1.Polymorphism and Tissue Expression Analysis of TYR and MC1R Genes in Guinea Pigs with Different Coat-Color Phenotypes
Yingen TANG ; Yaxian FENG ; Min ZHONG ; Zhen WEI ; Lie WANG ; Diwen LIU
Laboratory Animal and Comparative Medicine 2025;45(1):21-29
Objective To explore the polymorphism of tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes and their mRNA expression levels in relation to coat-color phenotypes in guinea pigs, providing genetic markers for locating dominant traits in guinea pigs. Methods A total of 57 self-bred ordinary-level guinea pigs were selected and divided into three groups based on coat color: white (n=22), variegated (n=22) and black (n=13). The guinea pigs were euthanized with an overdose of pentobarbital sodium via intraperitoneal injection. DNA was then extracted from the dorsal skin tissue. Polymorphism in the coding sequence (CDS) of the exons of the TYR and MC1R genes in each group was detected by cloning and sequencing. The mRNA expression of the two genes in skin tissues was detected by real-time fluorescent quantitative PCR to investigate the relationship between these genes and guinea pig coat color. Results A single nucleotide polymorphism (SNP) site was found in the CDS region of TYR exon Ⅰ, where the base A was replaced by G. All white guinea pigs had the G/G genotype for TYR, while no deep-colored (variegated and black) guinea pigs exhibited the G/G genotype for TYR. Most deep-colored guinea pigs had the A/A genotype, and a few had A/G genotype. The A/A genotype frequency in black guinea pigs was higher than in variegated guinea pigs. A 2 760 bp sequence deletion was identified in the exon of the MC1R gene, marked as the - gene, with non-deleted samples marked as N gene. Most white guinea pigs had the -/- genotype for MC1R, variegated guinea pigs mainly had the -/N genotype, and black guinea pigs mainly had the N/N genotype, with a few showing the -/N. The TYR gene expression level was higher in white guinea pigs, lower in variegated guinea pigs, and intermediate in black guinea pigs, but there was no significant difference among the three groups (P>0.05). The MC1R gene expression level in white guinea pigs was extremely low, while both variegated and black guinea pigs showed significantly higher levels than white guinea pigs (P<0.01). Black guinea pigs showed significantly higher levels than variegated guinea pigs (P<0.05). ConclusionThe TYR and MC1R genes synergistically regulate coat color of guinea pigs. The G-site mutation in the TYR gene may lead to albinism, and the change of N-site in the MC1R gene affects the depth of the coat color.
2.Study progress in erectile dysfunction after radical pelvic surgery
Ruiyu LI ; Qiang FU ; Aiyun ZHU ; Keqin ZHANG ; Dingqi SUN ; Tongxiang DIAO ; Shuai LIU
Chinese Journal of Urology 2023;44(1):75-78
Radical pelvic surgery is a surgical method mainly used to treat tumors in the pelvic cavity, and erectile dysfunction (ED) is a common sexual dysfunction after surgery. The incidence of ED after radical pelvic surgery is not uniformly understood due to differences in surgical approaches and methods of investigation. The main causes of postoperative ED include intraoperative neurovascular injury, psychosomatic factors and preoperative patient characteristics, with intraoperative injury to the neurovascular bundle being the most common. Studies have shown that the occurrence of postoperative ED can be prevented by active intervention, but still lack of effective treatment measures. This article reviews and summarizes the clinical epidemiological features and research progress in recent years on ED after radical pelvic surgery, and discusses specific measures for the prevention and treatment of postoperative ED.
3.Effects of maximal androgen blockade therapy on serum calcium, phosphorus and other metabolic indices in elderly patients with prostate cancer
Shicong LAI ; Xuan WANG ; Tongxiang DIAO ; Shengjie LIU ; Xingbo LONG ; Zijian TIAN ; Jianyong LIU ; Huimin HOU ; Jianye WANG ; Ming LIU
Chinese Journal of Geriatrics 2020;39(11):1331-1334
Objective:To investigate the effect of maximal androgen blockade(MAB)therapy on serum calcium, phosphorus and other metabolic indices in elderly patients with prostate cancer.Methods:Clinicopathological data of prostate cancer patients treated with MAB in our department from January 2010 to December 2018 were retrospectively analyzed.All patients underwent prostate biopsy for definitive diagnosis.Detailed data on patient's age, body mass index(BMI), previous medical history, treatment plan and peripheral blood indicators before and after endocrine treatment, such as blood calcium, phosphorus, hemoglobin, fasting blood glucose, triglycerides and cholesterol, were collected.Results:Patients had a mean age of(75.5±5.8)years and a mean BMI of(24.6±3.2)kg/m 2.Blood calcium levels exhibited a downward trend after MAB treatment compared pre-treatment[(2.12±0.44)mmol/L vs.(2.17±0.31)mmol/L, t=0.82, P=0.42], but had no significant difference.Serum phosphorus concentrations were higher and the calcium-phosphorus ratio was lower after MAB treatment than before treatment[(1.02±0.26)mmol/L vs.(1.17±0.34)mmol/L, 2.10±0.28 vs.1.88±0.60, t=-4.12 and 3.56, P<0.01]. After MAB treatment, blood fasting glucose[(6.50±1.55)mmol/L vs.(5.34±1.04)mmol/L, t=-7.82, P<0.01], triglycerides[(1.66±1.32)mmol/L vs.(1.22±0.59)mmol/L, t=-3.38, P<0.01]and cholesterol[(4.70±1.08)mmol/L vs.(4.16±0.90)mmol/L, t=-4.72, P<0.01]were elevated, while hemoglobin concentrations[(122.11±20.43)g/L vs.(130.78±23.98)g/L, t=3.98, P<0.01]were decreased compared with pre-treatment levels. Conclusions:MAB therapy can cause varying degrees of metabolic abnormalities in calcium and phosphorus metabolism, hemoglobin concentrations, blood glucose and lipid levels in elderly prostate cancer patients.The above indicators should be closely monitored during treatment, and treatment-related complications should be proactively prevented.
4.Interaction between ACOT7 and LncRNA NMRAL2P via Methylation Regulates Gastric Cancer Progression
Yonsei Medical Journal 2020;61(6):471-481
Purpose:
Gastric cancer (GC) has a very poor prognosis when diagnosed at a late stage. Acyl-CoA thioesterase 7 (ACOT7) is a major isoform of the acyl coenzyme family that catalyzes the hydrolysis of fatty acyl-CoAs into unesterified free fatty acid and coenzyme A. The purpose of this study was to investigate the expression levels of ACOT7 in GC and mechanisms related therewith.
Materials and Methods:
Screening of systematic biology studies revealed ACOT7 as a key gene in GC, as well as involvement of the long non-coding RNA NMRAL2P in ACOT7 expression. In this study, GC tissues and adjacent tissue samples were obtained from 10 GC patients at the Department of Gastrointestinal Surgery. GES1 and SGC-7901 cells were collected and treated to silence ACOT7 and overexpress NMRAL2P. The expressions of ACOT7 and NMRAL2P were detected by real-time quantitative PCR and Western blot. Additionally, cell proliferation, apoptosis, migration, and invasion were examined.
Results:
ACOT7 was upregulated in gastric tumor tissues and GC cell lines. ACOT7 gene silencing induced a less malignant phenotype and was closely correlated to reduced cell proliferation and migration, altered cell cycle, and increased apoptosis. Furthermore, NMRAL2P was downregulated in tumor tissues and GC cell lines. NMRAL2P overexpression induced a more malignant phenotype and significantly inhibited the expression of ACOT7. Importantly, NMRAL2P indirectly methylated ACOT7 by binding to DNMT3b, thereby suppressing ACOT7 expression.
Conclusion
NMRAL2P activation suppresses ACOT7 expression in GC. Thus, ACOT7 could be a promising target for the treatment of GC.
5.Practice of PDCA Method in the New Specialties Accreditation of Drug Clinical Trial Institution
Liping MA ; Xiangming JIANG ; Bing LIU ; Tongxiang ZHAO ; Hongyu WANG ; Pingchao XIANG ; Haiying WANG
China Pharmacy 2018;29(7):869-872
OBJECTIVE:To explore the effects of PDCA method in the new specialties accreditation of drug clinical trial institution. METHODS:PDCA method was used for drug clinical trial institution office and 9 newly applied majors in our hospital. Total score of each major were compared before and after intervention,in order to make the newly applied major meet the standard of specialties accreditation of drug clinical trial. RESULTS:After conducting PDCA related training,establishing new specialties accreditation work group,formulating work goals and plans,9 new majors of our hospital were all approved by CFDA for new specialties accreditation;after intervention total score of each accreditation item for newly applied major were all higher than before intervention,with statistical significance(P<0.05),improved by more than 45.57%. CONCLUSIONS:It is feasible to adopt PDCA method in the preparation of new specialty accreditation of drug clinical trials. It is of significance to guarantee scientific and reliable drug clinical trial results and protect the rights and interests of the subjects.
6.The characteristics and independent associated factors of positive surgical margin after radical prostatectomy
Huimin HOU ; Xin JIANG ; Ming LIU ; Tongxiang DIAO ; Jianye WANG
Chinese Journal of Urology 2018;39(10):740-744
Objective To analyze the associated factors of positive surgical margin after radical prostatectomy.Methods A retrospective analysis was conducted on 320 patients who underwent radical prostatectomy from June 2007 to June 2017,whose age was 45-80 years(mean 67.9) and PSA was 0.05-123.19 ng/ml (mean 14.4ng/ml).The patients were divided into groups by age,PSA,PI-RADS score,clinical stage,biopsy Gleason score and operation approach.Chi-square test was used for single factor analysis and binary logistic regression analysis for multivariate analysis to evaluate the correlation between clinical and pathological data and positive cutting edge.Result Among the total 320 patients,there were 94 (29.4%) patients had positive surgical margin after radical prostatectomy.There were 26 (21.0%) positive surgical margin located at ventral sites,18(14.5%) located at dorsal sites,21 (16.9%) located at base,and 59(47.6%) located at tip.The positive rate of surgical margin was different in different positive areas of MRI (P <0.01),among which the MRI showed cancer located in the tip of prostate had the highest positive rate (47.6%) of surgical margin after prostatectomy.Univariate risk factor analysis was performed which showed that PSA(P =0.023),positive needle percentage (P =0.001),biopsy pathologic Gleason score(P =0.029),PI-RADS score (P =0.022) and prostate cancer risk score (P =0.006) had significant correlation with positive surgical margin.The age (P =0.257),clinical stage (P =0.161) and operation approch (P =0.260) had no significant correlation.Then multivariate analysis showed that PI-RADS score (P =0.023) and positive needle percentage (P =0.047) could be used as independent predictors of positive surgical margin.Conclusions PI-RADS score and percentage of positive biopsy needles were independent risk factors for positive surgical margin after prostatectomy.There was highest positive rate of surgical margin when MRI showed cancer located at the tip of prostate.
7.Effects of total flavonoids from Cycas Revolute on expression of VEGF,bFGF,HIF-1α and NF-κB in model mice of Lewis lung cancer
Shaohui WANG ; Sibu MA ; Yu YAN ; Zhekang CHENG ; Yachen ZHANG ; Tongxiang LIU
Chinese Journal of Immunology 2017;33(7):1029-1034
Objective:To investigate the effects of total flavonoids from Cycas Revolute on expression of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF),Hypoxia inducible factor-1α (HIF-1α) and nuclear factor-κB(NF-κB) in model mice of Lewis lung cancer.Methods: The expressions of VEGF,bFGF,HIF-1α and NF-κB in tumor tissues were detected by immunohistochemistry and Western blot.The expression of VEGF,VEGF and NF-κB in tumor tissues were detected by fluorescent quantitative PCR.BFGF,HIF-1α and NF-κB mRNA were detected by immunohistochemistry.Results: The results of immunohis-tochemistry,Western blot and Real-time PCR showed that the results were basically the same,compared with model group,the expression of VEGF,bFGF,HIF-1α,NF-κB mRNA and the expression of VEGF,bFGF,HIF-1α and NF-κB were decreased,the difference was highly significant (P<0.05).Conclusion: The mechanism of total flavonoids from Cycas Revolute in the treatment of lung cancer may be through inhibition of the expression of VEGF,bFGF,HIF-1α,NF-κB in invasion and metastasis,and further inhibit the expression of VEGF,bFGF,HIF-1α and NF-κB in invasion and metastasis-related proteins,thus play a role of anti-lung cancer invasion and metastasis.
8.Optimization of Ultrasonic Extraction of Total Saponins from Marsdenia tenacissima by Box-Behnken Re-sponse Surface Method and Its Purification Technology
Dongwei XU ; Shaohui WANG ; Tongxiang LIU
China Pharmacy 2016;27(25):3535-3538
OBJECTIVE:To optimize the ultrasonic extraction and purification technology of total saponins from Marsdenia te-nacissima,and to prepare purified total saponins from M. tenacissima. METHODS:Using the extraction rate of total saponins from M. tenacissima as index,ethanol volume fraction,solvent-solid ratio and extraction temperature as factors,ultrasonic extraction technology of total saponins from M. tenacissima was optimized by single factor test and Box-Behnken response surface design. Us-ing the content of total saponins in elution,the effects of ethanol elution volume fraction on purification technology of total sapo-nins by D101 type macroporous adsorption resin was investigated. RESULTS:The optimal ultrasonic extraction technology was as follows as ethanol volume fraction of 85%,solvent-solid ratio of 18.5∶1 (ml/g),extraction temperature of 80 ℃. In verification test,the average extraction rate of total saponins from M. tenacissima was 4.80%(RSD=1.06%,n=3);it was close to predicted value 4.89%,and the deviation was 1.84%. The optimal purification technology was as follows as 45% ethanol as eluant,90%ethanol for purification. Verification test showed that the average content of total saponins from M. tenacissima was 62.45%(RSD=0.88%,n=3). CONCLUSIONS:The optimized technology can be used for the extraction and purification of total saponins from M. tenacissima. The technology is stable and reliable.
9.Research Progress on Chemical Constituents, Pharmacological Effects and In-vivo Metabolism of Gynostemma Pentaphyllum
Shaohui WANG ; Daojin CHEN ; Tongxiang LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(11):2389-2393
Gynostemma pentaphyllum belonged to the Cucurbitaceae family.It was also called theQi-Ye-Dan,licorice stem,Gong-Luo Guo-Di,and etc.It contained saponin,flavonoids,polysaccharides,terpenoids and other chemical compositions.Its pharmacological effects included cholesterol-lowering activity,antitumor activity,hypoglycemic activity,anti-aging and boost immunity.This paper reviewed the research progress on chemical constituents,pharmacological effects and in vivometabolism of gynostemma pentaphyllum,in order to provide a theoretical basis for its further research and development.
10.Anti-tumor Effect of Tenacissoside H and Influence on Immune Function of Lewis Lung Cancer Mice
Wenjing WEI ; Guanqing ZHANG ; Shijie LIU ; Tongxiang LIU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(4):63-66
Objective To study the anti-tumor effect of tenacissoside H on Lewis lung cancer mice, and explore its impacts on immune function of tumor-bearing mice. Methods Mice was injected Lewis lung cancer cells subcutaneously to the right axilla. Thirty successful tumor-bearing mice were randomly divided into model group, the cyclophosphamide (CTX) group and tenacissoside H group, 10 mice in each group. Three days after inoculation, mice were intraperitoneally injected by normal saline, CTX and tenacissoside H respectively every two days, 0.2 mL in each mouse. On the 21st day, the eyeballs were extracted and blood was drawn, tumor tissue, spleen and thymus were taken and weighted to calculate tumor inhibition rate, spleen index and thymus index, and the contents of IL-2 and IL-10 were detected by ELISA. Results The tumor weights of CTX group and tenacissoside H group were lower than that of the model group with significant difference (P<0.05), and the tumor inhibition rates were 54.12%, 25.68%. The thymus index and spleen index of tenacissoside H group increased, but that of CTX group decreased significantly. Compared with the model group, the IL-2 level of tenacissoside H group was significantly increased, while the IL-10 level decreased. Conclusion Tenacissoside H can inhibit growth and metastasis of Lewis lung cancer, regulate the expression of IL-2 and IL-10, and improve the immune function of tumor-bearing mice.

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