Objective:This study aims to investigate the determinants influencing the efficacy of microsurgical sperm retrieval in individuals diagnosed with idiopathic non-obstructive azoospermia (iNOA).Methods:A retrospective analysis was performed on the clinical data of 757 patients diagnosed with iNOA who underwent microsurgical sperm extraction at Peking University Third Hospital between January 2019 and December 2021. The median age of patients was 31(29, 33)years, and the duration of infertility was 3(2, 5)years. A total of 169 patients (22.3%) received preoperative pharmacological treatment with agents such as follicle-stimulating hormone, human chorionic gonadotropin, or aromatase inhibitors. Additionally, 327 patients (43.2%) underwent testicular biopsy (TESA) prior to surgery. Among these, 51 cases (15.9%) exhibited sperm presence on smear microscopy, while 57 cases (17.8%) demonstrated sperm presence on pathological examination. The pathological classifications of the biopsies included 102 cases (31.9%) of reduced spermatogenic function, 66 cases (20.6%) of delayed sperm maturation, and 63 cases (19.7%) of sertoli cell-only syndrome.Preoperative median pituitary prolactin(PRL)was 9.1(6.5, 12.5)ng/ml, follicle-stimulating hormone (FSH)20.1(14.2, 28.5)U/L, luteinizing hormone (LH)7.9(5.5, 11.3)U/L, testosterone(T)117 .0(81.3, 154.0)nmol/L, estradiol(E2)8.7(6.3, 11.8)pmol/L. Under general anesthesia, patients underwent microsurgical testicular incision for sperm retrieval.The surgical testicular volume was measured at a median of 6(5, 10) ml. Among the cases studied, 59 patients (7.7%) underwent left testicular surgery, 213 patients (28.1%) underwent right testicular surgery, and 485 patients (64.0%) underwent bilateral testicular surgery. Furthermore, 44 patients (5.8%) underwent a second microsurgical sperm retrieval procedure, while 4 patients (0.5%) underwent a third procedure.Based on the presence of sperm identified during the surgical procedure, participants were categorized into a sperm retrieval group and a non-sperm retrieval group. Clinical data of these two groups were analyzed. A subgroup analysis was performed on the observed indicators. Both univariate and multivariate logistic regression analyses were employed to examine the factors influencing the micro sperm retrieval rate.Results:Among the 757 iNOA patients, 255(33.7%) obtained sperm through micro sperm retrieval, while 502(66.3%) did not obtain sperm through micro sperm retrieval. The age of sperm-receiving group was higher than that of the non-sperm-receiving group [32(30, 35)years vs. 30(28, 33)years, P<0.01], and the course of infertility was longer than that of the non-sperm-receiving group [3.0(2.0, 5.5)years vs. 3.0(2.0, 4.0)years, P=0.004]. There was no significant difference in the sperm acquisition rate in the subgroup with or without preoperative drug treatment [38.5%(65/169)vs. 32.7%(185/566), P=0.164]. There was statistical significance in the sperm collection rate of different TESA results in subgroups [85.7%(24/28)of sperm were detected by microscopic smear and pathological examination and 75.9%(22/29)of sperm were detected by pathological examination and no sperm were detected by microscopic smear and 17% of sperm were not detected by microscopic smear and pathological examination (42/247), P<0.01). The rate of spermatogenesis in the subgroup with low spermatogenic function was significantly higher than that in the subgroup with spermatogenic maturation retardation and sercell-only syndrome [47.1%(48/112), 12.1%(8/66)vs. 11.1%(7/63), P<0.01]. There was no significant difference in testicular volume between the seminal and non-seminal groups [6.0(5.0, 10.0)ml vs. 6.0(5.0, 9.5)ml, P=0.862]. Pituitary prolactin [8.3(5.8, 12.0)ng/ml vs. 9.3(7.5, 13.0)ng/ml, P=0.001] and FSH[18.3(11.8, 27.4)U/L vs. 20.7(15.2, 28.7)U/L, P=0.005] in spermated group were lower than those in non-spermated group. Luteinizing hormone [7.6(5.1, 11.0)U/L vs. 8.0(5.6, 11.5)U/L, P=0.126], testosterone [8.8(6.0, 11.8)nmol/L vs. 8.7(6.4, 11.7)nmol/L, P=0.607], estradiol [124.0(87.8, 156.0)nmol/L vs. 114.5(79.9, 151.3)nmol/L, P=0.105] had no significant difference. The recovery rate of the first operation was higher than that of the second operation [97.7%(43/44)vs. 81.8%(36/42), P=0.032]. The sperm retrieval rate of bilateral operation was significantly lower than that of unilateral operation [6.0% bilateral (29/485)vs. 86.4% left (51/59)vs. 82.2% right (175/213), P<0.01]. The proportion of no sperm on one side of bilateral operation and only 4.7%(23/485)on the opposite side were obtained. The results of multivariate analysis showed that >30 and ≤40 years old subgroup ( OR=2.226, 95% CI 1.364-3.632, P=0.001), >40 and ≤50 years old subgroup ( OR=4.282, 95% CI 1.457-12.588, P=0.008)was higher than that of >20 and ≤30 years old subgroup. The sperm acquisition rate of the sperm subgroup was significantly increased by smear microscopy and pathological examination ( OR=6.486, 95% Cl 1.444-29.127, P=0.015), while the sperm acquisition rate of the sperm subgroup was not significantly decreased by smear microscopy and pathological examination ( OR=0.420, 95% Cl 0.200-0.881, P=0.022). The pathological type of puncture was associated with lower spermatogenesis maturation block ( OR=0.099, 95% CI 0.019-0.509, P=0.006). Higher FSH (>7.6 U/L)was associated with lower sperm yield ( OR=0.324, 95% CI 0.122-0.856, P=0.023). Conclusions:Age, FSH level, results of testicular biopsy and pathologic type of biopsy are independent factors affecting the sperm retrieval rate of iNOA patients undergoing micro-TESE. The success rate of sperm retrieval diminished following multiple surgical procedures. Furthermore, for patients who did not have sperm successfully retrieved from one side, that the likelihood of sperm retrieval from contralateral surgery would also be low.

Objective:This study aims to investigate the determinants influencing the efficacy of microsurgical sperm retrieval in individuals diagnosed with idiopathic non-obstructive azoospermia (iNOA).Methods:A retrospective analysis was performed on the clinical data of 757 patients diagnosed with iNOA who underwent microsurgical sperm extraction at Peking University Third Hospital between January 2019 and December 2021. The median age of patients was 31(29, 33)years, and the duration of infertility was 3(2, 5)years. A total of 169 patients (22.3%) received preoperative pharmacological treatment with agents such as follicle-stimulating hormone, human chorionic gonadotropin, or aromatase inhibitors. Additionally, 327 patients (43.2%) underwent testicular biopsy (TESA) prior to surgery. Among these, 51 cases (15.9%) exhibited sperm presence on smear microscopy, while 57 cases (17.8%) demonstrated sperm presence on pathological examination. The pathological classifications of the biopsies included 102 cases (31.9%) of reduced spermatogenic function, 66 cases (20.6%) of delayed sperm maturation, and 63 cases (19.7%) of sertoli cell-only syndrome.Preoperative median pituitary prolactin(PRL)was 9.1(6.5, 12.5)ng/ml, follicle-stimulating hormone (FSH)20.1(14.2, 28.5)U/L, luteinizing hormone (LH)7.9(5.5, 11.3)U/L, testosterone(T)117 .0(81.3, 154.0)nmol/L, estradiol(E2)8.7(6.3, 11.8)pmol/L. Under general anesthesia, patients underwent microsurgical testicular incision for sperm retrieval.The surgical testicular volume was measured at a median of 6(5, 10) ml. Among the cases studied, 59 patients (7.7%) underwent left testicular surgery, 213 patients (28.1%) underwent right testicular surgery, and 485 patients (64.0%) underwent bilateral testicular surgery. Furthermore, 44 patients (5.8%) underwent a second microsurgical sperm retrieval procedure, while 4 patients (0.5%) underwent a third procedure.Based on the presence of sperm identified during the surgical procedure, participants were categorized into a sperm retrieval group and a non-sperm retrieval group. Clinical data of these two groups were analyzed. A subgroup analysis was performed on the observed indicators. Both univariate and multivariate logistic regression analyses were employed to examine the factors influencing the micro sperm retrieval rate.Results:Among the 757 iNOA patients, 255(33.7%) obtained sperm through micro sperm retrieval, while 502(66.3%) did not obtain sperm through micro sperm retrieval. The age of sperm-receiving group was higher than that of the non-sperm-receiving group [32(30, 35)years vs. 30(28, 33)years, P<0.01], and the course of infertility was longer than that of the non-sperm-receiving group [3.0(2.0, 5.5)years vs. 3.0(2.0, 4.0)years, P=0.004]. There was no significant difference in the sperm acquisition rate in the subgroup with or without preoperative drug treatment [38.5%(65/169)vs. 32.7%(185/566), P=0.164]. There was statistical significance in the sperm collection rate of different TESA results in subgroups [85.7%(24/28)of sperm were detected by microscopic smear and pathological examination and 75.9%(22/29)of sperm were detected by pathological examination and no sperm were detected by microscopic smear and 17% of sperm were not detected by microscopic smear and pathological examination (42/247), P<0.01). The rate of spermatogenesis in the subgroup with low spermatogenic function was significantly higher than that in the subgroup with spermatogenic maturation retardation and sercell-only syndrome [47.1%(48/112), 12.1%(8/66)vs. 11.1%(7/63), P<0.01]. There was no significant difference in testicular volume between the seminal and non-seminal groups [6.0(5.0, 10.0)ml vs. 6.0(5.0, 9.5)ml, P=0.862]. Pituitary prolactin [8.3(5.8, 12.0)ng/ml vs. 9.3(7.5, 13.0)ng/ml, P=0.001] and FSH[18.3(11.8, 27.4)U/L vs. 20.7(15.2, 28.7)U/L, P=0.005] in spermated group were lower than those in non-spermated group. Luteinizing hormone [7.6(5.1, 11.0)U/L vs. 8.0(5.6, 11.5)U/L, P=0.126], testosterone [8.8(6.0, 11.8)nmol/L vs. 8.7(6.4, 11.7)nmol/L, P=0.607], estradiol [124.0(87.8, 156.0)nmol/L vs. 114.5(79.9, 151.3)nmol/L, P=0.105] had no significant difference. The recovery rate of the first operation was higher than that of the second operation [97.7%(43/44)vs. 81.8%(36/42), P=0.032]. The sperm retrieval rate of bilateral operation was significantly lower than that of unilateral operation [6.0% bilateral (29/485)vs. 86.4% left (51/59)vs. 82.2% right (175/213), P<0.01]. The proportion of no sperm on one side of bilateral operation and only 4.7%(23/485)on the opposite side were obtained. The results of multivariate analysis showed that >30 and ≤40 years old subgroup ( OR=2.226, 95% CI 1.364-3.632, P=0.001), >40 and ≤50 years old subgroup ( OR=4.282, 95% CI 1.457-12.588, P=0.008)was higher than that of >20 and ≤30 years old subgroup. The sperm acquisition rate of the sperm subgroup was significantly increased by smear microscopy and pathological examination ( OR=6.486, 95% Cl 1.444-29.127, P=0.015), while the sperm acquisition rate of the sperm subgroup was not significantly decreased by smear microscopy and pathological examination ( OR=0.420, 95% Cl 0.200-0.881, P=0.022). The pathological type of puncture was associated with lower spermatogenesis maturation block ( OR=0.099, 95% CI 0.019-0.509, P=0.006). Higher FSH (>7.6 U/L)was associated with lower sperm yield ( OR=0.324, 95% CI 0.122-0.856, P=0.023). Conclusions:Age, FSH level, results of testicular biopsy and pathologic type of biopsy are independent factors affecting the sperm retrieval rate of iNOA patients undergoing micro-TESE. The success rate of sperm retrieval diminished following multiple surgical procedures. Furthermore, for patients who did not have sperm successfully retrieved from one side, that the likelihood of sperm retrieval from contralateral surgery would also be low.

BACKGROUND: Sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) is a key protein that maintains myocardial Ca 2+ homeostasis. The present study aimed to investigate the mechanism underlying the SERCA2a-SUMOylation (small ubiquitin-like modifier) process after ischemia/reperfusion injury (I/RI) in vitro and in vivo . METHODS: Calcium transient and systolic/diastolic function of cardiomyocytes isolated from Serca2a knockout (KO) and wild-type mice with I/RI were compared. SUMO-relevant protein expression and localization were detected by quantitative real-time PCR (RT-qPCR), Western blotting, and immunofluorescence in vitro and in vivo . Serca2a-SUMOylation, infarct size, and cardiac function of Senp1 or Senp2 overexpressed/suppressed adenovirus infected cardiomyocytes, were detected by immunoprecipitation, triphenyltetrazolium chloride (TTC)-Evans blue staining, and echocardiography respectively. RESULTS: The results showed that the changes of Fura-2 fluorescence intensity and contraction amplitude of cardiomyocytes decreased in the I/RI groups and were further reduced in the Serca2a KO + I/RI groups. Senp1 and Senp2 messenger ribose nucleic acid (mRNA) and protein expression levels in vivo and in cardiomyocytes were highest at 6 h and declined at 12 h after I/RI. However, the highest levels in HL-1 cells were recorded at 12 h. Senp2 expression increased in the cytoplasm, unlike that of Senp1. Inhibition of Senp2 protein reversed the I/RI-induced Serca2a-SUMOylation decline, reduced the infarction area, and improved cardiac function, while inhibition of Senp1 protein could not restore the above indicators. CONCLUSION I/RI activated Senp1 and Senp2 protein expression, which promoted Serca2a-deSUMOylation, while inhibition of Senp2 expression reversed Serca2a-SUMOylation and improved cardiac function.
Animals ; Mice ; Calcium/metabolism* ; Cysteine Endopeptidases/metabolism* ; Myocardial Reperfusion Injury/metabolism* ; Myocardium/metabolism* ; Myocytes, Cardiac/metabolism* ; Proteins/metabolism* ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics*

Rats ; Animals ; Myocardial Reperfusion Injury/metabolism* ; Luteolin/metabolism* ; Down-Regulation ; Rats, Sprague-Dawley ; MicroRNAs/metabolism* ; Reperfusion Injury ; Apoptosis

Objective To investigate the effects of tongue cancer resistance-associated protein 1 (TCRP1) in proliferation of chronic myeloid leukemia cells (CML),and explore the new thoughts of pathogenesis of CML.Methods The expression of TCRP1 was detected in the peripheral blood mononuclear cells (PBMC) of CML with real-time quantitative polymerase chain reaction (PCR) and Western blot.After the expression of TCRP1 was interfered in K562 cells,the proliferation of cells was detected by 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and soft agar colony forming assay,and the expression of protein kinase B (AKT) and its phosphorylation were tested by Western blot.Results In PBMC of CML patients,the mRNA and protein levels of TCRP1 were significantly higher than those of normal controls.The results of MTS assay and soft agar colony forming assay showed that the proliferation of K562 cells was significantly decreased after the expression of TCRP1 was interfered.After knockdown of TCRP1 in K562 cells,the phosphorylation of AKT was significantly decreased while the expression of total AKT did not change.Conclusions The expression of TCRP1 was increased in CML cells.High expression of TCRP1 might contribute to proliferation of K562 cells via the phosphorylation of AKT.

AIM:To investigate the effects of Akt1 gene transfection into myocardium after ischemia-reperfusion (I/R) on mitochondrial permeability transition. METHODS:Forty adult male SD rats were divided randomly into five groups with 8 rats each:control group,I/R group,Ad-gene group,Ad-blank group and Ad-inhibitor group. The rats in Ad-gene group were injected with 30 μL Lipofectamine 2000 solution including Akt1 gene to the myocardium 48 h before ischemia while those in control group and I/R group were injected with PBS of the same volume. Rats in Ad-blank group were injected with Lipofectamine 2000 of the same volume into myocardium. In Ad-inhibitor group 30 μL Lipofectamine 2000 and gene complexes with LY294002 were injected. Hemodynamics,apoptotic index,the concentrations of lactate dehydrogenase,creatine kinase,the expression of Akt1,cytosolic,mitochondrial cytochrome C and MPT were also measured. RESULTS:The lowest level of Akt1 protein expression was observed in control group. The protein expression of Akt1 in Ad-gene group was higher than that in I/R group,Ad-blank group and Ad-inhibitor group. The AI,LDH and CK in Ad-gene group were significantly lower than those in other groups except control group. Transfection of Akt1 markedly reduced the loss of mitochondrial cytochome C after I/R injury. Ad-gene transfection led to a significant increase in absorbance at 540 nm compared to I/R group,Ad - black group and Ad-inhibitor group (P<0.05). CONCLUSION:Akt1 gene prevents myocardial apoptosis after I/R injury. Akt1 gene also inhibits the opening of mitochondria permeability transition and protects mitochondrial functions of myocardium in I/R injury.

Objective To compare the accuracy of real-time myocardial contrast echocardiography (RT-MCE) with DISA-SPECT in detecting hibernating myocardium. Methods Fourteen hospitalized patients with regional wall motion abnormalities (RWMA) underwent RT-MCE, DISA-SPECT, coronary angiography and percutaneous coronary intervention(PCI). Every left ventricular myocardial segment image was acquired and evaluated according 16-segment model of American Society of Echocardiography. The Motion of every segment was observed by echocardiography after 1,3,6 months. The gold standard is that the motion of ventricular segment get better after PCI. The images of RT-MCE and DISA-SPECT were analyzed semi-quantitatively. Results The sensitivity,specificity,accuracy of RT-MCE and DISA-SPECT in detecting hibernating myocardium were 74.60%, 91.7%, 81.3% and 93.3%, 75.0%, 86.2%, respectively. RT-MCE had more value in specificity while DISA-SPECT had more value in sensitivity. The correlation between two methods was good in detecting hibernating myocardium. Conclusions RT-MCE and DISA-SPECT can be used as valuable methods to detect hibernating myocardium, the correlation between two methods is good and they have more value in specificity and sensitivity respectively.

Objective To evaluate value of multi-indexes of real-time myocardial contrast echocardiography (RT-MCE) for the detection of coronary artery disease(CAD). Methods A total of 35 patients scheduled for coronary angiography underwent RT-MCE, and were undergone gated single photon emission computed tomography(gated-SPECT) after RT-MCE shortly. Coronary angiography was performed within one week of RT-MCE in all patients. All patients were divided CAD and no-CAD group. The observing indexes included: (1)The images of RT-MCE were analyzed quantitatively from microbubble replenishment curves for myocardial perfusion by using the Qlab software; A,β and A×β were compared between two groups. The sensitivity and specificity of RT-MCE for detection of CAD were performance by ROC curves Gensini score were calculated in 22 patients. Then the correlation analysis was used for comparing the value of A,β and A×β with Gensini score. (2) The sensitivity and specificity of gated-SPECT and RT-MCE for assessment of CAD were calculated by 4 score method. (3) RT-MCE for the detection of coronary artery stenosis with multi-indexes. Results (1) A,β and A×β were significant difference between two groups. The cutoff of A,β and A×β was 4. 58,0. 64 and 2. 73,then the sensitivity and specificity of RT-MCE for detection of CAD were 86. 0% , 80. 2% , 88. 9% and 84.1 % , 64. 6% , 79. 9 % , respectively. The correlation index was - 0. 79, - 0. 51 and - 0. 76 comparing the results of A, β and A ×β with Gensini. (2) The sensitivity and specificity of gated-SPECT for assessment of CAD were 84. 8 % and 82. 7% ,respectively. (3) The sensitivity of multi-indexes RT-MCE increased, the sensitivity was 89. l%,90. 4% and 96.3% when combinated A and β,A and A×β,and β and A×β, respectively. Conclusions RT-MCE with multi-indexes has a valuable application for assessment of CAD. The severity of CAD could be evaluated by RT-MCE.

Objective To investigate the effects of puerarin(Pur)on myocardial perfusion and ventricular wall motion in patients withacute coronary syndrome(ACS).Methods Thirty-seven patients with ACS were randomly divided into two groups:conventionaltreatment group(n=17,11 males,range of age:32-80 years,average age:60.9±4.9 years)and Purtreatment group(n=20,12 males,rangeof age:40-76 years.average age:62.7±3.5 years).Patients in the conventional treatment group received standard treatment according tothe current guidelines,while patients in the Pur treatment group received intravenous administration of Pur(500 mg/day)for 10 daysplus conventional treatment.Real-time myocardial contrast echocardiography (RT-MCE) was performed to evaluate the change inmyocardial perfusion index (MPI)and veiltricular wall motion index(VWMI)at admission and 10 days after treatment.Results At10 days after treatment,MPI was significantly higher(P＜0.01)and VWMI significantly lower(P＜0.01)in the Pur group comparingwith those in the conventional group.Conclusions Puerarin might improve myocardial microcirculation perfusion and ventricularwall motion in patients with ACS.

BACKGROUND: Acute myocardial ischemia can activate the c-Jun N-terminal protein kinase(JNK) and, in turn, the ischemia damage can be aggravated by JNK. While in chronic hibernating myocardium, whether chronic myocardial ischemia can activate JNK1/2 or not and what is the role of JNK1/2 in developing the chronic hibernating myocardium, is not clear.OBJECTIVE :To identify protein expression of JNK1/2 and the p-JNK1/2 changes in chronic hibernating myocardium.DESIGN: Randomly controlled experimental research.SETTING: This study was performed in Cardiovascular Disease Institute, the Affiliated Hospital of Xuzhou Medical College.MATERIALS: Animal models were prepared in the Catheter Department of the Affiliated Hospital of Xuzhou Medical College, protein expression of JNK1/2 and the phosphorylation(p-JNK1/2) changes were measured in the Department of Biochemistry of Xuzhou Medical College. Fourteen small Chinese domestic pigs were randomly divided into two groups, experiment group( n = 8) and control group( n = 6) . Right coronary artery was selected as target vessel in experiment group, and home-made constrictor was delivered through right femoral artery to induce chronic hibernating myocardium and myocardial infarction.MAIN OUTCOME MEASURES: Normal myocardial tissue in control group, normal myocardial tissue and chronic hibernating myocardium sample in experiment group were checked with light microscope and electron microscope,protein expression of JNK1/2 and the phosphorylation changes in myocardium tissue of three groups were analyzed by immunoblotting (Western blotting).RESULTS: p-JNK1/2 was higher in chronic hibernating myocardium of experiment group than that of normal myocardium in experiment and control group. Immunological activity of p-JNK1/2 in control group, normal myocardium, and chronic hibernating myocardium in experiment group was 1,1.42 ± 0.52, 2.6 ± 0.59, respectively.CONCLUSION: JNK1/2 can be activated in chronic hibernating myocardium tissue and also participated in its occurrence and development.