Objective To evaluate a new method for the isolation of rat pancreatic stellate cells (PSCs) and to investigate the influence of adipose derived stem cells (ADSCs) on PSCs in vitro.Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation.The coculture system of ADSCs and PSCs was set up by transwell insert.The proliferation of PSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of PSCs were tested by Western blot.The apoptosis of HSCs was tested by flow cytometer.The cytokines in the culture solution were tested by ELISA kit.Results The quantity of PSCs was above 5 × 106 cells per rat.The purity and the viability of the cells were about 90-97 percent.After coculture for 72 h,the proliferation and activation of PSCs was inhibited by ADSCs (F =223.27,P ＜ 0.05 ; F =52.97,P ＜ 0.05) and the apoptosis of PSCs was promoted by ADSCs (F =43.62,P ＜ 0.05).more NGF and less TGF-β1 was secreted by ADSCs than by PSCs (NGF:14.68 ± 0.94 vs.8.31 ±0.86,t =4.67,P ＜0.05;TGF-β1:10.65 ±0.46 vs.70.47 ±0.99,t =21.72,P＜0.01).Conclusions ADSCs inhibit the proliferation and activation of PSCs by ADSCs secreting cytokines.

Objective To study genotypes in nonsyndromic hearing loss (NSHL ) patients from Guangxi Zhuang Autonomous Region hearing speech rehabilitation center using DNA microarray in combination with Sanger sequencing .Methods Deaf patients received routine physical and otorhinolaryngoloical examinations as well as pure tone autiometry .Brainstem auditory evoked potential test was performed in uncooperative children .Blood samples were obtained from a total of 136 patients ,male 81 ,female 55 ,age from one year five month to seventeen ,having nonsyndromic hearing loss .Genomic DNA was extracted and then 9 hot mutation spots in 4 susceptibility genes were detected by DNA microarray .GJB2 and SLC26A was further detected by Sanger sequencing in the patients with negative results and heterozygotes .Results Among the 136 patients with nonsyndromic hearing loss ,20 cases were positive for GJB2 gene ,SLC26A4 gene or mitochondrial 12SrRNA gene mutations .There were 14 .71% (20/136)patients were positive for hot mutation spots in the deafness related genes ,25% (34/136)patients carried muta‐tions of deafness related genes using DNA microarray in combination with Sanger sequencing .Six SLC26A4 rare mutations (c .259G> T ,c .754C> T ,c .1229C> T ,c .1548_1549insC ,c .1705+5A>G and c .2086C> T) were de‐tected by Sanger sequencing .c .235delC was the most common mutation in GJB2 gene .c .919-2A>G ,c .754C> T and c .1229C> T were the common mutations in SLC26A4 gene .The mutation rate of GJB2 and SLC26A4 was 38 . 24% .and 58 .82% ,respectively .Conclusion Prevalent deafness-associated gene mutations in the nine loci studied were less frequently detected in nonsyndromic hearing loss patients from Guangxi Zhuang Autonomous Region hear‐ing speech rehabilitation center .It can improve the detection rate of deafness gene mutations by using gene microar‐ray in combination with Sanger sequencing .GJB2 and SLC26A4 are the common causative genes .

Objective:To analyze the clinicopathological characterization of primary esophageal benign tumor (EBT). Methods:A total of 1,058 EBTs were enrolled from 500,000 cases in an esophageal and cardiac tumor biological sample and clinical information data-base of Henan Key Laboratory for Esophageal Cancer Research (1973-2015) in the First Affiliated Hospital of Zhengzhou University. SPSS 21.0 software was applied for data analysis. Results:In this database, 1,058 cases with primary EBTs among the 249,246 esopha-geal tumor patients with detailed clinical and pathological information were identified with an incidence of 0.42%(1,058/249,246). A total of 544 patients were male with an average age of 50±11 years old, whereas 514 patients were female, with an average age of 52± 11 years old. Among the 10 types of EBTs, leiomyoma was the most common type (84.50%, 894/1,058), followed by papilloma (6.90%, 73/1058). Adenoma (0.38%, 4/1,058) was the rarest type. Leiomyoma, gastrointestinal stromal tumor, and neurofibroma mainly oc-curred in male patients. By contrast, lipoma, granulosa cell tumor, schwannoma, and hemangioma mainly occurred in female patients.All five cases of hamartoma occurred only in female patients. Given the incidence of≥50%as the common standard, the common EBT in sequence in young male patients was leiomyoma and gastrointestinal stromal tumor, whereas that in young female patients was granulosa cell tumor and lipoma. The common EBT in sequence in older male patients was papilloma, gastrointestinal stromal tumor, and leiomyoma, whereas that in older female patients was schwannoma, papilloma, leiomyoma, gastrointestinal stromal tumor, and hamartoma. Additionally, lipoma, hemangioma, neurofibroma, and adenoma in male patients and neurofibroma in female patients oc-curred in older patients. The different ages of patients with EBTs (P=0.034) and leiomyoma (P=0.004) had a statistical significance. In these EBTs, leiomyoma, papilloma, gastrointestinal stromal tumor, and schwannoma mainly occurred in the middle esophagus, where-as lipoma mainly occurred in the lower esophagus. The major treatment for EBT in the present study was surgery (57.54%, 492/855), which was followed by endoscopic resection (38.01%, 325/855) and others (4.45%, 38/855). Conclusion:The incidence of EBT is low, with a couple of different histological types. Gender, age, and predilection sites are different depending on the histological types of EBTs. Surgery and endoscopic resection are the major treatment methods.

OBJECTIVETo investigate the relationship between the molecular characteristics and phylogenetic evolution of rabies N gene.

METHODSSaliva samples were collected from rabies cases, and RT-PCR was used to amplify the N gene of rabies virus with the specific primers. The amplifying product of RT-PCR was cloned to pUCm-T vector and transformed into E.coli XL1-Blue and then the blue-white selection, PCR screening and gene sequencing were carried out to identify the positive clones. Finally, ExPASy and other bioinformatics software were used to analyze and predict the structure and biological characteristics of the N genome.

RESULTSThe amplification product of RT-PCR was 1 353 bp, the recombinant plasmid pUCm-T/N was constructed, the whole length of the N gene open reading frame was composed of 1 353 nucleotide residues to code 450 amino acids (20 kinds), the accession number submitted to the Genbank was HM756692, its sequence homology of nucleotides and amino acids compared with the vaccine strain CTN-1-V was 90% and 99% respectively. The evolutionary analysis showed that the isolated strain belonged to genotype I with certain geographic regionality.

CONCLUSIONThe characteristics investigation and bioinformatics analysis of Hunan0806 N gene will provide fundamental data to reveal the significance of the N gene characteristics for rabies epidemiology and its prevention & control.

Amino Acid Sequence ; Gene Expression Regulation, Viral ; physiology ; Humans ; Models, Molecular ; Molecular Sequence Data ; Nucleocapsid Proteins ; genetics ; metabolism ; Phylogeny ; Protein Conformation ; Rabies ; virology ; Rabies virus ; genetics ; metabolism ; Saliva ; virology

Objective To investigate the optimal thresholds of the passing rate with different gamma measurement criteria (percent dose difference/DTA) based on the Delta 4 three-dimensional dosimetric verification system in the verification of volumetric modulated arc-therapy (VMAT) plan for cervical cancer.Methods Thirty clinically-approved dual-arc VMAT plans using the RapidArcTM (Varian Medical Systems Inc.) for cervical cancer were randomly selected.The gamma analysis and dose-volume histogram (DVH) evaluation were performed using Delta 4.All the plans were classified according to the following two criteria:1.If the absolute percentage dose errors of all specific dosimetry indices on the DVH were less than 5％,the plan was regarded as clinically acceptable.2.If the gamma passing rate was 90％ or 95％ under the criteria of 2％/2 mm and 3％/3 mm,the plan was regarded as acceptable.The sensitivity and specificity analyses were conducted based on the classification results and the receiver operating characteristic (ROC) curve was plotted.By calculating the Youden Index,the optimal thresholds under different Gamma criteria (global and local 2％/2 mm and 3％/3 mm) were investigated.Finally,the ability of distinguishing the plan was clinically acceptable or not between the conventional and optimal thresholds was quantitatively compared according to the sensitivity and specificity analyses.Results The optimal thresholds under the global 3％/3 mm and 2％/2 mm criteria were 98.3％ and 87.05％;and 97.55％ 、86.05％ for the local gamma analysis.Compared with the conventional thresholds,the sensitivity of the optimal thresholds was 0.93 by using the global and local gamma analyses under the 3％/3 mm criterion.Under the 2％/2 mm criterion,the sensitivity of the optimal thresholds was 0.65 and the specificity was 0.49 by using the global gamma analysis.The sensitivity was 0.7 and the specificity was 0.46 by using the local gamma analysis,suggesting that the sensitivity and the specificity were more balanced under the 2％/2 mm criterion.Conclusions Application of the optimal thresholds in the verification of VMAT plans can maintain the balance between the sensitivity and specificity,prevent the harm of clinically unacceptable plans to patients to certain extent and reduce the probability of increasing the daily work load for physicists due to the misjudgement of clinically acceptable plans.

OBJECTIVE：To comp are the meridian tropism of compatible herbs and compatibility rules of ancient and modern prescriptions containing Eucommia ulmoides ，and to provide theoretical basis for modern clinical application of E. ulmoides . METHODS：Ancient prescriptions containing E. ulmoides were screened from Dictionary of TCM Prescription . The prescriptions containing E. ulmoides were screened from clinical trials which published in CNKI ，Wanfang database and VIP during Jan. 2000 to Jan. 2020. Data mining methods such as frequency statistics and Apriori algorithm were used to analyze the main diseases of ancient and modern prescriptions containing E. ulmoides and the use frequency ，types，meridian tropism and compatibility rules of medical materials. RESULTS ：A total of 84 ancient prescriptions and 188 modern prescriptions of E. ulmoides were included. Ancient prescriptions were mainly used to treat diseases such as back pain ，deficiency，threatened abortion. Modern prescriptions were mainly used to treat osteoarthropathy （mainly lumbar intervertebral disc herniation ），gynecological diseases ，cardiovascular diseases. Cinnamomum cassia ，Angelica sinensis and Achyranthes bidentatawerethe were the high-frequency compatible herbs in ancient prescriptions with E. ulmoides ，and were tonifying medicine ，dispelling wind and dampness medicine and clearing heat medicine；compatible herbs were mainly of warm property ，and the channel tropism was mostly kidney meridian and liver meridian；the five tastes of compatible herbs were mainly sweet and spicy. A. sinensis ，Glycyrrhiza uralensis and Rehmanniae Radix Praeparata were the high-frequency compatible herbs in modern prescriptions with E. ulmoides ，and were mainly tonic medicine，anti-rheumatic medicine ，medicine for promoting blood circulation and removing blood stasis ；compatible herbs were mainly of warm property ，and the channel tropism was mostly liver meridian and kidney meridian ；the five tastes of medicine were mainly bitter and spicy. By analyzing the Apriori association rules of high-frequency herbs ，a total of 10 core herbs combinations were obtained in ancient prescriptions ，including 8 third-order association rules and 2 fourth-order association rules ；a total of 11 core herbs combinations were obtained in modern prescriptions ，including 3 third-order association rules and 8 fourth-order association rules. CONCLUSIONS ： Ancient and modern prescriptions containing E. ulmoides are mainly used to treatlumbar disease ， followed by obstetrics and gynecology com diseases. In addition ，the modern prescriptions can also be #used for the treatment of cardiovascular and cerebrovascular diseases. The ancient prescriptions mainly focus on warming and tonifying ，while the modern prescriptions are more compatible with medicine for dispelling wind and dampness and medicine for promoting blood circulation and removing blood stasis on the basis of tonifying deficiency ；most of the medicines in ancient prescriptions contribute in kidney and liver meridians ，mainly sweet and pungent ，while most of the medicines in modern prescriptions contribute in liver meridian ，and are more compatible with bitter medicines.

Objective To investigate the optimal gamma passing rate of intensity-modulated radiotherapy (IMRT) dosimetric verification in the treatment of esophageal cancer using a three-dimensional dose verification system EDoseTM.Methods Twenty five esophageal cancer patients treated by 7-field IMRT were retrospectively reviewed.Measured dose distribution were reconstructed on CT image and evaluated by gamma analysis and DVH metrics using the EDoseTM system.Plans with DVH metrics dose difference ＜ 5％ or with gamma passing ＞ 90％ under 3％/3 mm criteria were accepted.The optimal gamma passing rate for criteria of 5％/3 mm,3％/3 mm,2％/2 mm were investigated by drawing the receiver operating characteristic (ROC) curves and calculating the Youden Index.The sensitivity and specificity of the these optimal thresholds in the plan verification were also analyzed.Results The optimal thresholds for global gamma indices with 5％/3 mm,3％/3 mm,2％/2 mm were 98.66％,94.84％,78.56％,respectively.In the 90％ common threshold,The sensitivity and specificity for common 90％ threshold and optimal threshold under 3％/3 mm criteria were 0.17 vs.0.85 and t 0.84 vs.0.27,respectively.The sensitivity and specificity were 0.89,0.65 and 0.23,0.47 for optimal thresholds under 5％/3 mm and 2％/2 mm criteria,respectively.Conclusions The sensitivity of optimal threshold gamma passing rate improved significantly compared with the common threshold (90％) at 3％/3 mm criteria.,The sensitivity and the specificity were more balanced at the 2％/2 mm criteria compared with those at 3％/3 mm criteria.

Objective:To investigate the application of 3D printing technique in percutaneous nephrolithotomy (PCNL) of patients with complicated kidney stones.Methods:From January 2015 to December 2017, 60 patients with complicated kidney stones were enrolled in the First Affiliated Hospital of Hebei North University for prospective study, and PCNL was proposed.All the patients were randomly divided into 3D print group (30 cases) and conventional image inspection group (30 cases, control group). Before operation, CT urography (CTU) was used in both groups.In 3D printing group, digital imaging and communications in medicine (DICOM) files of CT were extracted for 3D image postprocessing, and thermoplastic materials were used to print 3D model.In the 3D printing group, the digital imaging and communications in medicine (DICOM) files of CT were extracted for 3D image post-processing, and the 3D model was printed with thermoplastic materials.According to the comprehensive planning of 3D kidney model, a virtual safe and reliable percutaneous renal access was established for each patient, and PCNL was executed.The patients in the two groups were compared before, during and after operation.Preoperative: age, sex, body mass index, blood creatinine, stone size and CT value.During the operation: (1) the target renal calices location time; (2) the conformity between the preoperative planning and the actual operation; (3) the operation completion time.After operation: (1) stone removal rate; (2) hemoglobin reduction level; (3) postoperative recovery.Results:All the 60 patients successfully completed the operation, 30 patients successfully printed out the 3D model, which can accurately express the relationship between the stone and the adjacent anatomical structure, the internal renal artery and the collecting system.Positioning time of 3D printing group in target renal calices((2.9 ± 1.5) min vs.(5.8 ± 1.7) min, P=0.023), coincidence between simulated and actual puncture calices((89.5 ± 3.5)% vs.(60.2 ± 5.7)%, P=0.005), postoperative stone removal rate ((89.9 ± 4.5)% vs.(75.9 ± 5.2)%, P=0.009), and hemoglobin levels((1.4 ± 0.5) g/L vs.(2.9 ± 1.4) g/L, P=0.032) were superior to the control group, and the difference was statistically significant.But there was no significant difference between the two groups (all P>0.05). Conclusion:The 3D printed kidney model truly restores the anatomical details around the kidneys and stones, providing a stereoscopic and intuitive way to perform surgery, so it maybe has a significance guidance for percutaneous nephrolithotomy.

Trauma is the leading cause of death for people under 40 years old in the world. At present, the rescue and treatment system of trauma patients in China is not yet well established, and the mortality of trauma patients is higher than those in the developed countries. Improving the treatment system is the key to reducing the trauma mortality. In order to innovate the service mode of trauma first aid, further promote the establishment of regional trauma first aid system, improve the ability of trauma treatment, reduce the mortality and disability rate of trauma patients in Jiangxi Province, recently Health Commission of Jiangxi Province and the First Affiliated Hospital of Nanchang University have reached a consensus on the establishment of Jiangxi trauma first aid center. In order to provide reference for the construction of trauma treatment system, the author analyzes the following aspects including functional positioning, basic requirements, organization management, and evaluation of core indicators.

Objective: To assess clinical and radiographic outcomes of short implants (length six mm) in the posterior region and early-loading with splinted fixed dental prostheses. Methods: Forty-five patients were recruited at 3 centers and 2-3 implants (diameter 4 mm, length 6 mm) were implanted in each case. Totally, ninty-five implants were placed, using one-stage surgery protocol and loaded with a screw-retained splinted fixed prosthesis 6 weeks later. Clinical and radiographic examinations were performed preoperatively, post-surgery, at loading, and 6, 12 and 24 months after prosthesis placement. Results: Four implants were removed before loading. A total of 36 patients with 76 implants completed the whole treatment and 2 year follow-up according to the research design. No implant was lost after loading, and the 2-year survival rate of the implant was 100%(76/76). In all cases, the response of swelling and pain after operation was mild, and the bone resorption around the implants was not obvious at 6 months [(0.04±0.29) mm], 1 year [(-0.11±0.44) mm], and 2 years [(0.00±0.53) mm] after loading. Bone loss less than 1.00 mm was found only in 26% (18/69) implants after 2 years of loading, and the height of the alveolar bone around 17% (12/69) of the implant increased. During 2-year follow-up, the probing depth of the implants increased by nearly 0.50 mm, but there was no clinical significance (P>0.05). Conclusions The 2-year results of this study showed that the 6-mm-long implants can be a predictable procedure, especially in situations with limited bone height in posterior edentulous regions.