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Jianbin Guo,1,2,* Guihua Cao,3,* Xing Wang,1 Wenhao Tang,1 Weilong Diwu,1 Ming Yan,1 Min Yang,1 Long Bi,1 Yisheng Han1 1Department of Orthopedics, The First Affiliated Hospital of Air Force Military Medical University, Xi’an, People’s Republic of China; 2Department of Joint Surgery, Hong-Hui Hospital, Xi’an Jiaotong University College of Medicine, Xi’an, People’s Republic of China; 3Department of Geriatrics, The First Affiliated Hospital of Air Force Military Medical University, Xi’an, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yisheng Han; Long Bi Email drhanys@fmmu.edu.cn; bilong@fmmu.edu.cnIntroduction: With increases in implant infections, the search for antibacterial and biofilm coatings has become a new interest for orthopaedists and dentists. In recent years, graphene oxide (GO) has been extensively studied for its superior antibacterial properties. However, most of these studies have focused on solutions and there are few antibacterial studies on metal surfaces, especially the surfaces of cobalt-chromium-molybdenum (CoCrMo) alloys. ϵ-Poly-L-lysine (ϵ-PLL), as a novel food preservative, has a spectrum of antimicrobial activity; however, its antimicrobial activity after coating an implant surface is not clear.Methods: In this study, for the first time, a two-step electrodeposition method was used to coat GO and ϵ-PLL on the surface of a CoCrMo alloy. Its antibacterial and antibiofilm properties against S. aureus and E. coli were then studied.Results: The results show that the formation of bacteria and biofilms on the coating surface was significantly inhibited, GO and ϵ-PLL composite coatings had the best antibacterial and antibiofilm effects, followed by ϵ-PLL and GO coatings. In terms of classification, the coatings are anti-adhesive and contact-killing/inhibitory surfaces. In addition to oxidative stress, physical damage to GO and electrostatic osmosis of ϵ-PLL are the main antibacterial and antibiofilm mechanisms.Discussion: This is the first study that GO and ϵ-PLL coatings were successfully prepared on the surface of CoCrMo alloy by electrodeposition. It provides a promising new approach to the problem of implant infection in orthopedics and stomatology.Keywords: graphene oxide, ϵ-poly-L-lysine, antibacterial, antibiofilm, electroplating

SIRT6, a nuclear protein, has been implicated in a number of essential cellular processes, such as the DNA damage response, metabolic homeostasis, inflammation, tumorigenesis and aging. However, the role of Sirt6 in the regulation of spermatogenesis is yet unknown. In the present study, we successfully generated Sirt6-/- mice on a C57BL6/ICR mixed background and found that some Sirt6-/- mice survived beyond eight weeks. We further revealed that spermatogenesis in Sirt6-/- mice was arrested at the elongated spermatid stage. Sirt6-/- male mice were completely infertile and had an increased number of apoptotic spermatids. To our surprise, deacetylation activities of SIRT6 on H3K9ac, H3K18ac and H3K56c were not required for spermatogenesis. Therefore, our findings establish a novel link between Sirt6 and male fertility, suggesting an essential role of Sirt6 in spermatogenesis.

We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection (micro-TESE-ICSI) treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia (NOA). A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing, China, from January 2014 to December 2017. Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles (Group A). Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles (Group B). We compared patient characteristics, embryonic development, and pregnancy outcomes between Groups A and B. There was no statistical difference in the patient characteristics, and no differences were observed with fertilization or quality embryo rates between Groups A and B. The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B (both P < 0.05). In conclusion, fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.

GGNBP1 is known as gametogenetin protein 1 (GGN1)-interacting protein. It is specifically expressed in the mitochondria of the testis, while its functional role during spermatogenesis is still unknown. Here, we showed that the disruption of Ggnbp1 resulted in abnormal spermiogenesis in around 40% mice, while the others show no defects in the genital system. Moreover, upon treatment with low dose of bisphenol A (BPA), Ggnbp1 knockout mice were more sensitive to environmental pollutant than control mice. The treatment led to decrease in sperm motility and production of abnormal spermatozoa. These results suggest that GGNBP1 mainly ensures proper spermiogenesis in response to various stresses in male mice.

Being infected by SARS-CoV-2 may cause damage to multiple organs in patients, such as the lung, liver and heart. Angiotensin-converting enzyme 2 (ACE2), reported as a SARS-CoV-2 receptor, is also expressed in human male testes. This suggests a potential risk in human male reproductive system. However, the characteristics of ACE2-positive cells and the expression of other SARS-CoV-2 process-related genes are still worthy of further investigation. Here, we performed singlecell RNA seq (scRNA-seq) analysis on 853 male embryo primordial germ cells (PGCs) and 2,854 normal testis cells to assess the effects of the SARS-CoV-2 virus on the male reproductive system from embryonic stage to adulthood. We also collected and constructed the scRNA-seq library on 228 Sertoli cells from three non-obstructive azoospermia (NOA) patients to assess the effects at disease state. We found that ACE2 expressing cells existed in almost all testis cell types and Sertoli cells had highest expression level and positive cells ratio. Moreover, ACE2 was also expressed in human male PGCs. In adulthood, the level of ACE2 expression decreased with the increase of age. We also found that ACE2 positive cells had high expressions of stress response and immune activation-related genes. Interestingly, some potential SARS-CoV-2 process-related genes such as TMPRSS2, BSG, CTSL and CTSB had different expression patterns in the same cell type. Furthermore, ACE2 expression level in NOA donors’ Sertoli cells was significantly decreased. Our work would help to assess the risk of SARS-CoV-2 infection in the male reproductive system. Electronic Supplementary Material Supplementary material is available for this article at 10.1007/s11427-020-1705-0 and is accessible for authorized users.

Objective: Closed malpractice claims against general surgeons were analyzed, to explore causal relationship between surgeons’ behavioral stumble and preventable adverse events.Methods: 127 malpractice claims against general surgeons were reviewed and analyzed. The frequencies of harmful practice patterns of behavior were estimated and it’s relationship with preventability of undesirable outcomes were assessed.Results: 132 legal representatives were involved in the 127 malpractice claims against general surgeons. The top ranks of frequency in flawed behavioral practice patterns were lack of adequate communication with patient and/or family (42.4%), lack of assessment of the primary surgical problem before surgery (34.9%), lack of checking or analyzing investigative report in time (24.2%), lack of evidence of compliance with prevention obligations (18.2%) and lack of following and dealing with postoperatively emerging problems (15.9%). In the involved 132 defendant hospital representatives, it revealed 124 (93.9%) with and 8 (6.1%) without behavioral practice errors. Defendants with behavioral violations were more likely to be considered “surgeons’ errors contributed to the preventable complications” (116 out of 124) than defendants without any behavioral errors (2 out of 8). There was a significant difference between the two groups (P < 0.001).Conclusions: Deficient behavioral practices are ubiquitous in these malpractice claims and play a big role in preventable complications related to general surgery. It is a duty of medical staff leaders or senior surgeons to act as a credible role model ‘in the trenches’ for young generation of surgeons in providing standard of care to their patients mainly through traits of good behavioral practice.

Spermatogenesis is regulated by several Y chromosome-specific genes located in a specific region of the long arm of the Y chromosome, the azoospermia factor region (AZF). AZF microdeletions are the main structural chromosomal abnormalities that cause male infertility. Assisted reproductive technology (ART) has been used to overcome natural fertilization barriers, allowing infertile couples to have children. However, these techniques increase the risk of vertical transmission of genetic defects. Despite widespread awareness of AZF microdeletions, the occurrence of de novo deletions and overexpression, as well as the expansion of AZF microdeletion vertical transmission, remains unknown. This review summarizes the mechanism of AZF microdeletion and the function of the candidate genes in the AZF region and their corresponding clinical phenotypes. Moreover, vertical transmission cases of AZF microdeletions, the impact of vertical inheritance on male fertility, and the prospective direction of research in this field are also outlined.

Purpose This study sought to identify the significant factors related to ongoing pregnancy (OP) and to discover the most reliable model to distinguish OP from non-OP in early gestational age. Methods A total of 1650 cycles were enrolled in this study. Univariate Logistic Regression was used to identify the predictors included in multivariable analysis. The dataset was then randomly split into training set and test set with proportion of 70% and 30%. Forward stepwise multivariable logistic regression with 5-fold cross validation was used to build the final mathematic model. The performance of the model was determined by the arguments of test set. The area under receiver operating characteristic curve (AUC), sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and misclassification rate (MR) were then calculated for model evaluation. Results Seven predictors were related to OP by univariate analysis. The serum hCG level on 14th day post-embryo-transfer (hCG14) and 21th day post-embryo-transfer (hCG21) were linear correlated. Therefore, different multivariate regression models were built using hCG14 or hCG21, respectively. After multivariate regression with 5-fold validation, the final indicators in model-1 were age_group, hCG21 and hCG21/hCG14, while age_group, hCG14, and calculated 48-hour-rising-ratio of hCG were the significant predictors in model-2. Model-2 showed better sensitivity and NPV, lower MR, and similar specificity and PPV. Conclusion This study provided an effective mathematic model for early prediction of OP. The model could be of better clinical significance, especially for clinical counseling to manage patients' stress and anxiety, and for early warning of threatened miscarriage.

Gastric cancer (GC) is one of the major causes of cancer-related mortality. The use of oncolytic virus for cancer gene-virotherapy is a new approach for the treatment of human cancers. In this study, a novel Survivin promoter-driven recombinant oncolytic adenovirus carrying mK5 or MnSOD gene was constructed, which was modified after deletion of the E1B gene. Human plasminogen Kringle 5 mutant (mK5) and manganese superoxide dismutase (MnSOD) are both potential tumor suppressor genes. By constructing Ad-Surp-mK5 and Ad-Surp-MnSOD oncolytic adenoviruses, we hypothesized that the combination of the two viruses would enhance the therapeutic efficacy of GC as compared to the one virus alone. The results of the in vitro experiments revealed that the combination of adenovirus carrying mK5 and MnSOD gene exhibited stronger cytotoxicity to GC cell lines as compared to the virus alone. Additionally, the virus could selectively kill cancer cells and human somatic cells. Cell staining, flow cytometry, and western blot analysis showed that the combination of two adenoviruses containing therapeutic genes could promote the apoptosis of cancer cells. In vivo experiments further verified that Ad-Surp-mK5 in combination with Ad-Surp-MnSOD exhibited a significant inhibitory effect on the growth of GC tumor xenograft as compared to the virus alone, and no significant difference was observed in the bodyweight of treatment and the normal mice. In conclusion, the combination of our two newly constructed recombinant oncolytic adenoviruses containing mK5 or MnSOD therapeutic genes could significantly inhibit gastric cancer growth by inducing apoptosis, suggestive of its potential for GC therapy.

Meiosis is a complex process involving the expression and interaction of numerous genes in a series of highly orchestrated molecular events. Fam9b localized in Xp22.3 has been found to be expressed in testes. However, FAM9B expression, localization, and its role in meiosis have not been previously reported. In this study, FAM9B expression was evaluated in the human testes and ovaries by RT-PCR, qPCR, and western blotting. FAM9B was found in the nuclei of primary spermatocytes in testes and specifically localized in the synaptonemal complex (SC) region of spermatocytes. FAM9B was also evident in the follicle cell nuclei and diffusely dispersed in the granular cell cytoplasm. FAM9B was partly co-localized with SYCP3, which is essential for both formation and maintenance of lateral SC elements. In addition, FAM9B had a similar distribution pattern and co-localization as γH2AX, which is a novel biomarker for DNA double-strand breaks during meiosis. All results indicate that FAM9B is a novel meiosis-associated protein that is co-localized with SYCP3 and γH2AX and may play an important role in SC formation and DNA recombination during meiosis. These findings offer a new perspective for understanding the molecular mechanisms involved in meiosis of human gametogenesis.

Rationale: Idiopathic asthenozoospermia (iAZS) is one of the major causes of male infertility and has no effective therapeutic treatment. Understanding the potential mechanisms that cause it may be helpful in seeking novel targets and treatment strategies for overcoming the problem of low sperm motility in iAZS individuals. Methods: Computer-assisted semen analysis (CASA) was utilized to assess the sperm motility. RT-qPCR, Western blot, immunofluorescence staining, and calcium imaging analysis were performed to examine the expression and function of CatSper channels. Hyperactivation and acrosome reaction were used to evaluate the functional characteristics of epididymal sperm. In vivo fertility assay was applied to determine the fertility of rats. CatSper1 knockdown and overexpression experiments were performed to confirm the roles of CatSper channels in the pathogenesis of iAZS and the therapeutic effects of electroacupuncture (EA) treatment on AZS model rats. Results: Here, we reported a functional down-regulation of CatSper channel from CatSper1 to CatSper 4 in the sperm of both iAZS patients and ornidazole (ORN)-induced AZS model rats, and an impaired sperm function characterized by a reduction of protein tyrosine phosphorylation, hyperactivation, and acrosome reaction in the epididymal sperm of AZS rats. Knockdown of CatSper1 in the testis tissues is sufficient to induce AZS in normal rats, and this action was validated by the reversal effects of CatSper1 overexpression. Transcutaneous electrical acupoint stimulation (TEAS) and electroacupuncture (EA) at 2 Hz frequency improve the sperm motility via enhancing the functional expression of CatSper channels in the sperm. Gene silencing CatSper1 in the sperm abolishes the therapeutic effects of 2 Hz-EA treatment on AZS rats. Conclusions: We conclude that a functional down-regulation of CatSper channel in the sperm may be a contributor or a downstream indicator for a portion of AZS, especially iAZS, while 2 Hz-TEAS or EA treatment has a therapeutic effect on iAZS through inducing the functional up-regulation of CatSper channels in the sperm. This study provides a novel mechanism for the pathogenesis of some AZS especially iAZS, and presents a potential therapeutic target of CatSper for iAZS treatment. Acupuncture treatment like TEAS may be used as a promising complementary and alternative medicine (CAM) therapy for male infertility caused by iAZS in clinical practice.

Aims The intervertebral disc is the largest avascular organ of the body. Vascularization of the disc has been typically regarded as a pathological feature of intervertebral disc degeneration (IDD). However, the underlying mechanism of vascularization in IDD is still unclear. The current study aimed to investigate the role of AF cell derived exosome (AF-exo) in the interaction with human umbilical vein endothelial cells (HUVECs) and its potential role in the regulation of vascularization in IDD. Main methods Human AF tissues were obtained from patients with IDD and idiopathic scoliosis. The AF-exo were isolated and identified by transmission electron microscopy (TEM), nanoparticle trafficking analysis (NTA) and Western blotting. Then, the AF-exo were used for HUVECs cultures. The migration of HUVECs was observed in 2D and 3D cultures. The inflammatory phenotype of HUVECs was examined by Real-time PCR and enzyme-linked immunosorbent assay (ELISA). Additionally, apoptosis of HUVECs were analyzed by flow cytometry. Key findings Here, we for the first time found that AF cells could secrete AF-exo and that the AF-exo could be phagocytosed by HUVECs. Additionally, we found that degenerated AF-exo exerted pro-vascularization effect on HUVECs by promoting cell migration (in 2D and 3D cultures) and inflammatory factor expression including IL-6, TNF-α, MMP-3, MMP-13 and VEGF, whereas the application of non-degenerated AF-exo demonstrated inverse effects. Significance These results showed that AF-exo is an essential regulator mediating intercellular communication between AF cells and HUVECs, suggesting its important role in vascularization in the intervertebral disc.

The aim of our study was to compare the sperm retrieval rates (SRRs) and clinical outcomes of patients with different causes of azoospermia who underwent microdissection testicular sperm extraction-intracytoplasmic sperm injection (micro-TESE-ICSI). We conducted a retrospective study at the Reproductive Medicine Center of Peking University Third Hospital in Beijing, China, from January 2014 to December 2017. This study examined 769 patients with nonobstructive azoospermia who underwent 347 cycles of micro-TESE-ICSI. Patients with azoospermia were classified into Group A (Klinefelter syndrome, n = 284, 125 cycles), Group B (azoospermia Y chromosome factor c [AZFc] microdeletion, n = 91, 64 cycles), Group C (cryptorchidism, n = 52, 39 cycles), Group D (previous mumps and bilateral orchitis, n = 23, 23 cycles), and Group E (idiopathic azoospermia, n = 319, 96 cycles). Clinical characteristics, SRR, embryonic development, and pregnancy outcomes of the patients were compared between all groups. Patients in Group D had the highest and most successful SRR. The average SRR for all patients was 46.0%. The rates of clinical pregnancy, implantation, and live birth in Group D were 78.3%, 65.0%, and 74.0%, respectively, which were higher than those in all other groups (P < 0.05). Group B patients had the lowest clinical pregnancy, implantation, and live birth rates of all groups (P < 0.05). No differences were found in the miscarriage rate or birth defects among the groups (P > 0.05). Patients with orchitis had the highest SRR and best clinical outcomes. Although AZFc microdeletion patients had a higher SRR, their clinical outcomes were worse.

Globozoospermia has been reported to be a rare but severe causation of male infertility, which results from the failure of acrosome biogenesis and sperm head shaping. Variants of dpy-19-like 2 (DPY19L2) are highly related to globozoospermia, but related investigations have been mainly performed in patients from Western countries. Here, we performed a screening of DPY19L2 variants in a cohort of Chinese globozoospermic patients and found that five of nine patients carried DPY19L2 deletions and the other four patients contained novel DPY19L2 point mutations, as revealed by whole-exome sequencing. Patient 3 (P3) contained a heterozygous variant (c.2126+5G>A), P6 contained a homozygous nonsense mutation (c.1720C>T, p.Arg574*), P8 contained compound heterozygous variants (c.1182-1184delATC, p.Leu394_Ser395delinsPhe; c.368A>T, p.His123Arg), and P9 contained a heterozygous variant (c.1182-1184delATCTT, frameshift). We also reported intracytoplasmic sperm injection (ICSI) outcomes in the related patients, finding that ICSI followed by assisted oocyte activation (AOA) with calcium ionophore achieved high rates of live births. In summary, the infertility of these patients results from DPY19L2 dysfunction and can be treated by ICSI together with AOA.

To evaluate and compare left and right testicular tissue histopathology and Johnsen score, and to investigate the necessity for bilateral testicular biopsy. We recruited180 patients with non-obstructiveazoospermia (NOA) on testicular biopsy who had undergonetesticular sperm aspiration (TESA). Pathological sections of testicular tissue were diagnosed by specially-assigned doctors, who evaluated pathological findings, determined the Johnsen score and confirmed for the presence or absence of sperm. Sperm positive rates for left and right testicular histopathology were 55.0% and 51.7% respectively, and the proportion of Johnsen scores≥8 for left and right testes were 53.3% and 50.0%, respectively. Cohen kappa values revealed that the identification of sperm in bilateral testicular samples was not consistent and was related to random effects; Optimized cut-off value for bilateral testicular volume was 11ml (Johnsen score ≥8), and optimized cut-off values of E2 on left and right testes were 144.5pmol/L and 133.5 pmol/L (Johnsen score≤7). However, age, serum prolactin (PRL), follicle stimulating hormone (FSH), luteinizing hormone (LH) and total testosterone (TT) levels were not accurate predictors for the existence of testicular sperm. There was nostatistical significance between left and right testicular histopathology in terms of sperm positive rates or Johnsen score; the Johnsen score were caused entirely by random effects and a score from one side could not represent the other side. Therefore, we recommend that both testes need to undergo surgery when NOA patients undergo testicular biopsy or sperm retrieval.

Acephalic spermatozoa syndrome has been reported for many decades; it is characterized by very few intact spermatozoa and tailless sperm heads in the semen and causes severe male infertility. The only gene in which mutations have been found to be associated with this syndrome encodes Sad1 and UNC84 domain–containing 5 (SUN5), a testis-specific nuclear envelope protein. The functional role of SUN5 has been well-studied in mouse models, but the molecular basis for the pathogenic effects of mutations in the human SUN5 gene remains elusive. Here, we report a new SUN5 mutation (c.475C→T; p.Arg159*), and explore the pathogenic effects of all known SUN5 mutations on acephalic spermatozoa syndrome. Using an artificial splicing system, we found that the intronic mutation affects the splicing of SUN5 mRNA, yielding a premature stop codon that results in a truncated SUN5 protein. We also found that SUN5 interacts with the coupling apparatus protein DnaJ heat shock protein family (Hsp40) member B13 (DNAJB13) during spermatogenesis, and the substitutions in the SUN5 SUN domain impair its interaction with DNAJB13. Furthermore, we observed that many SUN5 mutations affect the secondary structure of the protein and influence its folding and cellular localization. In summary, our findings indicate an interaction of SUN5 with DNAJB13 during spermatogenesis, provide mechanistic insights into the functional role of this interaction in sperm head–tail integration, and elucidate the molecular etiology of acephalic spermatozoa syndrome–associated SUN5 mutations.

Acrolein has been reported to have diverse toxic effects on various organs, including the reproductive system. However, little is known regarding the effects of maternal acrolein exposure on testicular steroidogenesis in male offspring. The present study investigated the effects of acrolein on fetal testosterone production and associated genes. Pregnant Sprague‑Dawley rats were intraperitoneally injected with vehicle (normal saline) or 1, 2 or 5 mg/kg acrolein from gestational day (GD) 14‑20, and fetal testes were examined on GD 21. Fetal body and testicular weights were markedly reduced in pups following exposure to high doses of acrolein (5 mg/kg) in late pregnancy. Notably, in utero exposure of 5 mg/kg acrolein significantly decreased the testicular testosterone level and downregulated the expression levels of steroidogenic acute regulatory protein (StAR) and 3β‑hydroxysteroid dehydrogenase (3β‑HSD), whereas the levels of other steroidogenic enzymes, including scavenger receptor class B, cholesterol side‑chain cleavage enzyme and steroid 17 alpha‑hydroxylase/17,20 lyase, were unaffected. Furthermore, the 3β‑HSD immunoreactive area in the interstitial region of the fetal testes was reduced at a 5 mg/kg dose, whereas the protein expression levels of 4‑hydroxynonenalwere dose‑dependently increased following maternal exposure to acrolein. mRNA expression levels of insulin‑like factor 3, a critical gene involved in testicular descent, were unaltered following maternal acrolein exposure. Taken together, the results of the present study suggested that maternal exposure to high doses of acrolein inhibited fetal testosterone synthesis, and abnormal expression of StAR and 3β‑HSD may be associated with impairment of the steroidogenic capacity.

The Middle Triassic Laojiagou porphyry Mo deposit is located in the Xilamulun metallogenic belt, along the northern margin of the North China Craton (NCC). Molybdenum mineralization is associated with a monzogranite porphyry and occurs primarily in quartz veins and veinlets hosted in wall rocks affected by potassic, silicic, sericitic, and propylitic alteration. The ore-forming process is divided into three stages, characterized by veinlets comprising molybdenite + pyrite + quartz (stage I), pyrite + chalcopyrite + molybdenite + quartz (stage II), and pyrite + quartz (sulfide-poor) (stage III). We identify three fluid inclusion types in quartz grains: carbonic (C-type), liquid-rich two-phase (L-type), and rare vapor-rich two-phase (V-type) inclusions. Microthermometric results indicate that an initial high-temperature (> 350 °C) and low-salinity (

Objective: This study was aimed to evaluate the correlation between clinically significant portal hypertension (CSPH) and postoperative complications and risk predictors of postoperative c...

SDF-1/CXCR4 chemotaxis signals play important roles in regulating the stem cell-based tissue regeneration. The aim of this research is to evaluate whether high expression of CXCR4 enhances the migration of mesenchymal stem cells (MSCs) and increases the efficiency of intervertebral disc (IVD) regeneration. MSCs overexpressing CXCR (CXCR4-MSC) were created by lentiviral-CXCR4-vect transfection, labeled with SPIO, and transplanted into rabbit degenerative IVD induced by annulus puncture. X-ray and T2-weighted MR images of the spine were obtained at 0, 8, and 16 weeks post-transplantation. The transplanted stem cells were traced by both MR imaging and Prussian blue staining. The stem cell-based IVD degeneration was evaluated by quantifying the expression of aggrecan and type II collagen. The in vitro chemotaxis test was performed to study the migration of CXCR4-MSCs to the supplement of SDF-1. The CXCR4-overexpressing MSCs stably elevated the expression of CXCR4 and increased the migration to SDF-1. The SPIO-labeled CXCR4-MSC could be detected within the IVD by MRI till 16 weeks post-transplantation. Prussian blue staining evidenced more SPIO-positive cells within the IVD transplanted with CXCR4-MSCs. Compared to the control group, loss of disc height was slowed while the mRNA expression of aggrecan and type II collagen was increased by MSC transplantation, especially in the IVD supplemented with CXCR4-MSCs. CXCR4 overexpression promoted MSC retention within the IVD and enhanced the stem cell-based IVD regeneration. The SDF-1/CXCR4 chemotaxis signals might help provide a new perspective to understand stem cell migration and infiltration within the degenerated IVD.

Objective: The aim of this study was to investigate the long-term efficacy of laparoscopic radiofrequency ablation (LRFA) in early hepatocellular carcinoma (HCC) compared with other surgical procedures. Methods: A literature search of Cochrane library, PubMed, and Embase through October 2018 was conducted by two investigators (J.-F.G. and F.Y.) independently. The quality of included studies was estimated by the Newcastle-Ottawa Scale. Review Manager 5.3 software was used for meta-analysis, and either fixed- or random-effects model was used according to the heterogeneity of included studies. The chi-square test was used for heterogeneity analysis of included studies, and subgroup analysis was conducted to estimate the heterogeneity between each study and also to estimate the efficacy of different studies. Results: A total of 11 studies involving 1691 patients were included in this analysis. Patients undergoing hepatic resection (HR) had higher 3-, 5-year overall survival rate, 3-year disease-free survival rate, and lower local recurrence rate than those undergoing LRFA. However, patients undergoing LRFA had higher 3-, 5-year overall survival rate than those undergoing other minimally invasive ablation, although there was no statistical difference in local recurrence rate or disease-free survival rate. Conclusion: HR is still an ideal choice for early HCC. If minimally invasive ablation is an alternative treatment, LRFA will be better than other minimally invasive options.

Objectives Traits of metabolic syndrome (MetS) and biomarkers of inflammation and endothelial dysfunction were examined. We investigated the differences of various biomarkers among individuals with or without Mets in a gender-specific manner. The gender-specific associations between E-selectin and MetS were further evaluated. Methods A total of 205 patients were recruited from the outpatient clinics of Tri-Service General Hospital, Taipei, Taiwan. Inclusion criteria were age between 20–75 years and BMI < 35 kg/m2. Demographic, anthropometric and MetS index data were compared between genders. Markers of inflammation and endothelial dysfunction were compared between individuals with or without MetS by gender. Results Age-adjusted E-selectin values showed significant positive correlations with BMI, waist-hip ratio, fasting plasma glucose, systolic and diastolic blood pressure, triglycerides, TNF-α, hsCRP and ICAM-1, and inverse correlation with HDL cholesterol. E-selectin levels were positively correlated with numbers of MetS components in females (P < 0.001) but not in males (P = 0.125). Conclusions Increased E-selectin levels are significantly associated with increased MetS risk in females, but not in males.

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell (ADSC) therapy for erectile dysfunction of cavernous nerve injury (CNI) by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage (2 × 105 cells) of ADSCs for intracavernous injection (ICI) and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs (NanoADSCs) were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.

To investigate the clinical application and outcomes of microdissection testicular sperm extraction (micro-TESE) in patients with nonmosaic Klinefelter syndrome (KS).A total of 143 nonmosaic KS patients underwent micro-TESE in the Center of Reproductive Medicine of Peking University Third Hospital between July 2012 and August 2016. We analyzed their clinical and follow-up data and evaluated the outcomes.Spermatozoa were successfully retrieved from the testicular tissue in 44.76% (64/143) of the patients, 84.4% (54/64) by unilateral and 15.6% (10/64) by bilateral micro-TESE. Seventy-five of the KS patients were followed up in the years of 2014 and 2015. Of the 34 patients with successful sperm retrieval, 73.52% (25/34) achieved clinical pregnancy and 8 boys and 8 girls were already born in 14 of the 25 cases.The micro-TESE is a useful method for sperm retrieval in nonmosaic KS patients, with high rates of sperm retrieval, clinical pregnancy, and birth of biological offspring.目的： 探讨睾丸显微切开取精术在非嵌合型Klinefelter综合征患者中的临床应用及其结局。方法： 总结从2012年7月至2016年8月在我院生殖医学中心行睾丸显微切开取精术的143例非嵌合型Klinefelter综合征患者的临床资料并随访其治疗结局。结果： 143例非嵌合型Klinefelter综合征患者中有64例成功获取精子，精子获得率为44.76%（64/143）；其中顺利获得精子的64例患者中，有84.4%（54/64）患者仅行一侧睾丸显微切开取精即成功获取足够精子，15.6%（10/64）患者需行双侧睾丸显微切开取精获得精子。随访75例患者，成功获取精子的34例患者中，临床妊娠率为73.52%（25/34），已有14例患者妻子成功分娩8个男孩与8个女孩，目前有4例患者妻子正常临床妊娠中。结论： 睾丸显微切开取精术是使非嵌合型Klinefelter综合征患者成功获得自己遗传学子代的有效方法，有较高的精子获得率和临床受孕率。.

To investigate the influence of high fat diet-induced obesity (HFDIO) on the differentially methylated region (DMR) of the imprinted gene and global genome methylation of sperm DNA.We performed bisulfite sequencing on the DMR of the imprinted gene and global genome methylation of sperm DNA in the mouse model of HFDIO.No statistically significant differences were found between the HFDIO model and normal control mice in MEG3-IG (93.73 vs 97.26%, P = 0.252), H19 (98.00 vs 97.83%, P = 0.920), IGF2 (97.34 vs 96.25%, P =0.166), IGF2R (1.43 vs 1.11%, P = 0.695), PEG3 (0.19 vs 0.38%, P = 0.537), MEST (0.23 vs 0.68%, P = 0.315), NNAT (0.31 vs 0.00%, P = 0.134), or SNRPN (1.88 vs 3.13%, P = 0.628). A total of 8 942 DMRs were detected across the sperm genome (P0.05). Gene functional enrichment analysis indicated that the enriched terms with the largest numbers of genes were the metabolic process (n = 1 482), RNA synthesis (n = 779), and transcription (n = 767).The methylation level underwent no significant change in the DMRs of the imprinted genes from the mice with HFDIO, but the CG methylation of the genes involved in the metabolic process, RNA synthesis and transcription were significantly altered.目的： 研究高脂饲料诱导的肥胖是否影响小鼠精子印迹基因差异甲基化区域和精子全基因组DNA甲基化水平。方法： 利用高脂饲料诱导的肥胖小鼠模型，进行精子DNA的全基因组甲基化和印迹基因差异甲基化区测序。结果： 高脂饲料诱导的肥胖组印迹基因差异甲基化区甲基化水平与普通饲料对照组无显著差异：MEG3-IG (93.73% vs 97.26%; P=0.252),H19(98.00% vs 97.83%， P=0.920),IGF2(97.34% vs 96.25%，P=0.166),IGF2R(1.43% vs 1.11%，P=0.695), PEG3(0.19% vs 0.38%，P=0.537),MEST(0.23% vs 0.68%，P=0.315), NNAT(0.31% vs 0.00%，P=0.134)和SNRPN(1.88% vs 3.13%，P=0.628)。精子全基因组范围内总共发现8 942个甲基化有显著差异的区域(P0.05)。对差异甲基化区域进行基因功能富集，富集基因数目最多的3项分别为代谢(n=1 482)，RNA合成(n=779)和转录(n=767)。结论： 高脂饲料诱导的肥胖小鼠精子印迹基因差异甲基化区甲基化水平没有发生显著改变，参与代谢、RNA合成以及转录过程基因CG序列甲基化显著改变。.

To study the effect of transcutaneous electrical acupoint stimulation (TEAS) in the treatment of asthenozoospermia.We randomly divided 72 asthenozoospermia patients into a 2 Hz TEAS (n = 29), a 100 Hz TEAS (n = 20), and a blank control group (n = 23), those in the former two groups treated by 30 minutes of TEAS at 2 Hz and 100 Hz respectively, applied to the acupoints of bilateral Shenshu, left Zusanli, and Guanyuan, once a day for 60 days, while those in the blank control group left untreated. Using computer-assisted sperm analysis (CASA), we examined sperm concentration and motility as well as the percentages of grade a and grade a+b sperm in different groups of the patients.Compared with the baseline, 2 Hz TEAS significantly increased sperm motility (［12.76 ± 1.39］ vs ［18.89 ± 2.46］%, P0.05) and the percentage of grade a+b sperm ( ［10.68 ± 1.22］ vs ［16.32 ± 2.10］%, P0.05) in the asthenozoospermic patients, while 100 Hz TEAS improved not only sperm motility (［12.32 ± 2.21］ vs ［23.81 ± 3.42］%, P0.01) and the percentage of grade a+b sperm (［10.45 ± 1.98］ vs ［20.25 ± 2.82 ］%, P0.01), but also the percentage of grade a sperm (［6.44 ± 1.16］ vs ［13.31 ± 2.30］%, P0.05). Moreover, in comparison with the blank control group, 2 Hz TEAS also remarkably increased sperm motility (［9.57 ± 1.60］ vs ［18.89 ± 2.46］%, P0.05) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［16.32 ± 2.10］%, P0.05) in the asthenozoosperma patients, while 100 Hz TEAS improved not only sperm motility (［9.57 ± 1.60］ vs ［23.81 ± 3.42］%, P0.01) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［20.25 ± 2.82］%, P0.01) but also the percentage of grade a sperm (［4.87 ± 1.01］ vs ［13.31 ± 2.30］%, P0.01). Meanwhile, the rate of clinical effectiveness was significantly higher in the 100 Hz TEASthan in the blank control group either in intention-to-treat (ITT) analysis (100% vs 18.18%) orper-protocol (PP) analysis (90% vs 0%), and so was it than in the 2 Hz TEAS group based on the data of ITT (100% vs 33.33%).Both 2 Hz and 100 Hz TEAS are effective for the treatment of asthenozoospermia by improving sperm motility and vitality.目的： 研究经皮穴位电刺激(TEAS)对弱精子症患者的治疗效果。方法： 将72例弱精子症患者随机分为：2 Hz TEAS治疗组（29例）、100 Hz TEAS治疗组 （20例）及空白对照组（23例）。TEAS治疗组选取双侧肾俞穴、左侧足三里穴及关元穴4个穴位；TEAS参数：施以连续波，频率2 Hz或100 Hz，每日1次，每次30 min 。空白对照组不采取任何治疗手段，时间均为60 d。通过计算机辅助精子分析(CASA)观察2 Hz及100 Hz TEAS治疗对弱精子症患者的精子活动率、浓度、a级精子百分 率及a+b级精子百分率等指标的影响。 同时采用意向治疗分析(ITT)和符合方案数据分析(PP)对临床疗效进行评判。结果： 与治疗前比较，2 Hz TEAS治疗可显著提高 弱精子症患者的精子活动率［(12.76 ± 1.39)% vs (18.89 ± 2.46)%, P0.05］及a+b级精子百分率［(10.68 ± 1.22)% vs （16.32 ± 2.10）%, P0.05］；100 Hz TEAS治疗不仅能够提高弱精子症患者的精子活动率［(12.32 ± 2.21)% vs (23.81± 3.42)%, P0.01］及a+b级精 子百分率［(10.45 ± 1.98)% vs (20.25 ± 2.82)%, P0.01］，而且还能够提高a级精子百分率［(6.44 ± 1.16)% vs (13.31 ± 2.30)%, P0.05］。此外，2 Hz TEAS治疗组的精子活动率［(9.57 ± 1.60)% vs (18.89 ± 2.46)%, P0.05］及a+b级精子百分率［(7.81 ± 1.31)% vs (16.32 ± 2.10)%, P0.05］显著高于空白对照组治疗后；100 Hz TEAS治疗组的精子活动率［(9.57 ± 1.60)% vs (23.81 ± 3.42)%, P0.01］、a+b级精子百分率［(7.81 ± 1.31)% vs (20.25 ± 2.82)%, P0.01］及a级精子百分率［(4.87 ± 1.01)% vs (13.31 ± 2.30)%, P0.01］亦均显著高于空白对照组治疗后。 同时100 Hz TEAS的临床疗效无论是在ITT数据集( 100% vs 18.18%)，还是PP数据集中( 90% vs 0%) 均显著高于空白对照组；而在ITT数据集中，100 Hz TEAS的临床疗效(100% vs 33.33%)亦显著高于2 Hz TEAS。 结论： 2 Hz、100 Hz经皮穴位电刺激均可以提高弱精 子症患者的精子活动率和活力，对弱精子症有一定的治疗作用。.

Krukenbergtumor is a metastatic ovarian tumor with its primary site being the gastrointestinal tract. The pathogenesis of Krukenberg tumor formation is still in its hypothetical stage though the current understanding suggests lymphatic, hematogenous and transcoelomic route as the 3 major route of metastasis. There is a lack of description in the literature related to the pathway of metastasis. Here, we intend to search the available literature and provide a thorough review, which may be helpful to the readers to understand the issue of mechanism of Krukenberg tumor metastasis more clearly.