trimetazidine， 5.4 mg·kg-1） group， and low-， medium-， and high-dose （1.63， 4.9， 14.7 g·kg-1， respectively） Wutou Chishizhi Wan groups， with 10 rats in each group. The rats in other groups except the normal group underwent left anterior descending coronary artery ligation for modeling. Electrocardiogram was employed to detect the ST-segment elevation to evaluate the modeling. Hematoxylin-eosin （HE） staining was performed to reveal the damage of myocardial tissue. The levels of aspartate aminotransferase （AST） and creatine kinase （CK） were determined by colorimetry， and those of cardiac troponin T （cTnT） and myoglobin （MYO） by enzyme-linked immunosorbent assay （ELISA）. Western blot was carried out to determine the protein levels of microtubule-associated proteins 1 light chain 3 （LC3）， autophagy-related gene Beclin-1， PI3K， Akt， GSK-3β， p-GSK-3β， and p-Akt. ResultCompared with the normal group， the modeling elevated the serum levels of AST， CK， cTnT， and MYO （P<0.01）， destroyed the arrangement of myocardial cells abd nucle， twisted and broken myocardial fibers， up-regulated the protein levels of LC3Ⅱ/Ⅰ and Beclin-1 （P<0.01）， and down-regulated the protein levels of PI3K， p-Akt， and p-GSK-3β （P<0.01）. Compared with the model group， trimetazidine and Wutou Chishizhi Wan （all the doses） lowered the levels of AST， CK， cTnT， and MYO in serum （P<0.01）， restored the arrangement of myocardial cells and muscle fibers， reduced necrosis， down-regulated the protein level of Beclin-1 （P<0.01）， and up-regulated the protein levels of PI3K， p-Akt， and p-GSK-3β （P<0.01）. Additionally， Wutou Chishizhi Wan （all the doses） down-regulated the protein level of LC3Ⅱ/Ⅰ （P<0.05， P<0.01）. Compared with those in the trimetazidine group， the serum AST level rose in the low-dose Wutou Chishizhi Wan group （P<0.05） and declined in the high-dose group （P<0.01）， and the protein level of Beclin-1 was down-regulated in the medium-dose group （P<0.01）. Additionally， the trimetazidine group had higher protein level of LC3Ⅱ/Ⅰ than medium- and high-dose Wutou Chishizhi Wan groups （P<0.05， P<0.01）， higher protein level of PI3K than low-， medium-， and high-dose groups （P<0.01）， lower protein level of p-Akt than low- and medium-dose groups （P<0.01）， and higher p-GSK-3β protein level than the medium-dose group （P<0.01）. ConclusionDifferent doses of Wutou Chishizhi Wan can ameliorate MIRI， and the high dose has the best effect. Wutou Chishizhi Wan can reduce the activity of myocardial injury markers AST， CK， cTnT， and MYO， and alleviate the pathological damage of myocardial tissue. It can down-regulate the protein levels of beclin-1， LC3Ⅱ/Ⅰ， and up-regulate those of PI3K， p-Akt， and p-GSK-3β. In summary， Wutou Chishizhi Wan may inhibit excessive autophagy and regulate the PI3K/Akt/GSK-3β signaling pathway to exert protective effect on MIRI rats.

Comparison of two prediction models for mortality ofacute paraquat poisoning / 预防医学

Objective@#To compare the effects of Cox proportional hazard regression model (Cox model) and extreme gradient boosting model ( XGBoost model ) on the prediction of the mortality of acute paraquat poisoning (APP).@*Methods@#The APP cases admitted to Qingdao Eighth People's Hospital and Shandong Provincial Hospital from January 1st of 2018 to December 1st of 2020 was recruited and divided into a training group and a verification group by a random number table. The Cox model and XGBoost model were established to select the predictors for APP mortality. Receiver operating characteristic ( ROC ) curve was drawn to analyze the predictive power of the two models, and the calibration was evaluated using Hosmer-Lemeshow test.@*Results@#Totally 150 APP cases were recruited. There were 75 cases each in the training group and in the verification group, with 52 and 55 cases died respectively, accounting for 69.33% and 73.33%. The Cox model showed that paraquat intake, the time from taking poison to seeing a doctor, the time for the first perfusion, the time for the first vomiting, aspartate aminotransferase, alanine aminotransferase, serum creatinine, blood urea nitrogen, white blood cell, lactic acid, creatine kinase isoenzymes, glucose, serum calcium and serum potassium were the predictors of APP mortality ( all P<0.05 ). The XGboost model showed that the predictive power of the factors in a descending order were the time from taking poison to seeing a doctor, the time for the first vomiting, the time for the first perfusion, lactic acid, white blood cell, paraquat intake, serum creatinine, serum potassium, serum calcium, creatine kinase isoenzymes, glucose, aspartate aminotransferase, blood urea nitrogen and alanine aminotransferase. The area under curve ( AUC ) of the XGBoost model for predicting was 0.972, which was greater than 0.921 of the Cox model ( P<0.05 ). The predicted results of the Cox model and XGBoost model were consistent with the actual situation ( P>0.05 ). @*Conclusion@#The Cox model and XGBoost model are consistent in predicting the mortality of APP, but the latter is better.

Epidemiological characterization of sapovirus gastroenteritis aggregation in a kindergarten in Jinshan District， Shanghai / 上海预防医学

Objective:To analyze the epidemiological characteristics of an aggregational gastroenteritis and determine the genotypes of sapovirus， and to provide scientific basis for formulating effective control strategies. Methods:Unified case definition， active case search and descriptive epidemiology were used to analyze the epidemic. Feces or anal swabs of untreated students， teachers， canteen staff as well as canteen environment samples were collected. Norovirus and sapovirus nucleic acid tests were conducted by real-time fluorescent RT-PCR， and sapovirus nucleic acid was amplified by conventional RT-PCR. The gene region of capsid protein was analyzed by MEGA7.0 software and phylogenetic tree was constructed. Results:A total of 12 cases were reported in the epidemic， and the incidence rate was 44.44%. All reported cases， with vomiting symptoms， were found in the same class. The epidemic showed a point-based outbreak. The first case became the source of infection in class， and the epidemic lasted for 8 days. Real-time fluorescent RT-PCR assay confirmed that five children's feces were positive for sapovirus nucleic acid， and the first-episode children's feces were positive for sapovirus and GII norovirus nucleic acid. Sequence alignment result showed that the sapovirus strains belonged to GI.1 type with homologous genes. Conclusion:Based on the clinical manifestations， field epidemiological investigation and laboratory test results， we confirm that the first case of the epidemic in class is caused by GI.1 sapovirus infection. The epidemic is effectively controlled by comprehensive measures such as case isolation and disinfection.

Dose distributions prediction for intensity-modulated radiotherapy of postoperative rectal cancer based on deep learning / 中华放射医学与防护杂志

Objective:To develop a deep learning model for predicting three-dimensional (3D) voxel-wise dose distributions for intensity-modulated radiotherapy (IMRT).Methods:A total of 110 postoperative rectal cancer cases treated by IMRT were considered in the study, of which 90 cases were randomly selected as the training-validating set and the remaining as the testing set. A 3D deep learning model named 3D U-Res-Net was constructed to predict 3D dose distributions. Three types of 3D matrices from CT images, structure sets and beam configurations were fed into the independent input channel, respectively, and the 3D matrix of IMRT dose distributions was taken as the output to train the 3D model. The obtained 3D model was used to predict new 3D dose distributions. The predicted accuracy was evaluated in two aspects: the average dose prediction bias and mean absolute errors (MAEs)of all voxels within the body, the dice similarity coefficients (DSCs), Hausdorff distance(HD 95) and mean surface distance (MSD) of different isodose surfaces were used to address the spatial correspondence between predicted and clinical delivered 3D dose distributions; the dosimetric index (DI) including homogeneity index, conformity index, V50, V45 for PTV and OARs between predicted and clinical truth were statistically analyzed with the paired-samples t test. Results:For the 20 testing cases, the average prediction bias ranged from -2.12% to 2.88%, and the MAEs varied from 2.55% to 5.75%. The DSCs value was above 0.9 for all isodose surfaces, the average MSD ranged from 0.21 cm to 0.45 cm, and the average HD 95 varied from 0.61 cm to 1.54 cm. There was no statistically significant difference for all DIs, except for bladder Dmean. Conclusions:This study developed a deep learning model based on 3D U-Res-Net by considering beam configurations input and achieved an accurate 3D voxel-wise dose prediction for rectal cancer treated by IMRT.

Protective Effect of Wutou Chishizhi Wan on Vascular Endothelial Cells and Oxidative Stress in Myocardial Ischemia-reperfusion Rats / 中国实验方剂学杂志

Objective:To investigate the protective effect of Wutou Chishizhi Wan on myocardial ischemia reperfusion injury （MIRI） in rats， and observe its effect on such mechanisms as coagulation function， vascular endothelial cells and oxidative stress in rats. Method:A total of 40 SD rats were randomly divided into normal group， model group， positive drug group （Urokinase group） and Wutou Chishizhi Wan group， with 10 rats in each group. Except for the normal group， rat myocardial ischemia-reperfusion injury models were established. The changes of heart rate （HR） at 10 min before ischemia， 30 min after ischemia and 30， 60， 120 min （T0，T1，T2，T3，T4）， and the change of electrocardiogram （ECG） J point after modeling in rats were observed. The pathological changes of rat myocardial tissue were observed by hematoxylin-eosin （HE） staining. The changes of four indexes of coagulation ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， thrombin time （TT）， fibrinogen content decreased significantly （FIB）］ in rats were observed. The contents of endothelin-1 （ET-1）， thromboxane A2 （TXA2） and prostacyclin （PGI2） in serum and myocardium levels of superoxide dismutase （SOD）， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） of MIRI rats were observed. Western blot assay was used for the detection of oxidative stress protein Keap1 and transcription factor-E2-related factor （Nrf2） expression levels in rat myocardial tissue. Result:Compared with the normal group， the ECG of MIRI rats showed significant myocardial ischemic injury-like changes， ST segment was significantly elevated， J point was significantly increased， and the incidences of HR in T1， T2， T3 and T4 were significantly reduced （P<0.05， P<0.01）. Compared with the model group， Wutou Chishizhi Wan significantly reduced ECG J-point changes in MIRI rats， while increased the incidence of HR in T1， T2， T3 and T4 （P<0.05， P<0.01）. Compared with the normal group， PT， APTT and TT in the model group were significantly shortened （P<0.01）， FIB content was significantly increased （P<0.01）， and the serum PGI2 level decreased and TXA2 and ET-1 levels increased significantly in the model group （P<0.01）. SOD content and GSH-Px activities of myocardial tissue in the model group were significantly reduced （P<0.01）， whereas the MDA content was increased （P<0.01）. Compared with the model group， PT of the Wutou Chishizhi Wan group was prolonged （P<0.05） and APTT slightly prolonged， TT significantly prolonged （P<0.01）， FIB content decreased （P<0.05）， serum PGI2 increased （P<0.05）， TXA2 and ET-1 decreased significantly in the Wutou Chishizhi Wan group （P<0.01）， myocardial MDA content decreased， and SOD content and GSP-Px activity increased significantly （P<0.01）. Meanwhile， the Wutou Chishizhi Wan group was able to activate the Keap1/Nrf2 signaling pathway， which significantly increased Nrf2 expression and significantly decreased Keap1 expression （P<0.01）. Conclusion:Wutou Chishizhi Wan group can protect myocardial injury in MIRI rats. The specific mechanism is to protect MIRI by regulating vascular endothelial cell homeostasis and oxidative stress levels and activating Keap1/Nrf2 signaling pathway.

A survey on therapy strategies for rheumatoid arthritis in Chinese rheumatologists / 中华内科杂志

To investigate how Chinese rheumatologists treated patients with rheumatoid arthritis (RA). We performed a survey on the choices of first-line and second-line anti-RA therapies, prescription of methotrexate and glucocorticoids, assessment of disease activity and frequencies of follow-up at the Asia Pacific League of Associations for Rheumatology meeting 2016 in Shanghai. The majority (85.1%) of rheumatologists preferred methotrexate as first-line treatment. As alternative agents, 71.0% rheumatologists chose leflunomide or sulfasalazine. If methotrexate was not tolerable, only 8.6% rheumatologists would switch to parenteral administration. After failure of responding to methotrexate, 62.0% rheumatologists recommended to change or combine other conventional synthetic disease modifying anti-rheumatic drugs (DMARDs). Etanercept was the most popular biological option in 65.2% rheumatologists. Almost all (97.3%) rheumatologists prescribed methotrexate at an initial dose of 7.5 to 15 mg/week and 73.8% rheumatologists at a maximum of 10 to 15 mg/week. There were 49.3% rheumatologists prescribing oral glucocorticoids at first-line therapy. Surprisingly, 42.6% rheumatologists never or rarely assessed disease activity in daily work. For patients having achieved remission, 74.2% rheumatologists would follow up them every 1 to 3 months. This study suggests that most Chinese rheumatologists treat RA patients consistent with international guidelines, while the maximum dose of methotrexate, glucocorticoid as first-line treatment, assessment of disease activity and follow-up frequency are locally modified.

Stature Estimation of Sichuan Han Females Based on X-ray Photography of Total Lower Limbs / 法医学杂志

OBJECTIVES@#To establish a mathematical model of stature estimation for Sichuan Han females on the basis of the relationship between lower limbs and individual height, thus to provide evidence for forensic identification.@*METHODS@#Samples were collected from 171 Sichuan Han females. Large flat panel multi-function digital photography system was used to take the full-body X-ray films of the lower limbs. Indexes of long bones and stature of the subjects were measured, respectively. A linear regression analysis was carried out on the correlation between them, and a mathematical model of the stature calculation was established. Then the mathematical model was used to calculate the stature of another 29 Sichuan Han females to test its accuracy.@*RESULTS@#The maximum length of femur （x1） had the highest correlation with stature. A total of 13 linear regression equations were established （P<0.05）, with the correlation coefficient （R） 0.821-0.897 and the standard error of the estimation （SEE） 2.994-3.812 cm. The backtesting showed that the equation y=41.604+1.205 x1+1.318 x6+2.444 x12+1.852 x13-2.388 x14 had the smallest mean absolute deviation （2.485 years old） and the highest accuracy of ±2SEE （92.9%）, and that the equation y=48.783+2.568 x1 had the highest accuracy of ±1SEE （60.7%）.@*CONCLUSIONS@#The stature estimation is high by using the long bones of the lower limbs has high accuracy.

Serum regucalcin is a useful indicator of liver injury severity in patients with hepatitis B virus-related liver diseases

Regucalcin is a soluble protein that is principally expressed in hepatocytes. Studies of regucalcin have mainly been conducted in animals due to a lack of commercially available kits. We aimed to develop an enzyme-linked immunosorbent assay (ELISA) to quantify serum regucalcin in patients with hepatitis B virus (HBV)-related disease. High-titer monoclonal antibodies and a polyclonal antibody to regucalcin were produced, a double-antibody sandwich ELISA method was established, and serum regucalcin was determined in 47 chronic hepatitis B (CHB) patients, 91 HBV-related acute-on-chronic liver failure (HBV-ACLF) patients, and 33 healthy controls. The ELISA demonstrated an appropriate linear range, and high levels of reproducibility, sensitivity, specificity, accuracy, and stability. The median serum regucalcin concentrations in HBV-ACLF and CHB patients were 5.46 and 3.76 ng/mL, respectively (P<0.01), which were much higher than in healthy controls (1.72 ng/mL, both P<0.01). For the differentiation of CHB patients and healthy controls, the area under curve (AUC) was 0.86 with a cut-off of 2.42 ng/mL, 85.7% sensitivity, and 78.8% specificity. In contrast, the AUC of alanine aminotransferase (ALT) was lower (AUC=0.80, P=0.01). To differentiate ACLF from CHB, the AUC was 0.72 with a cut-off of 4.26 ng/mL, 77.0% sensitivity, and 61.2% specificity while the AUC of ALT was 0.41 (P=0.07). Thus, we have developed an ELISA that is suitable for measuring serum regucalcin and have shown that serum regucalcin increased with the severity of liver injury due to HBV-related diseases, such that it appears to be more useful than ALT as a marker of liver injury.

The study of automatic treatment planning of prostate cancer based on DVH prediction models of organs at risk / 中华放射肿瘤学杂志

Objective To evaluate the feasibility of utilizing dose-volume histogram (DVH) prediction models of organs at risk (OARs) to deliver automatic treatment planning of prostate cancer.Methods The training set included 30 cases randomly selected from a database of 42 cases of prostate cancer receiving treatment planning.The bladder and rectum were divided into sub-volumes (Ai) of 3 mm in layer thickness according to the spatial distance from the boundary of planning target volume (PTV).A skewed normal Gaussian function was adopted to fit the differential DVH of Ai,and a precise mathematical model was built after optimization.Using the embedded C++ subroutine of Pinnacle scripa,ahe volume of each Ai of the remaining validation set for 12 patients was obtained to predict the DVH parameters of these OARa,ahich were used as the objective functions to create personalized Pinnacle script.Finalla,automatic plans were generated using the script.The dosimetric differences among the original clinical plannina,aredicted value and the automatic treatment planning were statistically compared with paired t-test.Results DVH residual analysis demonstrated that predictive volume fraction of the bladder and rectum above 6 000 cGy were lower than those of the original clinical planning.The automatic treatment planning significantly reduced the V70,V60,V50 of the bladder and the V70 and V60 of the rectum than the original clinical planning (all P＜0.05),the coverage and conformal index (CI) of PTV remained unchangea,and the homogeneity index (HI) was slightly decreased with no statistical significance (P＞ 0.05).Conclusion The automatic treatment planning of the prostate cancer based on the DVH prediction models can reduce the irradiation dose of OARs and improve the treatment planning efficiency.

Application progress of new detection techniques for peripheral blood circulating tumor cells in prostate cancer / 肿瘤研究与临床

Circulating tumor cells (CTC) refer to tumor cells that survive from the primary or metastatic tumors through active or passive blood entry and escape immune killing. With the continuous development of modern detection technology, the studies on some new CTC detection and separation technology including Cell Search system, Adna Test system, high-throughput imaging platform and microfluidic chip technology in prostate cancer have gradually made in-depth progresses. This article reviews the latest application progress of these new detection techniques in prostate cancer.

Stature Estimation for Sichuan Han Female Based on Whole-spine X-ray Photo-graphy / 法医学杂志

Objective To establish regression models of stature estimation for Sichuan Han female by the measurement of total vertebral column length in the frontal and lateral X-ray films of whole-spine. Methods The frontal and lateral X-ray films of whole-spine were collected from 200 Sichuan Han fe-males by large flat-panel multi-functional universal radiography and fluoroscopy system. The data and mean values of frontal and lateral total vertebral column length were measured and calculated in all the samples, respectively. The relationship of combined multi-markers and stature were analysed by linear regression analysis, and the mathematical models of stature estimation were established. The data of 30 new samples were selected and inputted for verifying the accuracy of the mathematical models. Results The total vertebral column length showed a good correlation with stature, and the mean values of the frontal and lateral X-ray films of total vertebral column length had the highest correlation coefficients. Three established linear regression equation models were statistically significant(P＜0.05), and the equa-tion established with lateral total vertebral column length showed the highest accuracy. Conclusion The stature estimation by the measurement of total vertebral column length has high accuracy.

Age estimation in the Living from the knee by using MRI T2 weighting images with fat suppression / 中国法医学杂志

Objective The purpose of this study is to estimate the living age by MRI T2-FS images of the knee and to establish a new age estimation method without radiation in Sichuan Hans population. Methods We retrospectively evaluated sagittal T2-weighted, fat-suppression and turbo spin-echo sequence taken upon MRI of 324patients (170 males, 154 females; age 10~30) using a six-stage method. The gender difference was tested by Mann-Whitney U and the correlation between the knee and age height was tested by Spearman correlation coefficient. Regression models were built for age estimation in both genders. Results The correlation between the distal femur and age was 0.687 in males and 0.661in females and was 0.684 in males and 0.488 in females between the proximal tibia and age. Comparison of male and female revealed nonsignificant differences in the ages at the stages 1~3, 5, 6 of the distal femoral epiphysis and stage 1~3, 5 of the proximal tibial epiphysis. The fusion of distal femur commenced at 18.42 years in males and 19.36 years in females. The fusion of proximal tibia commenced at 16.93 years in males and14.68 years in females. The test of accuracy showed MAD=2.90 years in the males and MAD=3.30 years in the females in the compound regression model. Conclusion MRI T2-FS images of the knee can be an indicator for age estimation in the living and stage 6 of the distal femur can be used to determine 18-year limit.

Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities

Abstract Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high purity DNA from soil microbiota, five prewashing agents were compared in terms of their efficiency and effectiveness in removing soil contaminants. Residual contaminants were precipitated by adding 0.6 mL of 0.5 M CaCl2. Four cell lysis methods were applied to test their compatibility with the pretreatment (prewashing + Ca2+ flocculation) and to ultimately identify the optimal cell lysis method for analyzing fungal communities in forest soils. The results showed that pretreatment with TNP + Triton X-100 + skim milk (100 mM Tris, 100 mM Na4P2O7, 1% polyvinylpyrrolidone, 100 mM NaCl, 0.05% Triton X-100, 4% skim milk, pH 10.0) removed most soil humic contaminants. When the pretreatment was combined with Ca2+ flocculation, the purity of all soil DNA samples was further improved. DNA samples obtained by the fast glass bead-beating method (MethodFGB) had the highest purity. The resulting DNA was successfully used, without further purification steps, as a template for polymerase chain reaction targeting fungal internal transcribed spacer regions. The results obtained by terminal restriction fragment length polymorphism analysis indicated that the MethodFGB revealed greater fungal diversity and more distinctive community structure compared with the other methods tested. Our study provides a protocol for fungal cell lysis in soil, which is fast, convenient, and effective for analyzing fungal communities in forest soils.

Clinical significance of expression of proliferating cell nuclear antigen and 8-hydroxydeoxyguanosine in gastric carcinoma / 实用医学杂志

Objective To investigate the expression of proliferating cell nuclear antigen ( PCNA) and the level of 8-hydroxydeoxyguanosine (8-OhdG) in gastric carcinoma and their clinical significance. Methods PCNA protein expression of 63 tumor tissues , 36 paracancerous tissues and 9 chronic gastritis mucosa were ana-lyzed by immunohistochemistry and 8-O HdG of hydrolysates was determined by HPLC. Results The expression of PCNA and the level of 8-OHdG were upregulated in chronic gastrits mucosa adjacent tissues and gastric carci-noma raised gradually and these two metrics were associated with tumor differentiation and TNM stage. Conclu-sions Up-regulation of PCNA protein expression and 8-O HdG levels are observed in gastric carcinoma. Detec-tion of PCNA protein expression and 8-O HdG levels can be used to evaluate malignant biological behavior and prognosis of gastric cancer.

Mechanism of Polypeptide Extract from Scorpion Venom Combined Rapamycin in Enhancing Autophagy of H22 Hepatoma Cells: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe enhanced effects of polypeptide extract from scorpion venom (PESV) combined Rapamycin on autophagy of H22 hepatoma cells in mice and to explore its possible mechanism.</p><p><b>METHODS</b>The H22 hepatocarcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., the control group,the high dose PESV group, the low dose PESV group, and the combination group (high dose PESV + Rapamycin), 10 in each group. Mice in high and dose PESV groups were administered with 20 mg/kg and 10 mg/kg PESV respectively by gastrogavage. Mice in the combination group were administered with 2 mg/kg rapamycin and 20 mg/kg PESV by gastrogavage. The intervention lasted for 14 successive days. The tumor volume was measured once every other day, the tumor growth curve was drawn, and then the tumor inhibitory rate calculated. Pathological changes of the tumor tissue were observed by HE staining. Protein expression levels of mammal target of rapamycin (mTOR), UNC-51-like kinase-1 (ULK1), microtubule-associated protein1 light chain3 (MAPILC3A), and Beclin1 were detected by immunohistochemical assay.</p><p><b>RESULTS</b>The growth of H22 hepatoma transplantation tumor was inhibited in high and low dose PESV groups and the combination group (P < 0.05). And there was statistical difference in tumor weight and tumor volume between the combination group and high and low dose PESV groups (P < 0.05). There was no statistical difference in tumor weight or tumor volume between the high dose PESV group and the low dose PESV group (P > 0.05). lmmunohistochemical assay showed that the protein expression of mTOR was higher, but protein expressions of ULK1, MAP1LC3A, Beclin1 were lower in the control group than in the rest 3 groups (P < 0.05, P < 0.01). Compared with the high dose PESV group, protein expressions of ULK1, MAP1LC3A, and Beclin1 were obviously lower (P < 0.05).</p><p><b>CONCLUSION</b>PESV combined Rapamycin might inhibit the development of H22 hepatoma transplantation tumor in mice possibly through inhibiting the activity of mTOR, enhancing expressions of ULK1, MAP1LC3A, and Beclin1.</p>

Clinical efficacy of Paroxetine combined with mid-frequency electrical pulse acupoint stimulation for premature ejaculation / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the clinical value of Paroxetine combined with mid-frequency electrical pulse acupoint stimulation (EPAS) in the treatment of premature ejaculation (PE).</p><p><b>METHODS</b>Totally 69 PE patients were equally assigned to receive oral Paroxetine 20 mg/d, mid-frequency EPAS, or oral Paroxetine 10 mg/d combined with mid-frequency EPAS (P + EPAS) , all for 8 weeks. We obtained the intravaginal ejaculation latency time (IELT) and Chinese Index of Premature Ejaculation (CIPE-5) scores of the patients before and after treatment, and compared adverse reactions among the three groups of patients.</p><p><b>RESULTS</b>One patient of the Paroxetine group gave up treatment because of abdominal pain and nausea. Compared with the baseline, the patients in the Paroxetine, EPAS, and P + EPAS groups all showed markedly increased IELT ([0.92 ± 0.11] vs [4.07 ± 0.11] min, P < 0.01; [0.92 ± 0.12] VS [2.78 ± 0.17] min P < 0.05; [0.91 ± 0.09] vs [5.31 ± 0.13], P < 0.01) and decreased CIPE-5 scores (12.5 ± 3.0 vs 22.0 ± 2.1, P < 0.01; 12.8 ± 2.9 vs 19.5 ± 1.9, P > 0.05; 13.1 ± 2.8 vs 25.2 ± 2.1, P 0.01), with statistically significant differences between the P + EPAS group and the other two (P < 0.05). The total effectiveness rate was 95.7% in the P + EPAS group, remarkably higher than in the Paroxetine (72.7%, P < 0.05) and the EPAS group (47.8, P < 0.01).</p><p><b>CONCLUSION</b>Oral Paroxetine combined with mid-frequency EPAS has a higher safety and efficacy than either Paroxetine or EPAS alone in the treatment of PE.</p>

Study on the mechanism of polypeptide extract from scorpion venom on inhibition of angiogenesis of H 22 hepatoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the mechanism of polypeptide extract from scorpion venom (PESV) on inhibiting angiogenesis.</p><p><b>METHODS</b>The H22 hepatoma tumor model was established by subcutaneously implanting H22 hepatoma cells into mice. The tumor-bearing mice were randomly divided into 4 groups, i.e., the control group, the high dose PESV group, the low dose PESV group, and the 5-fluorouracil (5-Fu) group, 10 mice in each group. The intervention was lasted for 14 days. The growth curve of the tumor volume was drawn and the inhibition rate calculated. Pathological changes of the tumors were observed by HE staining. The microvessel density (MVD) was detected using SP method. The protein expression levels of phosphatidylinositol 3-kinase (P13K), phosphoprotein kinase B (P-Akt), hypoxia-inducible factor-1 alpha (HIF-1 )alpha, and vascular endothelial growth factor-A (VEGF-A) were detected by immunohistochemical assay and Western blot.</p><p><b>RESULTS</b>The tumor inhibitory rate was 64.8%, 43.7%, and 32.4% in the 5-Fu group, the high dose PESV group, and the low dose PESV group. Compared with the control group, the protein expression of PI3K, P-Akt, HIF-1alpha, and VEGF-A were obviously inhibited by PESV and 5-Fu (P <0. 05,P <0. 01). The MVD also decreased in the high and low dose PESV groups (P < 0.05).</p><p><b>CONCLUSIONS</b>PESV could inhibit the angiogenesis of H22 hepatoma. The mechanisms might be associated with suppressing the expression of PI3K, P-Akt, HIF-1 alpha, and VEGF-A.</p>