Panhypopituitarism caused by a suprasellar germinoma: A case report

BACKGROUND Suprasellar germinomas are rare intracranial tumors frequently associated with permanent endocrine disorders.We present the clinical picture,treatment,and complications of suprasellar germinoma at pediatric age which,besides being lifethreatening,has lifelong endocrinological consequences.CASE SUMMARY A 12-year-old female patient was presented having had intensive headaches for three weeks and visual disturbances for six months.An ophthalmological examination revealed bilateral papilledema and a marked loss of vision.Emergency brain magnetic resonance imaging(MRI)showed a suprasellar tumor,involving the infundibulum and the optic chiasm,extending to the third ventricle.Laboratory tests confirmed decreased levels of thyroxine,cortisol,gonadotropins,and insulin-like growth factor 1.Maximal tumor reduction was performed,and immunohistopathology established the diagnosis of suprasellar germinoma.MRI of the spine and cerebrospinal fluid cytology confirmed the localized disease.Adjuvant chemotherapy and radiotherapy were performed according to the SIOP CNS GCT II protocol.A post-treatment MRI showed no residual tumor,but pituitary function had not recovered.Three and a half years after the end of the treatment,the patient is in a complete remission,requiring hormonal replacement therapy,continuous education,and psychological support.CONCLUSION This complex case highlights the importance of timely diagnosis,a multidisciplinary approach,and close follow-up in children with suprasellar germinomas.

A wheat germ-rich diet preserves bone homeostasis by regulating gut microbiota and plasma metabolites in aged rats

Bone loss caused by ageing has become one of the leading health risk factors worldwide.Wheat germ(WG)is consists of high amounts of bioactive peptides,polyunsaturated fatty acids,and dietary fibre.Currently,WG has been proven to possess strong antioxidant and anti-infl ammatory properties.We recently explored the beneficial effects and relevant mechanisms of a WG-rich diet(2.5%and 5%WG,m/m)on bone homeostasis in aged rats.Our results showed that 5%WG supplementation for 12 months effectively attenuated ageing-induced microstructural damage and differentiation activity changes in the femur.The 5%WG supplementation also signifi cantly increased the levels of total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px)(P<0.01),and superoxide dismutase(SOD)(P<0.05),and decreased infl ammatory cytokine levels(tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6))(P<0.01).Furthermore,the WG-rich diet reshaped the composition of the gut microbiota,enhancing short-chain fatty acids(SCFAs)-producing microbes and reducing infl ammation-related microbes.In addition,metabolomics analysis showed that 5%WG supplementation improved plasma metabolites related to bone metabolism.Conclusively,our study purports long-term WG-rich diet may preserve bone homeostasis by regulating gut microbiota and plasma metabolites in aged rats.

羟基脲对大鼠Sertoli-germ细胞共培养的影响

目的:观察羟基脲(Hydroxyurea,HU)对雄性大鼠Sertoli-germ细胞共培养的细胞毒性作用。方法:在Sertoli-germ细胞共培养模型中分别加入不同浓度HU(0、10-3、10-2、10-1mol/L)后,孵育96h;以及加入10-2mol/LHU后分别孵育24、48、72、96h。结果:在各给药组,96h时HU导致germ细胞从Sertoli细胞层的脱落增加,呈现量效关系。在10-3mol/L以上浓度组,细胞活性逐渐下降,细胞脱落增加。同样,随着给药时间的增加,germ细胞脱落率和乳酸脱氢酶(LDH)的漏出逐渐增加,细胞活性逐渐下降。结论:HU对雄性大鼠睾丸Sertoli-germ细胞共培养模型的germ细胞具有明显毒性作用。

使用Glo Germ开展生物安全教育初探

为提高生物技术专业本科学员生物安全基本技能,在教学过程中引入Glo Germ作为可视化教学工具,直观生动地让学员在设定的实验场景中了解模拟"病菌"的传播过程,学习污染控制技术。该举措通过师生的互动,提高了本科生的生物安全意识。

Culture of Embryonic Stem Cell by Primordial Germ Cells of Down Producing Goat

[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.

Tudor and its domains: germ cell formation from a Tudor perspective

In many metazoan species, germ cell formation requires the germ plasm, a specialized cytoplasm which often con-tains electron dense structures. Genes required for germ cell formation in Drosophila have been isolated predominantlyin screens for maternal-effect mutations. One such gene is tudor (tud); without proper tud function germ cell formationdoes not occur. Unlike other genes involved in Drosophila germ cell specification tud is dispensable for other somaticfunctions such as abdominal patterning. It is not known how TUD contributes at a molecular level to germ cell forma-tion but in tud mutants, polar granule formation is severely compromised, and mitochondrially encoded ribosomal RNAsdo not localize to the polar granule. TUD is composed of 11 repeats of the protein motif called the Tudor domain. Thereare similar proteins to TUD in the germ line of other metazoan species including mice. Probable vertebrate orthologuesof Drosophila genes involved in germ cell specification will be discussed.

用均匀设计方法优化Sertoli-germ细胞培养条件的研究

采用均匀设计方法对Ｓｅｒｔｏｌｉｇｅｒｍ细胞共培养条件进行优化。结果显示，所观察的三个因素（３或６个水平）中，培养温度和ｐＨ值对生精细胞脱落的影响最为明显，而小牛血清浓度的影响较少。回归方程提示该培养系统的适宜条件，逐步回归分析剔除了小牛血清浓度的因素，说明不必加入过多小牛血清。此外，用所获回归方程对指标值进行回算，结果提示，以脱落生精细胞数作为培养效果的观察指标比较稳定。

Antioxidant properties of wheat germ protein hydrolysates evaluated in vitro

Wheat germ protein hydrolysates were prepared by protease hydrolysis, ultrafiltration and dynamical adsorption of resin. The total amount of amino acids in 100 g wheat germ protein hydrolysates is 93.95 g. Wheat germ protein hydrolysates are primarily composed of 4 fractions: 17.78% in the relative molecular mass range of 115631512, 17.50% in 1512842, 27.38% in 842372 and 30.65% in 37276, respectively. The antioxidant properties of wheat germ protein hydrolysates were evaluated by using different antioxidant tests in vitro. 1.20 g/L wheat germ protein hydrolysates exhibit 78.75% inhibition of peroxidation in linolei acid system; and 1.6g/L wheat germ protein hydrolysates show 81.11% scavenging effect on the 1,1-diphenyl-2-picrylhrazyl radical. The reducing power of 2.50 g/L wheat germ protein hydrolysates is 0.84. Furthermore, the scavenging activity of 0.60 g/L wheat germ protein hydrolysates against superoxide radical is 75.40%; 0.50 g/L wheat germ protein hydrolysates exhibit 63.35% chelating effect on ferrous ion. These antioxidant activities of wheat germ protein hydrolsates increase with the increase of its concentration. Experimental results suggest that wheat germ protein hydrolysate is a suitable natural antioxidant rich in nutrition and nontoxic.

Study on FTIR Spectra of Corn Germs and Endosperms of Three Different Colors Combining with Cluster Analysis

[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types were studied by using Fourier transform infrared spectroscopy（FTIR） technology, combined with cluster analysis. [Result] The overall characteristics of original FTIR spectra were basically similar within the range of 700-1 800 cm^-1. The FTIR spectra were mainly composed by the absorption peaks of polysaccharides, proteins and lipids. Within the wavelength range of 700-1 800 cm^-1, there were only tiny differences in original FTIR spectra among the corn germs and endosperms of three different types. The spectra were then processed by using first derivative and second derivative. The second derivative spectra were used for hierarchical cluster analysis（HCA）. The results showed that with the wavelength range of 700-1 800 cm^-1, the second derivative spectra of the 52 samples could be better clustered according to the tree types and corn germ and corn endosperm. The clustering correct rate reached 96.1%.[Conclusion] FTIR technology, combined with cluster analysis, can be used to identify different types of corn germs and endosperms, and it is characterized by convenience and rapidness.

Improved Isolation and Culture of Embryonic Germ Cells from Guanzhong Dairy Goat

A total of 219 embryonic-germ-cell-like （EG-like） clumps were derived from 15 selected goat fetuses. Isolation of primordial germ cells （PGCs） based on co-culture with primary goat embryonic fibroblast showed no difference from traditional feeder layer-based culture method used in mouse and human. The putative primary EG colonies were multilayer clumps of compact cells with unclear cell-cell boundaries. Three subculture methods of goat EG-like colony, traditional enzymatic digestion, mechanical cutting and combination of the both, were compared in this study. As a result, EG-like colonies traditionally disassociated with collagenase 1V could be subcultured for up to 4 passages. And the mechanically disaggregated EG-like colonies were successfully maintained 9-12 passages with or without enzymatic treatment. The pluripotency of the EG-like colonies was identified by their specific marker staining, spontaneous differentiation and embryoid bodies （EBs） formation in vitro. Most goat EG-like colonies （〉 80%） were AKP positive and immunocytochemically characterized with positive SSEA-1, Oct-4 and c-kit staining but SSEA-4. Under the condition of delaying passage, goat EG-like cells could differentiate into fibroblast-like, epithelium-like, and neuron-like cells. In addition, EBs could be obtained successfully in routine hanging drop culture. The serum free culture system （feeder layer-based） used in this study was suitable for keeping PGCs and EG-like cells in their undifferentiated condition, but failed to converse them to immortal cells. These results indicated that mechanical cutting is an effective method for passaging goat EG cell colonies. However, the microenvironment of conversing EG cells to immortal cells is still unclear.

The Technology Research of Anti-oxidation Peptide Preparation by Alkaline Protease Hydrolyzing Wheat Germ Meal

Wheat germ meal is the by production of oil extracting, and a great quantity of it has been wasted, thus the quantity of lost protein is great. In order to use wheat germ meal proteins adequately, wheat germ proteins were hydrolyzed to anti-oxidation peptides by using alkaline protease. Through the single factor analysis and regression analysis, the optimized experiment conditions of hydrolysising wheat germ meal to wheat germ peptides were enzymatic quantity 0.8%（w/w）, material to liquid ratio 1∶12.3, enzymolysis time 2.1 h. Under these conditions, the scavenging effect was 49.78%,the DH was 22% and peptides content in enzymatic hydrolysate was 1.9%（w/w）.By SDS-PAGE electrophoresis,the molecular weight range of wheat germ peptides were below 10 ku and most were between 4.54 and 5.63 ku.The wheat germ proteins could be used ful y and grain resources would be saved.

Molecular and epigenetic pathogenesis of germ cell tumors

The development of germ cell tumors(GCTs)is a unique pathogenesis occurring at an early developmental stage during specification,migration or colonization of primordial germ cells(PGCs)in the genital ridge.Since driver mutations could not be identified so far,the involvement of the epigenetic machinery during the pathogenesis seems to play a crucial role.Currently,it is investigated whether epigenetic modifications occurring between the omnipotent two-cell stage and the pluripotent implanting PGCs might result in disturbances eventually leading to GCTs.Although progress in understanding epigenetic mechanisms during PGC development is ongoing,little is known about the complete picture of its involvement during GCT development and eventual classification into clinical subtypes.This review will shed light into the current knowledge of the complex epigenetic and molecular contribution during pathogenesis of GCTs by emphasizing on early developmental stages until arrival of late PGCs in the gonads.We questioned how misguided migrating and/or colonizing PGCs develop to either type Ⅰ or type Ⅱ GCTs.Additionally,we asked how pluripotency can be regulated during PGC development and which epigenetic changes contribute to GCT pathogenesis.We propose that SOX2 and SOX17 determine either embryonic stem cell-like(embryonal carcinoma)or PGC-like cell fate(seminoma).Finally,we suggest that factors secreted by the microenvironment,i.e.BMPs and BMP inhibiting molecules,dictate the fate decision of germ cell neoplasia in situ(into seminoma and embryonal carcinoma)and seminomas(into embryonal carcinoma or extraembryonic lineage),indicating an important role of the microenvironment on GCT plasticity.

Surgical treatment of metastatic germ cell cancer

Among young men between the ages of 15 and 40 years,germ cell cancer is the most common solid tumor[1].The worldwide incidence of germ cell cancer is 70000 cases.Compared to all solid tumors of men,germ cell cancer accounts for 1%of all male tumors.Nevertheless,the mortality of this rare tumor entity is about 13%since 9507 patients died worldwide of germ cell cancer.The improvement in survival of germ cell cancer patients is due to a multimodal treatment of germ cell cancer including cisplatin-based chemotherapy and surgery leading to higher cure-rates even in advanced stages[1],whereas the increasing incidence of germ cell cancers cannot be thoroughly explained.In this article we review the current indications for surgery in metastatic germ cell cancers,highlight the strength and weaknesses of techniques and indications and raise the question how to improve surgical treatment in metastatic germ cell cancer.

Bilateral Mixed Germ Cell Tumour in an Adolescent Girl with Premature Ovarian Failure

Germ cell tumours (GCTs) of the ovary are rare, comprising approximately 20% of all ovarian tumors with the malignant variant accounting for less than 5% of all ovarian neoplasms. Malignant mixed germ cell tumours are rarer still with dysgerminoma & yolk sac tumour being the most common components with three component variants being categorised in the rarest of rare varieties. Bilateral dysgerminomas occur in 15% of germ cell neoplasms but rarely present with premature ovarian failure. We present the case of an adolescent girl with a short history of abdominal pain and distension with amenorrhoea for a year. Clinical and radio ogical examination revealed a pelvic/adnexal mass with elevation of tumour markers and she had to undergo a staging laparotomy with bilateral salpingo-oophorectomy. Histopatholgy examination was reported as malignant mixed germ cell tumour of left ovary, predominantly with immature teratoma and minor components of yolk-sac tumour and dysgerminoma and right ovary with dysgerminoma. In view of mixed germ cell disease, she was planned for adjuvant chemotherapy.

苏云金芽孢杆菌芽孢萌发相关基因gerM的克隆及功能研究

依照蜡状芽孢杆菌gerM基因的保守序列设计引物,从苏云金芽孢杆菌中扩增出640bp的DNA片段。以此为探针,从苏云金芽孢杆菌部分基因组酶切文库中成功地克隆到了一个4·5kb的DNA片段。序列分析表明,该片段包含一个完整的开放阅读框,其预测的编码产物与枯草芽孢杆菌GerM蛋白具有很高的同源性,将该基因命名为gerM。RT-PCR分析表明,gerM基因仅在芽孢形成的过程中表达。通过同源重组的策略构建了gerM基因的阻断突变株。研究表明,gerM基因的破坏影响苏云金芽孢杆菌芽孢萌发的速率和比例。

Treatment Outcomes of Germ Cell Tumors of Ovary:Single Institutional Study

5%of all ovarian tumours are accounted to germ cell tumours(GCT’s).Affecting mostly young women,the highest incidence is seen in second and third decade of life.They are highly malignant but chemosensitive and more curable than their epithelial counterparts.Treating these tumors with effective surgery and combination chemotherapy survival rates have dramatically improved in recent decades.We present our experience of ovarian germ cell tumours in the department of Surgical Oncology,Rajendra Institute of Medical Sciences(RIMS),Ranchi with special emphasis on treatment outcomes.A retrospective review of hospital medical records of patients with ovarian germ cell tumours diagnosed and treated at RIMS from June 2019 to August 2020,was performed.Clinical profile and treatment outcome of patients were recorded.A total of 19 patients met criteria.The median age at diagnosis was 20 years(range 11-42 years)and all had good performance status.All except two patients underwent surgery,70.6%and 29.4%in upfront and interval debulking surgery(IDS)setting respectively.Fertility preserving surgery was done in 75%patients in the primary surgery group and 60%undergoing IDS.83.3%patients received BEP as adjuvant chemotherapy whereas 80%as neo-adjuvant chemotherapy.Majority(31.5%)patients had dysgerminoma as final histology,followed by mixed histology(26.3%),yolk sac tumour(15.7%),immature teratoma(15.7%)and choriocarcinoma(10.5%).47.3%patients were in Stage I at the time of diagnosis.78.9%patients were alive without disease,10.5%recurred,and 10.5%were lost to follow up.

Germ cell apoptosis induced by Ureaplasma urealyticum infection

Aim: To study the effect of Ureaplasma urealyticum (UU) infection on germ cell apoptosis of male rats. Meth-ods: Male rats were infected artificially with UU serotype 8 (T_(960)). Morphological changes of germ cells in the sem-iniferous tubules and the lumen of the epididymides were observed under the light microscope. Fluorescence-conjugatedpolyclonal antibodies to Fas and Fas ligand (FasL) were used to localize Fas and FasL. TUNEL staining of germ cellsand Sertoli cells was performed by the AKPase method. TUNEL-positive rate (% positive cells) and TUNEL-positivearea (area occupied by stained cells) were analysed by KS400 Image Analysis System. The DNA laddering analysiswas performed by agarose gels electrophoresis. Results: In those rats infected with UU; (1) Exfoliated germ cellswere dramatically increased. Many multinucleated giant cells were found in the seminiferous tubules and the lumen ofthe epididymides. (2) The number of TUNEL-positive cells and the TUNEL-positive area were significantly increased.(3) The expression of Fas and FasL in germ cells and Sertoli cells was up-regulated. (4) Discrete bands of fragmentedDNA were found in the testicular cells. Conclusion: In male rats, germ cell apoptosis was increased in UU infec-tion . (Asian J Androl 2001 Sep; 3: 199 - 204)

Assessment of germ cell apoptosis in cryptorchid rats

Aim: To investigate the relationship between germ cell degeneration and apoptosis in cryptorchid rats. Methods: Thirteen 21-day-old Wistar rats were made unilaterally cryptorchid by closing the left inguinal canal. At day 30 (Group 1, n=6) and day 60 (Group 2, n=7) after operation, the testes were removed for histopathological examination. The controls (n=8) were sham operated and were sacrificed at day 60. Germ cell apoptosis was assessed by means of the TUNEL method. Results: Spermatogenesis was arrested and the testicular and seminiferous tubular diameters were significantly reduced In the unilateral undescended testes (UUTs) compared with the contralateral descended testes (CDTs) and the control rats. However, atrophic changes, pathological calcification, necrosis of seminiferous tubule, and absence or sloughing of germ cells were not found in all the animals. The spermatocytes were the main type of germ cells undergoing apoptosis in all the groups. In the UUTs, there was a significant and time-dependent increase in the mean apoptotic index. By 60 days after surgery, increased apoptosis in germ cells was also observed in the CDTs. Conclusion: Apoptosis is the predominant mechanism of germ cell death rather than atrophy and necrosis in cryptorchidism.

Generation of male germ cells from induced pluripotent stem cells （iPS cells）： an in vitro and in vivo study

Recent studies have reported that induced pluripotent stem （iPS） cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not known. The objective of this study was to investigate the differentiation potential of mouse iPS cells into spermatogonial stem cells and late-stage male germ cells. We used an approach that combines in vitrodifferentiation and in vivotransplantation. Embryoid bodies （EBs） were obtained from iPS cells using leukaemia inhibitor factor （LIF）-free medium. Quantitative PCR revealed a decrease in Oct4 expression and an increase in StraSand Vasa mRNA in the EBs derived from iPS cells, iPS cell-derived EBs were induced by retinoic acid to differentiate into spermatogonial stem cells （SSCs）, as evidenced by their expression of VASA, as well as CDH1 and GFRal, which are markers of SSCs. Furthermore, these germ cells derived from iPS cells were transplanted into recipient testes of mice that had been pre-treated with busulfan. Notably, iPS cell-derived SSCs were able to differentiate into male germ cells ranging from spermatogonia to round spermatids, as shown by VASA and SCP3 expression. This study demonstrates that iPS cells have the potential to differentiate into late-stage male germ cells. The derivation of male germ cells from iPS cells has potential applications in the treatment of male infertility and provides a model for uncovering the molecular mechanisms underlying male germ cell development.

Expression of germ cell nuclear factor in mouse germ cells and sperm during postnatal period

Aim: To assess the spatial and temporal expression of germ cell nuclear factor (GCNF) in male mouse germ cells during postnatal development and in sperm before and after capacitation. Methods: The indirect immun-ofluorescence method with anti-GCNF antiserum was used to investigate the GCNF expression in mice at day 8, 10, 14, 17, 20, 28, 35, 70, and 420 after birth and in sperm before and after capacitation. Results: With the proceeding of spermatogenesis, GCNF was first detected in the nuclei of spermatogonia and a few early stage primary sperma-tocytes at day 8, which was increased gradually at day 10 to 14 inclusive. From day 17 to day 20, the GCNF was concentrated in round spermatids, while both spermatogonia and early stage primary spermatocytes became GCNF negative. From day 28 until day 420, strong GCNF expression was shown in round spermatids and pachytene spermatocytes, while spermatogonia, early primary spermatocytes and elongating spermatids were all GCNF negative. In addition, it was also found that GCNF was localized on the acrosomal cap region of spermatozoa and there was a big change in GCNF expression during capacitation, from 98 % GCNF positive before capacitation to about 20 % positive following capacitation. The localization of GCNF in caput and cauda spermatozoa was similar. Conclusion: GCNF may play important roles in spermatogenesis, capacitation and fertilization.