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Objective: To study the expression of transmembrane protein CMTM2 in the testis and sperm of adult males and to approach the potential function of the protein in the male reproductive system.Methods: The expression of CMTM2 in human testis and sperm was confirmed by Western blot.Immunohistochemical staining was used for detecting CMTM2 localization in the testis tissue, TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction, that is, immunofluorescent staining was used for detecting CMTM2 localization in both the testis and sperm before and after the acrosome reaction.Results: CMTM2 was presented in both human testis and sperm.In the testis, CMTM2 immunoreactive particles were observed mainly in the membrane of the different stages of spermatogenic cells.In the human sperm, its immunoreactivity was restrictively localized to the posterior head where sperm-egg fusion occurred, and the CMTM2 localization was not affected by sperm acrosome reaction.CMTM2 was widely expressed in seminiferous tubules of the human testis, mainly in the cell membranes of spermatogenic cells, which was consistent with the previous reports.The immunofluorescence performed on frozen human testis slides showed similar findings with immunohistochemistry, which gave weight to the localization of CMTM2 in the cell membranes of spermatogenic cells at different stages.TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction.CMTM2 was localized at the posterior head of sperm before and after acrosome reaction.The localization and expression of CMTM2 were not affected by sperm acrosome reaction.Conclusion: Expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.The expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.However, it still remains to be further elucidated about the definite role of CMTM2 in male reproductive system and the process of spermatogenesis.And in vitro fertilization experiments are needed to confirm the role of CMTM2 in fertilization in future.
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Objective To evaluate the health-related quality of life (HRQOL) for prostate cancer patients receiving androgen deprivation therapy (ADT).Methods We studied 200 patients with prostate cancer who were treated with primary ADT in Peking University People's Hospital from June 2014 to June 2015.The patients'average age was 73.9 years.The mean PSA level was 21.57 ng/ml when they were diagnosed with prostate cancer.Of these 200 patients,79％ (158/200) were localized and seclected ADT therapy due to age,body condition,basic diseases or individual will.21％ (42/200) were locally advanced or metastasic,which accord with the indication of ADT therapy.The scales,including the MOS item short from health survey (SF-36),Functional Assessment of Cancer Therapy-General module (FACT-G),Functional Assessment of Cancer Therapy-Prostate instrument (FACT-P),Self-Rating Depression Scale (SDS) and Self-Rating Anxiety Scale (SAS) were used to evaluate their HRQOL.Results According to results of SF36,the physical health component score and mental health component score was 67.41 ± 16.39 and 64.81 ± 17.51,respectively.They indicated that the overall quality of life of these patients was at an acceptable level.And the average score of all domains exceeded 60 except general health domain,which the score was only 40.03 ± 21.89.When it comes to FACT-P,the sum score,emotional well-being score and prostate cancer component score were 8.15 ± 3.72,12.30 ± 4.04 and 77.41 ± 9.95,respectively,which were less than half of their respective top score.However the physical well-being score was 20.41 ±4.29,which was a relatively satisfactory value.In addition,61％ (121/200) patient selected not at all in item I am able to have and keep an erection.The score of SDS and SAS was 46.76 ± 8.29 and 43.25 ±9.69,respectively.And there were 23.0％ (46/200) and 21.5％ (43/200) patiens exhibited depression and anxiety,respectively.Conclusion In the present study,the prostate cancer patients receiving ADT showed acceptable HRQOL,but some patients sufferd from depression,anxiety and erectile dysfunction.
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Objective:To investigate whether chemokine like factor (CKLF)-like myelin and lympho-cyte and related proteins for vesicle trafficking and membrane link (MARVEL)transmembrane domain-containing protein 2 (CMTM2)is involved in spermatogenesis in varicocele induced sub-fertility rats and to discuss the possible mechanisms.Methods:Forty male SD rats (body weight:220 -330 g,age:6 -7 weeks)were randomly divided into 4 groups:varicocele for 4 weeks,varicocele for 12 weeks,sham operation for 4 weeks and sham operation for 12 weeks,with 10 rats in each group.These rats were intro-duced by partially ligating left kidney veins for the experimental groups,and the sham surgery groups as controls were executed with exactly the same surgery as in the experimental groups except for the ligation. The rats in control and experimental groups for 4 and 12 weeks were killed after laparotomy at the end of 4 and 12 weeks,respectively,the left testes and epididymis were taken out for counting the sperm,ob-serving the seminiferous tubule change and immunochemistry for CMTM2.The changes included sperm density and motility,the outer diameter and inner diameter change and the changes of epithelium and the CMTM2 expression in immunochemistry.Results:Compared with the control groups,the sperm density [(63.9 ±7.1)×106 /mL vs.(74.3 ±5.0)×106 /mL]and motility[(58.7% ±7.9%)vs.(66.1% ± 4.3%)]were reduced slightly in group of varicoele for 4 weeks,respectively (t =1.432,1.563;P =0.076,0.059,respectively ).Varicocele significantly caused a decrease in sperm concentration [(40.5 ±7.2)×106 /mL vs.(71.1 ±4.5)×106 /mL]and motility [(35.2% ±8.5%)vs.(63.4% ± 4.1%)]at 12 weeks,compared with the related sham groups (t =3.754,3.933;P =0.004,0.002, respectively).Additionally,testis CMTM2 exhibited the same disparity,that is,the CMTM2 protein ex-pression in varicocele group was significantly reduced,with the ratio of sham group to varicocele group at the end of 12 weeks 2.3 ±0.4 (t =1.978;P =0.039).In the evaluation of seminiferous tubules diame-ter,the external [(198.2 ±10.2)μm vs.(255.8 ±12.7)μm,t =2.125,P =0.003]and epithelium diameter [(54.1 ±1.5)μm vs.(75.5 ±4.1)μm,t =2.246,P =0.021]were decreased compared with the sham-related groups and previous varicocele groups.In all the varicocele groups,all types of sperm motility decreased compared with the related sham-operated group (P <0.05).Conclusion:This study suggests varicocele has a detrimental effect on CMTM2 levels and decreases spermatogonia cell number,seminiferous tubules diameter,and sperm indices.CMTM2 is associated with sperm changes in rats with varicocele,and further studies are needed to study the mechanism.
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Objective:To investigate the expression of MEK/ERK signaling pathways in renal cell car-cinoma with bone metastasis,and to analyze the differences of expressions of VEGFR-2,MEK,ERK on the primary and metastasis tissue and its mechanism.Methods:The tissue samples were obtained from 7 renal cell carcinoma patients kindly provided by Department of Urology,Peking University People’s Hos-pital from January 1,2009 to January 1,2010.The expression of MEK/ERK signaling pathways was de-tected in the 7 renal cell carcinoma patients`primary and matched metastatic tissues with ICH,The anti-body concentrations were 1 ∶200,1 ∶25,and 1 ∶250,respectively.The mutation of the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the se-cond exon of the MEK1 gene were detected with PCR.Results:The expression intensities of VEGFR-2, MEK,and ERK were measured by H-score [intensity (1,2,3,or 4)multiplied by the distribution (%)].VEGFR-2,MEK,and ERK expressions were divided into 3 groups according to the positive dis-tribution of the tumor cells:1,0 -5%;2,6% -50%;and 3,>50%,To assess intratumor heteroge-neity,three distinct microscopic fields (×200)from each specimen were used to evaluate the expres-sions,Subsequently,the scores were averaged to obtain a single concatenated score for each tissue. VEGFR-2,MEK,and ERK expressions were assessed by 2 independent pathologists who were blinded to the clinicopathological data.The data were expressed as the mean value of the triplicate experiments.The expressions of MEK,and ERK were higher in the metastatic tissues than in the matched RCC tissues (6.10 ±4.10 vs.1.33 ±0.51,P =0.015;9.10 ±2.24 vs.4.43 ±2.84,P =0.021 )while the ex-pression of VEGFR-2 was not different between the primary and metastatic tissues (P =0.901).No mu-tation was detected on the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the second exon of the MEK1 gene.Conclusion:MEK/ERK signa-ling pathways may play an important role in the metastasis and the resistance of sunitinib in RCC patients with bone metastasis.
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Objective:To investigate the change of biological characteristics after stable knockdown of CKLF-like MARVEL transmembrane domain containing 3 (CMTM3)expression in PC3 by lentivirus shRNA and to reveal new therapeutic targets.Methods:The research includes two groups:sh393 is the experimental group in which CMTM3 is knocked down in PC3 cell line;shN is the control group in which CMTM3 is negatively knocked down.The expression of CMTM3 was detected by Western blot.The mi-gration ability of PC3 after stable knockdown was detected by Transwell and Wound healing assay.The invasion ability of PC3 was detected by Matrigel assay.Results were obtained from at least three indivi-dual experiments.Results:The expression of CMTM3 in sh393 group is significant lower than shN group (0.004 0 ±0.000 4 vs.0.490 0 ±0.055 7,P <0.001)detected by Western blot.It also had statistical significance in Matrigel assays (248.6 ±4.5 vs.113.0 ±3.3),Transwell (203.6 ±1.9 vs.103.0 ± 1.2)and Wound healing assays (95.0 ±2.9 vs.33.0 ±1.5)that knockdown of CMTM3 promoted mi-gration,and invasion of PC3 cells in vitro (P <0.001).Conclusion:Negative correlation exists between the stable knockdown of CMTM3 and change of biological characteristics in PC3 cells,and knocking down CMTM3 affects migration,and invasion ability in PC3 cells.