1.Expression,purification and bioimformatics analysis of allergen protein Profilin from silkworm (Bombyx mori)
Wei HU ; Zhilin LIANG ; Lianglu WANG ; Huiling ZHONG ; Zhigang LIU
Chinese Journal of Immunology 2017;33(1):81-84
Objective:To obtain recombinant Profilin of silkworm,identify its immunogenicity,predict its B cell epitopes and construct the evolutionary trees. Methods: The nucleotide sequence of Profilin was acquired from NCBI,synthesized it and cloned it into pET-28 vector. Then,the recombinant plasimids were transformed to E. coli BL21. After induced by IPTG,recombinant protein was purified by Affinity chromatography. Furtherly,its allergenicity was identified by Western blot,the potential B cell epitopes was analyzed through DNAStar and build the evolutionary trees by MEGA5. 05. Results: The recombinant protein of Profilin was successfully expressed and purified by affinity chromatography. Besides,the protein contains a high IgE-binding activity with IgE existing in serum of patients allergic to silkworm. Conclusion: The recombinant proflilin has IgE-binding activity, and it is meaningful for fundamental research and specific diagnosis studies of allergic diseases caused by silkworm.
2.Clinical significance of multi-tumor markers detection in the early diagnosis of hepatocellular carcinoma
Zengpeng LI ; Zhilin NIE ; Qinhong ZHANG ; Zhaoyang ZHONG ; Debing XIANG ; Feng JIN ; Ge WANG ; Dong WANG
Chinese Journal of Digestive Surgery 2009;8(2):113-115
Objective To investigate the value of combined detection of multi-tumor markers in the diagnosis of primary hepatocellular carcinoma (HCC) and to establish the discriminant equation. Methods Using a protein chip, 12 tumor markers in the serum from 98 patients with HCC and 67 patients with benign liver diseasewho had been admitted to Daping Hospital from November 2003 to April 2006, and 46 healthy individuals during he same period were analyzed. A discriminant equation was established to discriminate primary HCC from benign liver diseases. All the data were processed by variance analysis and chi-square test. Results The positive rates of the tumor markers were 89% (87/98) in patients with primary HCC, 19% (13/67) in patients with benign liver disease and 4% (2/46) in healthy individuals. There was statistical difference in the serum level of alpha-fetoprotein (AFP), eareinoembryonic antigen (CEA), ferritin (FER), CA19-9 and CA125 among the 3 groups (F =59.530, 40.472, 31.708, 75. 897, 153.066, P <0.05). Combined detection of AFP, CEA, FER, CA19-9 and CA125 improved the diagnostic accordance rate to 89%, which was significandy higher than the diagnostic accordance rate (64%) when only AFP was detected (X2 = 16.362, P <0.05). The accuracy of the discriminant equation was 90%. Conclusions Combined detection of multi-tumor markers is superior to AFP detection. Combined detection of multi-tumor markers can be used in screening of the HCC patients in HCC high risk population and in the early diagnosis of primary HCC.
3.The Application value of serum prostate specific antigen(PSA)in hyperplasia of prostate by YAG Laser
Xingxiang ZHONG ; Siping LIU ; Shiwu YAO ; Zhongkai SUN ; Liguo LIN ; Zhilin CHEN
Chinese Journal of Primary Medicine and Pharmacy 2012;19(5):654-656
Objective To explore the appliacation value of FPSAR between the serum f-PSA and t-PSA in hyperplasia of prostate by YAG Laser.Methods 150 cases with benign prostate hyperplasia(BPH)and 24 cases with prostate pcaancer were selected.The value of f-PSA、t-PSA、FPSAR was determined by TRFIA.Results The tPSA,f-PSA after treatment were significantly lower than before treatment in the two groups(t =2.984,2.701,P <0.05).The FPSAR after treatment was significantly higher than before treatment in the two groups(t =2.335,P <0.05);The patients of FPSAR≤0.05 in the overlapped field(t-PSA 4~45.5μg/L),the sensitivity of diagnosing PCa is 91.7%(22/24)[t-PSA > 45.5 μg/L(17/24)+(t-PSA 4~45.5 μg/L,FPSAR ≤ 0.15)(5/24)].To examine the PCa with high sensitiveity,it provided the reliable basis for selecting BPH correctly.The patients of BPH group after TUEP was followed up for 6~12 months.The t-PSA is(2.13 ± 0.45)μg/L、f-PSA is(0.85 ± 0.26)μg/L、FPSAR is (0.39 ± 0.06)μg/L.There is no significant difference compared with that after treatment for a month.The international prostate symptom score,(I-PSS)is from(28.3 ± 5.8)points dropped to(12.5 ± 2.1)points.The quality of life,(QOL)is from(4.1 ± 0.6)points dropped to(1.3 ± 0.1)points.The residual urine volume(RUV)is form(93 ±61)ml reduced to(15 ±9)ml.The urination after operation have improvedsignificantly.The Qmax is from average 6.3(2.6~9.5)ml/s before operation raise to 18.4(14.6~22.3)ml/s after operation.Campared with the pre-operation,there is significant difference.Conclusion The application of serum PSA was impoetantin case selection hyperplasia of prostate,comparison of the level changing before and after operation and following up the patients after operation by YAG Laser.
4.The diagnostic value of multi-slice CT on the congenital malformation of coronary sinus
Wei LI ; Xiaojing MA ; Qingjun SUN ; Xianfeng CHEN ; Yuan TAO ; Zhiyuan PENG ; Qingfeng XIONG ; Zhilin ZHONG ; Xin CHEN ; Yan CHEN ; Xuelian ZHANG ; Li WANG
Chinese Journal of Radiology 2012;46(2):110-112
Objective To investigate the application value of multi-slice spiral CT on the congenital malformation of coronary sinus. Methods MSCT finding of 98 patients with coronary sinus malformation confirmed by surgery were retrospectively analyzed,and the cases were divided into four categories based on the Mantini theory and comparison was made between the diagnosis from ultrasound and CT.A 2 × 2 table for Chi-square test was also used for statistics analysis.Results Among 98 patients,there were 72 patients with persistent left superior vena cava reflowed to right atria through coronary sinus,with 48 patients diagnosed by ultrasound and 72 patients by MSCT; there were 13 patients with anomalous pulmonary venous connection to coronary sinus,with 12 patients diagnosed by ultrasound and 13 patients by MSCT diagnosis; there were 10 patients with unroofed coronary sinus syndrome,with 6 patients diagnosed by ultrasound and 8 patients by MSCT,there were 2 patients with coronary sinus atresia,all diagnosed by MSCT; there were 1 patient with coronary sinus anomaly reflow to left arita.The significant difference between 2 modalities (x2 =22.7,P<0.01) shows that CT is superior to ultrasound.Conclusion MSCT is much more better than ultrasound in the diagnosis of malformation of coronary sinus and it can provide reliable diagnosis prior to surgery or interventional therapy.
5.Network pharmacology and molecular docking reveal the mechanism of resveratrol in oral squamous cell carcinoma treatment
CHEN Hongjun ; LEI Qi ; WANG Zhilin ; ZHONG Xiaowu ; QIU Ya ; LI Lihua
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(3):178-187
Objective:
To explore the molecular mechanism of resveratrol (RES) in the treatment of oral squamous cell carcinoma (OSCC) through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.
Methods:
The Swiss Target Prediction(http://www.swisstargetprediction.ch), SEA (http://sea.bkslab.org)database, and Pharm mapper database(http://lilab-ecust.cn) were used to retrieve RES-related targets, and the DISGENET (www.disgenet.org), OMIM (https://omim.org) and GeneCards (https://www.genecards.org) databases were used to screen OSCC disease targets. The intersection of drugs and disease targets was determined, and Cytoscape 3.7.2 software was used to construct a "drug-diseasetarget pathway" network. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database was used to construct a target protein interaction network, and the DAVID database was used for enrichment analysis of key proteins. Finally, molecular docking validation of key proteins was performed using AutoDock and PyMOL. The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC; western blot was used to determine the effect of resveratrol at different concentrations (50, 100) μmol/L on the expression of Src tyrosine kinase (SRC), epidermal growth factor receptor (EGFR), estrogen receptor gene 1 (ESR1), and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) signaling pathway proteins in OSCC HSC-3 cells.
Results:
A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified. A total of 116 potential common targets were obtained by intersecting drugs with disease targets. These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation, peptide tyrosine phosphorylation, transmembrane receptor protein tyrosine kinase signaling pathway, and positive regulation of RNA polymerase Ⅱ promoter transcription, and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects. The docking results of resveratrol with OSCC molecules indicated that key targets, such as EGFR, ESR1, and SRC, have good binding activity. The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC, EGFR, ESR1, p-PI3K, and p-AKT in HSC-3 cells in a dose-dependent manner.
Conclusion
RES can inhibit the expression of its targets EGFR, ESR1, SRC, p-PI3K, and p-AKT in OSCC cells.
6.Efficacy of Altemeier procedure in the treatment of rectal prolapse.
Yonglei CAO ; Yan ZHOU ; Congqing JIANG ; Guiyi YANG ; Hui SONG ; Lvfeng LIU ; Xu AI ; Jing ZHONG ; Zhilin GONG ; Jianhua DING ; Wei FU ; Qun QIAN
Chinese Journal of Gastrointestinal Surgery 2017;20(12):1370-1374
OBJECTIVETo evaluate the safety and efficacy of the perineal rectosigmoidectomy (Altemeier procedure) in the treatment of full thickness rectal prolapse.
METHODSClinical and follow-up data of 52 patients with full thickness rectal prolapse undergoing Altemeier procedure in 9 hospitals from September 2010 to July 2016 were analyzed retrospectively. Of which 38 cases were from Zhongnan Hospital of Wuhan University, 1 case from Suizhou Central Hospital of Hubei province, 1 case from Jingzhou Second People's Hospital of Hubei province, 3 cases from Wuxue First People's Hospital of Hubei province, 1 case from Jingmen First People's Hospital of Hubei province, 1 case from Tuanfeng County Hospital of Hubei province, 4 cases from Jingzhou Central Hospital of Hubei province, 2 from PLA Rocket Army General Hospital, 1 case from the Affiliated Hospital of Xuzhou Medical University in Jiangsu province. Altemeier operation steps: The line shaped teeth, the prolapsed rectum is first exposed to the anus. In the dentate line proximal 1-3 cm with ultrasonic knife or Ligasure ring outer rectal incision, using electric knife to mark pre resection line in rectal mucosa. Open down in front of the pelvic peritoneum. Incision of the outer intestine and the reduction of the internal rectum and part of sigmoid colon. To free and remove excess pelvic retroperitoneal, pelvic peritoneum and be at the top of the colon or rectum anterior pelvic reconstruction suture. The rear of the levator ani muscle forming rectum. Pull gently to the anus and rectum and sigmoid, in the absence of tension, 2-3 cm outside the anus was selected as the proximal inner bowel pre resection line, along the line of pre transection of proximal bowel resection, again the broken end of intestine full-thickness end-to-end anastomosis. Postoperative complication and recurrence were summarized. Gastrointestinal quality of life index (GIQLI), Wexner constipation score and Wexner fecal incontinence score were used to evaluate the efficacy.
RESULTSAll the 52 patients were beyond moderate full thickness rectal prolapse. Thirty-one were male and 21 were female with age ranging from 22 to 83 (average 53) years. The length of prolapsed rectum was 6 to 20 (average 9) cm and course of disease was 0.5 to 46(average 19.5) years. No perioperative death. Five patients (9.6%) had postoperative complications, including 2 anastomotic bleeding, 1 wall portion dehiscence of anastomosis, 1 anastomotic stenosis, and 1 malnutrition. Recurrence rate was 9.6%(5/52) within the long-term follow-up of 5 to 71 (median 40) years. Compared with the preoperative results, Wexner constipation score and Wexner fecal incontinence score decreased obviously (2.1±1.4 vs. 4.6±3.4, 4.8±4.1 vs. 6.8±4.1), and GIQLI significantly increased from 99.6±8.0 to 103.0±9.1 (all P<0.05) at 6-month after operation. Above 3 scores were sustained and continuously improved at 12-, 24-, and 36-month during the follow-up (all P<0.05).
CONCLUSIONAltemeier procedure possesses good efficacy with low morbidity of complication and recurrence in the treatment of full thickness rectal prolapse.