Case report: remedial microdissection testicular sperm extraction after onco-microdissection testicular sperm extraction failure.

BACKGROUND: Testicular cancer (TC) mostly occurs in men aged 14 to 44. Studies have shown that TC seriously damages male fertility, and 6% to 24% of patients with TC were even found to suffer from azoospermia when they are diagnosed. At present, some studies have pointed out that onco-microdissection testicular sperm extraction (mTESE) can extract sperm from tumor testicles. However, there are almost no reports on remedial measures after onco-mTESE failure. Given the valuable opportunity for fertility preservation in patients with TC and azoospermia, it is necessary to provide effective remedial methods for patients with failed onco-mTESE. METHODS: Two young men, who were diagnosed with TC and also found to have azoospermia, tried onco-mTESE while undergoing radical orchiectomy for fertility preservation. However, sperm extraction failed in both patients. Subsequently, the isolated testicular tissue of the patient in case 1 suffered from TC again, and the patient in case 2 was scheduled to receive multiple cycles of gonadotoxic chemotherapy. Because both had a plan to have a birth in the future, we performed remedial mTESE. RESULTS: Sperm was successfully extracted from both patients. The patient recovered well, without complications. The patient couple in case 1 underwent 1 intracytoplasmic sperm injection (ICSI) cycle but did not achieve clinical pregnancy. CONCLUSIONS: There is still an opportunity to extract sperm successfully using onco-mTESE, despite the difficulty of fertility preservation in TC patients with azoospermia. If sperm extraction from the tumor testis fails, implementing remedial mTESE as early as possible would likely preserve the last chance of fertility for these patients.

Normal sperm head morphometric reference values in fertile Asian males.

ABSTRACT: Sperm head morphology is crucial for male factor infertility diagnosis and assessment of male reproductive potential. Several criteria are available to analyze sperm head morphology, but they are limited by poor methodology comparability and population applicability. This study aimed to explore comprehensive and new normal morphometric reference values for spermatozoa heads in fertile Asian males. An automated sperm morphology analysis system captured 23 152 stained spermatozoa from confirmed fertile males. Of these samples, 1856 sperm head images were annotated by three experienced laboratory technicians as "normal". We employed 14 novel morphometric features to describe sperm head size (head length, head width, length/width ratio, and girth), shape (ellipse intersection over union, girth intersection over union, short-axis symmetry, and long-axis symmetry), area (head, acrosome, postacrosomal areas, and acrosome area ratio), and degrees of acrosome and nuclear uniformity. This straight-forward method for the morphometric analysis of sperm by accurate visual measurements is clinically applicable. The measured parameters present valuable information to establish morphometric reference intervals for normal sperm heads in fertile Asian males. The presented detailed measurement data will be valuable for interlaboratory comparisons and technician training. In vitro fertilization and andrology laboratory technicians can use these parameters to perform objective morphology evaluation when assessing male fertilization potential.

External quality assessment scheme for sperm DNA fragmentation: a pilot study in China.

BACKGROUND: The aim of this article is to establish an external quality assessment (EQA) scheme for sperm Deoxyribonucleic acid (DNA) fragmentation (SDF) detection, and to assess the feasibility of the scheme. In addition, this article provides some case analysis of abnormal results in order to really help improve the performance of the laboratory. RESULTS: In 2021 and 2022, 10 and 28 laboratories in China volunteered to participate in the EQA program respectively. Two samples were selected for EQA each year, a large spread of results was obtained for the four samples, and the highest values were 13.7, 4.2, 8.0 and 4.0 times the lowest respectively. The coefficients of variation (CVs) were very high for the four samples, at 46.6%, 30.1%, 26.7% and 30.3%, respectively. The CVs of the samples with high SDF values were lower than those of the samples with low SDF values. There was no significant difference between the results of sperm chromatin structure assay (SCSA) and sperm chromatin dispersion (SCD). For the 10 laboratories that participated in EQA in 2021 and 2022, the CVs of low SDF value samples and high SDF value samples decreased from 46.6% and 30.1% in 2021 to 32.5% and 22.7% in 2022, respectively. CONCLUSION: This is the first study to evaluate the EQA program on SDF, which involved a number of laboratories and was demonstrated to be feasible. It is recommended that all laboratories participate in the EQA of SDF to ensure the accuracy of the results.

RéSUMé: CONTEXTES: L'objectif de cet article est d'établir un système externe d'évaluation de la qualité (EEQ) pour la détection de la fragmentation de l'ADN des spermatozoïdes (SDF) et d'évaluer la faisabilité de ce système. En outre, cet article fournit une analyse de cas de résultats anormaux afin d'aider réellement à améliorer les performances du laboratoire. RéSULTATS: En 2021 et 2022, respectivement 10 et 28 laboratoires en Chine se sont portés volontaires pour participer au programme EEQ. Deux échantillons ont été sélectionnés chaque année pour l'EEQ ; un large éventail de résultats a été obtenu pour les quatre échantillons, et les valeurs les plus élevées étaient respectivement de 13,7, 4,2, 8,0 et 4,0 fois les plus faibles. Les coefficients de variation (CV) étaient très élevés pour les quatre échantillons, soit respectivement 46,6 %, 30,1 %, 26,7 % et 30,3 %. Les CV des échantillons avec des valeurs de SDF élevées étaient inférieurs à ceux des échantillons avec de faibles valeurs de SDF. Il n'y avait pas de différence significative entre les résultats du test de structure de la chromatine des spermatozoïdes (SCSA) et ceux de la dispersion de la chromatine des spermatozoïdes (SCD). Pour les 10 laboratoires qui ont participé à l'EEQ en 2021 et 2022, les CV des échantillons à faible valeur de SDF et ceux des échantillons à valeur élevée de SDF ont diminué, passant respectivement de 46,6 % et 30,1 % en 2021 à 32,5 % et 22,7 % en 2022. CONCLUSIONS: Il s'agit de la première étude à évaluer le programme externe d'évaluation de la qualité (EEQ) de l'analyse de la SDF, qui a impliqué un certain nombre de laboratoires, et qui s'est avéré réalisable. Il est recommandé que tous les laboratoires participent à l'EEQ de la SDF afin d'en assurer l'exactitude des résultats.

Overexpressed SNRPB/D1/D3/E/F/G correlate with poor survival and immune infiltration in hepatocellular carcinoma.

BACKGROUND: Prior reports have indicated that the abnormal expression of small nuclear ribonucleoproteins (snRNPs) genes is related to malignant tumors. However, in hepatocellular carcinoma (HCC), the precise role of snRNPs is not well understood. Therefore, the purpose of this study was to evaluate the prognostic roles of SNRPB/D1/D2/D3/E/F/G and their correlation to immune infiltration in HCC. METHODS: The study was carried out via the following databases, software, and experimental validation: ONCOMINE, GEPIA2, UALCAN, The Cancer Genome Atlas, Gene Expression Omnibus, ArrayExpress, Kaplan-Meier plotter, cBioPortal, STRING, DAVID 6.8, TIMER, Cytoscape software, and immunohistochemistry experiments. RESULTS: Overexpressed SNRPB/D1/D2/D3/E/F/G proteins were found in HCC tissues. The transcription levels of 7 snRNPs genes were related to the TP53 mutation and tumor grades. SNRPB/D1/D2/D3/F/G expression was significantly correlated with cancer staging, whereas SNRPE was not. Moreover, Kaplan-Meier survival analysis showed that upregulation of SNRPB/D1/D2/E/G was relevant to worse OS in HCC patients, especially in patients with alcohol consumption and those without viral hepatitis. Multivariate Cox regression analysis indicated that expression of SNRPB/D1/D3/E/F/G were independent prognostic factors for unfavorable OS in HCC. In addition, a high mutation rate of snRNPs genes (44%) was also found in HCC. The mRNA expression levels of snRNPs were meaningfully and positively related to six types of infiltrating immune cells (B cells, CD4+ T cells, CD8+ T cells, neutrophil, macrophage, and dendritic cells). Also, SNRPB/D1/G genes were significantly associated with molecular markers of various immune cells in HCC. CONCLUSIONS: SNRPB/D1/D3/E/F/G are potential prognostic biomarkers for a short OS in HCC, and SNRPB/D1/G were novel immune therapy targets in HCC patients.

Combined effects of green manure returning and addition of sewage sludge compost on plant growth and microorganism communities in gold tailings.

Remediation of gold tailings is often difficult due to their extremely barren nature and highly heavy metal concentrations. Returning green manure and applying sewage sludge compost have the beneficial effects of providing nutrients and improving the soil environment. The effects of green manure plants, alfalfa (Medicago sativa L.), ryegrass (Lolium perenne Linn.), and tall fescue (Festuca arundinacea), returning in situ on nutrients, bioavailability of trace metals, and community structure of microorganism in gold tailings amended with 0%, 5%, and 10% (weight/weight) sewage sludge compost on the top 4 cm of tailings (SSC-5, SSC-10) were investigated in a pot experiment. The results showed that the plant biomass and microbial biomass carbon in tailings significantly increased in the treatments with sewage sludge compost. The available N and available P and the availability of Zn decreased markedly with the returning of alfalfa and ryegrass. Moreover, through high-throughput sequencing, it was found that the returning of alfalfa had positive effects on the bacterial community richness but a negative impact on the fungal community richness. The microbial community diversity was reduced in the treatment without sewage sludge compost amendment and with alfalfa returning. However, the microbial community diversity was enriched in the treatment of alfalfa returning with sewage sludge compost. In each plant species, 9 dominant bacterial phyla and 10 dominant fungi phyla could be detected. Returning alfalfa green manure and applying sewage sludge compost led to a relative increase in the abundance of Proteobacteria and Ascomycota. These results demonstrated that returning alfalfa and applying sewage sludge compost could be effective in the ecological restoration of gold tailings.

[Advantages of cryopreservation technology for rare human sperm].

OBJECTIVE: To search for an effective method for cryopreservation of rare human sperm (RHS) by comparing the effect of RHS cryopreservation technology with that of conventional cryopreservation technology on post-thaw sperm from patients with severe oligozoospermia. METHODS: Semen samples obtained from 82 patients with severe oligozoospermia were preserved by RHS cryopreservation technology, and another 24 samples cryopreserved by conventional technology, the former divided into groups A (sperm concentration < 1×106/ml, n = 54) and B (1×106/ml ≤ sperm concentration < 5×106/ml, n = 28), and the latter included in group C (sperm concentration < 15×106 /ml, n = 24). The survival rate of post-thaw sperm and recovery rate of progressively motile sperm (PMS) were compared among the three groups. RESULTS: The survival rate of post-thaw sperm was significantly higher in groups A and B than in C (ï¼»62.8 ± 18.7ï¼½% and ï¼»61.9 ± 17.2ï¼½% vs ï¼»50.7 ± 13.5ï¼½%, P < 0.05), and so was the recovery rate of PMS (ï¼»68.7 ± 18.4ï¼½% and ï¼»70.7 ± 15.5ï¼½% vs ï¼»29.2 ± 12.4ï¼½% , P < 0.05), but there were no statistically significant differences between groups A and B in either of the two parameters (P > 0.05). CONCLUSIONS: The cryopreservation technology for rare human sperm may yield relatively stable post-thaw results and deserves a wide clinical application in preserving male fertility.

[Levels of T-Lymphocyte Subsets, IL-17, IL-35 and IFN-γ in Peripheral Blood and Their Clinical Significance in Patients with Multiple Myeloma].

OBJECTIVE: To explore the levels of T-lymphocyte subsets and IL-17, IL-35, IFN-γ in peripheral blood of patients with multiple myeloma(MM) and their clinical significance. METHODS: A total of 86 MM patients in our hospital from January 2014 to January 2017 were enrolled in MM group and 30 healthy persons were enrolled in control group, the CD4+/CD8+ T cells ratio, CD4+CD25high/+ CD127low/- Treg level in peripheral blood were detected by flow cytometer. The levels of IL-17, IL-35 and IFN-γ in peripheral serum were detected by ELISA, and the differences of detected indicators between different groups were compared. RESULTS: Compared with control group, the proportion of CD4+ T cells and CD4+/CD8+ T cells ratio decreased, the proportion of CD8+ T cells and Treg increased in MM group. The differences of T lymphocyte subsets level between group III stage of MM and control group were statistically significant (P<0.05). With enhancing of clinical stages, Treg level showed a increasing trend, especially in III stage (P<0.05), the serum level of IL-17 as followed in turn: III stage>II stage>I stage>control, the serum level of IL-35 and IFN-γ as followed in turn: control>I stage>II stage>III stage (P<0.05). In terms of disease status, the propurtion of Treg cells as fllowed in turn: disease progression stage>stable stage>control (P<0.05), the serum level of IL-17 as followed in turn also: disease progression stage>stable stage (P<0.05), while the serum level of IL-35 and IFN-γ as followed in turn: control>disease table stage>progression stage (P<0.05). CONCLUSION: The abnomal level of T-lymphocyte subsets, Treg, IL-17, IL-35 and IFN-γ are related with progression and prognosis of MM patients.

The influences of arbuscular mycorrhizal fungus on phytostabilization of lead/zinc tailings using four plant species.

A greenhouse experiment was conducted to investigate the effects of the arbuscular mycorrhizal fungus Funneliformis mosseae on three parameters: Pb, Zn, Cu and Cd accumulation, translocation and plant growth in perennial ryegrass (Lolium perenne), tall fescue (Festuca arundinacea), showy stonecrop (Hylotelephium spectabile) and Purple Heart (Tradescantia pallida). The purpose of this work is to enhance site-specific phytostabilization of lead/zinc mine tailings using native plant species. The results showed that mycorrhizal fungi inoculation significantly increased plant biomass of F. arundinacea, H. spectabile and T. pallida. The Pb, Zn, Cu and Cd concentrations in roots were higher than those in shoots both with and without mycorrhizae, with the exception of the Zn concentration in H. spectabile. Mycorrhizae generally increased metal concentrations in roots and decreased metal concentrations in shoots of L. perenne and F. arundinacea. In addition, it was found that the majority of the bioconcentration and translocation factors were lower than 1 and mycorrhizal fungi inoculation further reduced these values. These results suggest that appropriate plant species inoculated with mycorrhiza might be a potential approach to revegetating mine tailing sites and that H. spectabile is an appropriate plant for phytostabilization of Pb/Zn tailings in northern China due to its higher biomass production and lower metal accumulation in shoots.

[Short-term effect of black film covering on Oncomelania hupensis snail control in marshland and lake regions].

OBJECTIVE: To evaluate the short-term effect of the black film covering combined with niclosamide on Oncomelania hupensis snail control in marshland and lake regions. METHODS: A ditch with snails in Anxiang County was selected as the experimental sites. The ditch was divided into 3 groups: a film group, a niclosamide group, and a niclosamide and film group. The snails were surveyed before the test and 1, 3, 5, 7 days after the experiment. RESULTS: After the film covering, the highest temperature of the surface of soil was 63.7 degrees C on the first day, and the temperature higher than 40 degrees C lasted 4.34 hours. On the second day, the highest temperature was 52.3 degrees C, and the temperature higher than 40 degrees C lasted 4.96 hours. On the fourth day and fifth day, the temperature was lower than 30 degrees C, while the temperature was lower than 34 degrees C on the sixth day. On the seventh day of the experiment, the snail death rates of the film group, the niclosamide group, and the niclosamide and film group on the soil surface were 16.36%, 58.40% and 53.57%, respectively. On the seventh day, the snail death rates of the film group, and the niclosamide and film group under the soil were 20.00% and 40.00%, respectively, while no snails were found under the soil in the niclosamide group 3 days after the experiment. In mesh bags, the snail death rates of the film group, the niclosamide group, and the niclosamide and film group were 84.00%, 95.33% and 95.33%, respectively. CONCLUSION: The short-term effect of the black film covering on snail control is not obvious, and the black film covering does no promote the molluscicidal effect of niclosamide.

Endovascular stent graft placement in the treatment of a ruptured tuberculous pseudoaneurysm of the descending thoracic aorta secondary to Pott's disease of the spine.

Ruptured thoracic aortic tuberculous pseudoaneurysms as a complication of mycobacterium tuberculosis infection of the spine are rare. Conventional treatment of a ruptured tuberculous pseudoaneurysm involves surgery with graft interposition or patch repair. We report successful repair of a ruptured tuberculous pseudoaneurysm of the descending thoracic aorta by endovascular stent graft placement and provide a literature review of such entities.

A 23-year-old male with Uhl's anomaly.

We report a case of a 23-year-old male with Uhl's anomaly who presented with nonparoxysmal atrioventricular junctional tachycardia with Mobitz I. The patient underwent successful total cavopulmonary conversion.

Myocardial lipomatous infiltration of the left ventricular wall.

A myocardial mass was found on the left ventricle wall of a four-year-old Chinese girl diagnosed with pulmonary valve stenosis and patent foramen ovale. Surgical valvotomy and partial excision of the myocardial mass were performed, and a biopsy of the myocardial mass revealed myocardial lipomatous infiltration.

[Xianling Gubao Capsule improves semen quality].

OBJECTIVE: To observe the effect of the compound Chinese traditional drug Xianling Gubao Capsule on the semen quality of infertile males. METHODS: We treated 66 infertile men with Xianling Gubao Capsule for 24 months, and analyzed the semen quality and sperm morphology before and after the treatment. RESULTS: Two months after the medication, sperm concentration was increased by a small margin, but no statistically significant changes were observed in sperm vitality and motility (P > 0.05), the rate of morphologically normal sperm was significantly raised from 25.8% before treatment to 57.6% (P < 0.05) in those with the normal rate > or = 15%, but decreased from 53.0% to 25.8% (P < 0.05) in those with the normal rate < 9%. Among the 7 cases of oligospermia, the rate of morphologically normal sperm was elevated to an average of 10.9% after the 4-month medication, significantly different from the baseline rate of 5.8% (P < 0.05). Five spontaneous pregnancies and 1 successful IVF-ICSI were achieved during the treatment. CONCLUSION: Xianling Gubao Capsule can improve semen quality and significantly increase the percentage of morphologically normal sperm.

[Antimicrobial resistance and molecular epidemiology of coagulase-negative Staphylococci in Chengdu].

OBJECTIVE: To investigate the antimicrobial resistance and molecular prevalence of coagulase-negative Staphylococcus (CONS) in Chengdu. METHODS: Disk diffusion method was used to test antimicrobial resistance of 2038 CoNS isolated from 15 hospitals and 6 CDCs (center for disease control and prevention) in Chengdu, the results were interpreted according to the guideline of National Committee for Clinical Laboratory Standards. CoNS were typed with plasmid and PFGE (pulsed field gel electrophoresis) profiles. RESULTS: (1) S. epidermidis, S. haemolyticus, and S. saprophyticus accounted for 82.2% of the CoNS isolated from Chengdu, with different constituent ratio of CoNS among patients, medical staff, sick children and normal people. (2) CoNS resisted to one or more antimicrobial agents with resistance rate of 80.4% . CoNS resisted highly to SMZ, penicillin-G, ampicillin, erythromycin, TMP-SMZ and tetracycline, but were susceptible to vancomycin, norfloxacin and amikacin. 25 antimicrobial resistance profiles were acquired, and Amp + Ery + P-G + SMZ, Amp + Gen + Str + Tet, Amp + P-G + Tet, Chl + Ery + P-G + SMZ + TS, Ern + Nov + P-G + SMZ + TS, as well as P-G + SMZ + TS were main profiles. The main antimicrobial resistance profiles of CoNS isolated from patients, medical staff, sick children and normal people had some differences, but the antimicrobial resistance of main biochemical subtypes was similar a lot. (3) Plasmid prevalence of CoNS was 72.9%, with 12 plasmid profiles ( I -H ) of all CoNS. The main profiles were I , U , and MI type, which accounted for 80.1% of 1485 CoNS with plasmid. (4) 29 PFGE genotypes and 112 subtypes were found in 2038 strains. Genotypes A, B, C, D and E were the predominant types in CoNS from patients, medical staff, sick children and normal people and contributed 89.1% to 2038 CONS. Genotypes A was the major type and had similar constituent ratio in CoNS from 4 sources, and no enough similarity of constituent ratio in other dominant genotypes. CONCLUSION: Antimicrobial resistance rate of CoNS was very high in Chengdu and multi-resistance existed in various populations generally. High homology was found in CoNS from different sources, especially from medical staff as well as sick children and infant, and a latent inter-transmission may occur in these populations. So it was no time to delay the monitoring and investigation of antimicrobial resistance and molecular epidemiology for CoNS in Chengdu.

[Study on the resistance of Escherichia coli O157:H7 to disinfectants in relationship with its plasmid pO157 and chromosome DNA].

OBJECTIVE: To study the change of Escherichia coli O157:H7 resistance to disinfectants in continuous disinfection and the relationship between the resistance and pO157, chromosome DNA. METHODS: Using 4 disinfectants in a separate manner to disinfect 5 Escherichia coli O157:H7 50 generations continuously, we made a before-after comparison of their resistibility and analyzed the change of the structure of pO157 and chromosome DNA. RESULTS: After the 50-generation-continuous disinfection, the bacteria resistance to sodium dichloroisocyanurate, iodophor and quaternary ammonium increased, but the resistance to chlorhexidiniacetas did not any change; the maps Cal I and Rsr II cutting pO157 revealed some changed after disinfection by sodium dichloroisocyanurate and quaternary ammonium, but the maps showed no change after disinfection by iodophor and chlorhexidiniacetas; the chromosome DNA PFGE maps change considerably after 50 generations of disinfection by sodium dichloroisocyanurate, iodophor and quaternary ammonium, but the chromosome DNA PFGE maps were similar after 50 generation of disinfection by chlorhexidiniacetas. CONCLUSION: The Escherichia coli O157:H7 resistance to disinfectants will increase after the 50-generation-continuous disinfection by sodium dichloroisocyanurate, iodophor and quaternary ammonium. There may be genes both in chromosome DNA and in pO157 which resist sodium dichloroisocyanurate and quaternary ammonium, the reason for increase of resistance may be related with the change of chromosome DNA and pO157. There may be genes in chromosome DNA which resist iodophor, the reason for increase of resistance to iodophor may be related with the change of chromosome DNA, but not related with pO157.

[Influence of cryoprotectant with glycerol and freezing-thawing procedure on the motility of human sperm].

OBJECTIVE: To investigate the effects of cryoprotectant with glycerol and the freezing-thawing procedure on the motility of METHODS: The motion characteristics of human sperm from 18 selected specimens were assessed by computer-assisted human sperm. sperm analysis before and after adding hyper-osmolarity cryoprotectant with glycerol and the freeze-thaw procedure, and the data were evaluated in pairs. RESULTS: The adding of cryoprotectant caused an increase in the proportion of rapid linear motile sperms (P < 0.05) and sperm velocities, including VCL, VSL and VAP (P < 0.005). But no changes were observed in the proportion of progressively motile sperms and sperm motility. Compared with the data from pre-frozen samples, velocities of post-thawed sperms and the percentage of motile sperms in each grade significantly declined (P < or = 0.01), so did ALH, while WOB, LIN and STR remained unchanged (P > 0.05). Significant differences in ALH, WOB, LIN and STR were observed only in comparison between the post-thawed and pre-treated samples. CONCLUSION: The number of Grade a sperms and sperm velocity increased after adding hyper-osmolarity cryoprotectant with glycerol. The sperm motile potential was impaired and even entirely destroyed in some cases by cryodamage. The influences on the sperm motion were differently induced in freezing-thawing procedures.

[Semen quality and sperm morphologic study of infertile men with varicocele].

OBJECTIVE: To examine the semen quality and the sperm morphology in infertile men with varicocele. METHODS: Semen from 98 infertile men with varicocele were studied and those of 130 normal semen donors were taken as the control. Semen analysis was performed based on the methods described in the WHO manual and sperm morphology was evaluated by WHO criteria. RESULTS: A significantly reduced percentage of normal morphologic sperm and of forward progression were found in patients with varicocele comparing with those of the control (P <0.001). The head defects were observed as the predominant type of sperm malformation. CONCLUSION: The varicocele increases malformed sperm in ejaculates, which may result from impaired male fertility by varicocele. Sperm morphologic assessment with WHO criteria provides a sensitive and practical measurement of sperm damage in infertile men with varicocele.