Biallelic mutations in spermatogenesis and centriole-associated 1 like (SPATC1L) cause acephalic spermatozoa syndrome and male infertility / 亚洲男科学杂志(英文版)

Acephalic spermatozoa syndrome is a rare type of teratozoospermia that severely impairs the reproductive ability of male patients, and genetic defects have been recognized as the main cause of acephalic spermatozoa syndrome. Spermatogenesis and centriole-associated 1 like (SPATC1L) is indispensable for maintaining the integrity of sperm head-to-tail connections in mice, but its roles in human sperm and early embryonic development remain largely unknown. Herein, we conducted whole-exome sequencing (WES) of 22 infertile men with acephalic spermatozoa syndrome. An in silico analysis of the candidate variants was conducted, and WES data analysis was performed using another cohort consisting of 34 patients with acephalic spermatozoa syndrome and 25 control subjects with proven fertility. We identified biallelic mutations in SPATC1L (c.910C>T:p.Arg304Cys and c.994G>T:p.Glu332X) from a patient whose sperm displayed complete acephalia. Both SPATC1L variants are rare and deleterious. SPATC1L is mainly expressed at the head-tail junction of elongating spermatids. Plasmids containing pathogenic variants decreased the level of SPATC1L in vitro. Moreover, none of the patient's four attempts at intracytoplasmic sperm injection (ICSI) resulted in a transplantable embryo, which suggests that SPATC1L defects might affect early embryonic development. In conclusion, this study provides the first identification of SPATC1L as a novel gene for human acephalic spermatozoa syndrome. Furthermore, WES might be applied for patients with acephalic spermatozoa syndrome who exhibit reiterative ICSI failures.

The physiological and psychological mechanisms of infra-slow oscillation / 生理学报

Infra-slow oscillation (ISO) is a kind of brain rhythm between 0.01 and 0.5 Hz. ISO is widely distributed in multiple brain regions. As an important psychophysiological activity, the ISO interacts with high-frequency neural rhythm via cross-frequency coupling, but has different activity patterns from high-frequency neural activity. Physiologically, the ISO may be generated by the dynamic activity of thalamus, glia, and ions. Psychologically, the frequency, amplitude, and phase of ISO could all regulate cognitive activities, but in different ways. Investigations on the ISO expands the neural rhythm research to lower frequency range, further promoting the construction of rhythmic theory of brain function.

The Establishment of SCID Mice Model on Endometriosis / 昆明医科大学学报

Objective To set up a favorable animal model for the drug treatment research of endometriosis by establishing the animal model of endometriosis in SCID and nude mice so as to compare the influences on implantation of human endometrial tissue derived from the eutopic and ectopic sources. Methods Eutopic and ectopic endometrium were transplanted to the lower abdominal parts subcutaneously of 30 sexually matured BALB/c-nu/nu nude mice and SCID mice respectively. The ectopic lesion sizes were under the regular observation before they were removed 6 weeks after the operation for pathological examinations. Results Nude mice and SCID mice were able to be used to establish a successful animal models of endometriosis. The success rate of SCID mice was higher than that of nude mice. The success rate of the eutopic endometrium group was significantly higher than that of the ectopic endometrium group. Nude and SCID mice endometriosis implantation models were successfully established. The modeling success rate of SCID mice is higher than that of the nude mice.The success rate of transplantation was higher in the ectopic endometrium than in the eutopic endometrium.Conclusion The SCID mice endometriosis endometriosis model provides a favorable animal model of endometriosis.

Total laparoscopic radical trachelectomy with ascending branches of uterine arteries preservation / 中华医学杂志(英文版)

Radical trachelectomy (RT) is a fertility-sparing treatment for young women with early-stage cervical cancer. We report here a case of a 30-year-old nulliparous woman who presented with stage IA2 cervical squamous cancer. She was treated with total laparoscopic radical trachelectomy (TLRT) and laparoscopic pelvic lymphadenectomy (LPL). During this procedure, the ascending branches of uterine arteries were preserved. No metastasis was identified after fourteen months of follow-up. The menstrual pattern normalized and the patient has been attempting to conceive for two months. TLRT might be a safe fertility-preserving procedure for early-stage cervical cancer, due to its minimally invasive nature and shorter recovery time. However, more data are required on recurrence rate, fertility rate and pregnancy outcome in order to fully evaluate the therapeutic efficacy of TLRT.

Making of the animal model with sterilized testes / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the methods of making an animal model with sterilized testes.</p><p><b>METHODS</b>(1) X-ray local irradiation. Seventy 8-10-week-old male mice were equally divided into 6 experiment groups and a control group. The testes of the mice in the 6 experiment groups were irradiated sequentially by 1000, 1200, 1400, 1600, 1800 and 2000 cGy X-ray for 10 minutes, while those in the control group remained untreated. And then the pregnancy test was performed. (2) Cyclophosphamide injection. Forty 4-5-week-old male mice were divided into 3 experiment groups and a control group, the former treated with different doses of Cyclophosphamide via ip and the latter Natiichloridi Saline (N.S.) via i.p., followed by the pregnancy test. (3) Diphereline injection. Twenty 8-10-week-old male mice were equally divided into an experiment group and a control group, the former treated with Diphereline via ip and the latter N.S. via i.p., followed by the pregnancy test. (4) Identification by such pathologic examinations as TUNE1. technology, HE staining and immunohistochemical staining.</p><p><b>RESULTS</b>(1) X-ray local irradiation. The male mice of Group 1 and 2 made their female partners pregnant respectively 10 and 15 days after the X-ray irradiation, but not those of Group 3 and 4 in our 3-month observation, and those of Group 5 and 6 died respectively 2 and 5 days after the X-ray irradiation. By comparison, the controls got their female partners pregnant within 3 days after placed together. (2) Cyclophosphamide injection. The male mice of Group 1 gained weight about 7 g and achieved pregnancy 9-14 days after drug termination, those of Group 2 gained around 4 g but failed to effect pregnancy, and those in Group 3 lost weight and died respective at 3, 4 and 5 weeks during the medication, while the controls all got their female partners pregnant within 3 days after put together. (3) Diphereline injection. The 10 male mice of the experiment group effected pregnancy 3 weeks after drug termination, while the 10 controls achieved the same result with 3 days after placed together. (4) Pathologic identification: TUNEL technology showed that apoptotic cells were occasionally seen (0.71 +/- 0.12)% in the testis tissue of the control group and remarkably increased (10.36 +/- 1.48)% in the model group, with significant difference between the two groups (P < 0.05). HE staining revealed normal testis tissues and convoluted seminiferous tubules with large numbers of germ cells in the control group, but atrophied convoluted seminiferous tubules and estranged cell linkage with only Ledig's cells but no germ cells in the model group. Immunohistochemical staining showed that the positive expression rates of CD29, Hsp90alpha and CD117 were respectively (50.30 +/- 5.2)%, (41.6 +/- 3.5)% and (73.6 +/- 3.7)% in the control group, as compared with (1.3 +/- 0.2)%, 0% and (1.6 +/- 0.3)% in the model group, with significant difference (P < 0.01). The positive expression rate of p53 was (19.7 +/- 0.8)% in the control group, significantly different from that of the model group, which was (39.4 +/- 2.9)% (P < 0.01).</p><p><b>CONCLUSION</b>The animal model with sterilized testes can be made either by X-ray local irradiation of the testis or by Cyclophosphamide injection via i.p..</p>

Mesenchymal stem cells do not differentiate into "quasi-sperm" / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine whether the surviving mesenchymal stem cells (MSCs) in the testis after transplantation can differentiate into quasi-sperm.</p><p><b>METHODS</b>(1) Making an animal model with sterilized testes. Forty 4-week old white male BASB/C mice were used to establish an animal model with sterilized testes and divided randomly into an experimental and a control group. (2) Cell preparation. The MSCs from 10 gray male 129-mice were isolated, cultured and purified by Percoll density gradient centrifugation combined with the adherent method. When the MSCs grew to an adequate number, they were made into a cell suspension with NS at a concentration of 1 million cells/ml. (3) Xenogeneic transplantation of the MSCs into the testis. The MSC suspension was blindly injected into the testes of the mice in the experimental group and NS into the testes of the controls. (4) Post-transplantation observation. Forty white female BASB/C mice were adopted, each put into a box with a male mouse from the experimental group or the control group, and then observed for pregnancy.</p><p><b>RESULTS</b>In the experimental group, 8 cases of pregnancy (40%) were observed at 31-46 d (38.5 d on average), the offspring all white. In the control group, only 1 case of pregnancy (5%) was seen at 45 d, the offspring all white, too. It was suggested that the MSCs of the 129-mice failed to differentiate into functional quasi-sperm and pass their genes to their offspring, as would expectedly have been presented by a mixture of black and white. The pregnancy rates of the two groups were significantly different (P < 0.05), which indicated that MSCs could promote the healing of the testis damage.</p><p><b>CONCLUSION</b>MSCs cannot differentiate into quasi-sperm after heterogeneity transplantation into the testis, but can promote the healing of the testis damage.</p>

Isolation and culture of mouse marrow mesenchymal stem cells (MSCs) and biological characteristics of MSCs cultured in conditions for spermatogonia in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To isolate, culture and purify mouse bone marrow mesenchymal stem cells (MSCs) and observe the main biological characteristics of MSCs cultured in conditions for spermatogonia in vitro.</p><p><b>METHODS</b>The tibias and femurs were dissected from 5 - 6-week old mice and the marrow in the tibias and femurs was flushed out with medium. MSCs were isolated, cultured, purified in vitro by Percoll density gradient centrifugation combined with adherent method and identified by dynamic observation of stem cell characteristics by transmission electron microscope, HE staining, and immunohistochemical detection of cell markers. The quantities of such cytokines as IL-6, IL-8, G-CSF and SCF in culture liquid with MSCs were measured by ELISA, and compared with those of the control group. MSCs of the third generation were divided into two groups to be induced and cultured. MSCs of the control group were cultured with basal medium, while those of the experimental group with conditional medium. The results were analysed by microscopic observation, HE staining and immunohistochemical methods.</p><p><b>RESULTS</b>Pure MSCs were obtained. The cultured cells, with stem cell characteristics, shuttle-shaped at HE staining, immature under the transmission electron microscope and CD44 and CD90 positive by immunohistochemical detection, could be identified as MSCs. Compared with the control group, the quantities of IL-6, IL-8, G-CSF and SCF in the experimental group increased significantly (P < 0.05). The shapes of MSCs changed and immunohistochemical staining for CD27, CD119 and Oct-4 was positive in the experimental group, but both were just the opposite in the control group.</p><p><b>CONCLUSION</b>Pure MSCs can be obtained by Percoll density gradient centrifugation combined with adherent method and identified by dynamically observing stem cell characteristics, HE staining, observation under the transmission electron microscope and immunohistochemical detection of cell markers. MSCs can secrete cytokines such as IL-6, IL-8, G-CSF, SCF, and so on. MSCs cultured in conditions for spermatogonia may show some biological characteristics of spermatogonia.</p>

Transplantation of mouse bone marrow mesenchymal stem cells into the xenogeneic testis / 中华男科学杂志

<p><b>OBJECTIVE</b>To study transplantation of mouse bone marrow mesenchymal stem cells (MSCs) into the xenogeneic testis.</p><p><b>METHODS</b>(1) The tibias and femurs were dissected from 5-6-week-old mice. The marrow in the tibias and femurs was flushed out with medium. MSCs were isolated, cultured and purified in vitro by Percoll density gradient centrifugation combined with adherent method. (2) MSCs of the third generation were adopted and marked with Hoechest33342 for observation, and then made into cell-suspending fluid. (3) The marked MSCs were transplanted into the testis of the xenogeneic mouse by testis net injection. The biopsies of the testis tissues were carried out at different time and made into frost slices at three sites for observation.</p><p><b>RESULTS</b>(1) A lot of purified MSCs were obtained at the third generation. (2) The nucleoli of the marked MSCs showed light-yellow under the fluoroscope. (3) Xenogeneic transplantation of mouse bone marrow MSCs by testis net injection was successful, without immunoreaction. On the 1 st day after transplantation, MSCs only concentratively distributed in the medial slices, the nucleoli being light-yellow; On the 1 st and 3 rd day, MSCs dispersively distributed in the medial slices; On the 6th, 9th and 12th day, MSCs presented in all the slices of the three sites, some ranging tubally; On the 15th and 18th day, the fluorescence of MSCs weakened; On the 21 st day, the fluorescence of MSCs disappeared.</p><p><b>CONCLUSION</b>Transplantation of mouse bone marrow MSCs into the xenogeneic testis by testis net injection is effective and feasible, without immunoreaction. MSCs can survive after transplantation.</p>

The capacity of MTD method for distinguishing Mycobacterium tuberculosis and nontuberculosis mycobacteria / 中华检验医学杂志

Objective To evaluate the capacity of MTD method to distinguish between Myeobacterium tuberculosis and nontuberculosis mycobacteria.Methods Ten standard strains(including 1 H_(37)Rv strain and 9 nontuberculosis mycobacteria strains),94 clinical strains(including 48 Mycobacterium tuberculosis and 46 nontuberculosis mycobaeteria strains)and 40 sputum specimens were tested by MTD method(AMPLIFIED-MTI))and traditional methods.The results of these methods were compared.Results For all Myeobacterium tuberculosis and nontuberculosis mycobacteria strains,the agreement of MTD method and traditional method was 100%.And the positive detectable rate for sputum samples was 65% that was obviously higher than that for the direct smear(5/40),concentration smear(10/ 40)or culture(5/40).Conclusion MTD is a rapid test for identification of Mycobacterium tuberculosis and nontuberculosis mycobaeteria with high sensitivity and specificity.