Clinicopathological analysis of 19 cases with congenital nonprogressive hemangiomas / 临床与实验病理学杂志

Purpose To investigate the clinicopathological features and differential diagnosis of congenital nonprogressive hemangiomas in children.Methods The clinic pathologic data and follow-up results of 19 cases of congenital nonprogressive hemangiomas were collected.The histopathologic features of congenital nonprogressive hamangiomas were observed with HE staining and immunohistochemistry of EnVision two-step.Results Of the 19 patients,13 were males and 6 were females,with an average age of 52.4 months and medium age of 48 months.9 cases occurred in the limbs and trunk,10 lesions were located in head and face included 1 case with multifocal lesions.Grossly,the specimens were irregular solid mass with range from 0.4 cm to 7 cm in maximum diameter.Microscopically,the lesions were composed of capillary lobules that separated by abnormal dense fibrotic dermal stroma typically containing a prominent component of large vessels.Immunohistochemically,the tumor cells were positive for CD31,CD34,and WT1,but negative for D2-40 and GLUT1.All eases underwent surgical resection,and the follow-up period was 1-24 months.There was no recurrence or metastasis.Conclusion Congenital nonprogressive hemangioma is a rare benign vascular tumor in children,with characteristic clinical and pathologic features.So the exact diagnosis is very important for both pathologists and clinicians.

Detection of DPY19L2 gene mutation in a globozoospermia patient / 中华男科学杂志

<p><b>OBJECTIVE</b>Globozoospermia is mostly associated with homozygous deletion of the DPY19L2 gene. This study aimed to investigate the DPY19L2 gene mutation in a globozoospermia patient.</p><p><b>METHODS</b>We observed the sperm histomorphology of a patient with globozoospermia using Wright-Giemsa's staining and transmission electron microscopy, detected the mutation of the DPY19L2 gene by PCR amplification and DNA sequencing, and compared the findings with the sequences issued in the Genbank.</p><p><b>RESULTS</b>Wright-Giemsa's staining showed that all the spermatozoa were round-headed and lacked the acrosome, with the head nucleus darkly, fully and densely stained. Transmission electron microscopy revealed larger round sperm heads, with an even layer of unit membrane surrounding the nuclei and dispersed cytoplasmic vacuoles but no acrosomal structure. No DPY19L2 gene mutation was found by PCR amplification and DNA sequencing.</p><p><b>CONCLUSION</b>No homozygous mutation of the DPY19L2 gene was found in the globozoospermia patient, and therefore some other disease-causing genes might be involved.</p>

DPY19L2 gene and globozoospermia: an update / 中华男科学杂志

Globozoospermia is a severe teratozoospermia, and the cases with 100% round-headed sperm are rare clinically. Globozoospermia is generally characterized by absence or abnormality of acrosome, accompanied by round-headed sperm with deranged midpiece and tails. The acrosome normally contains the enzymes that enable sperm to fertilize oocytes, while defective sperm cannot independently fertilize oocytes either in vivo or in vitro, and therefore globozoospermia makes males infertile clinically. Recent studies show that the deletion of the DPY19L2 (dpy-19-like 2) gene is a major cause of globozoospermia. This paper updates the relationship between DPY19L2 and globozoospermia to provide some evidence for further studies on the gene diagnosis and molecular mechanisms of globozoospermia.

High efficiency of L-glutamine production by coupling genetic engineered bacterial glutamine synthetase with yeast alcoholic fermentation system / 生物工程学报

Glutamine is an important conditionally necessary amino acid in human body. The effort is to establish a new and high efficient L-glutamine production system instead of traditional fermentaion. In this paper, high efficiency of L-glutamine production is obtained by coupling genetic engineered bacterial glutamine synthetase (GS) with yeast alcoholic fermentation system. Glutamine Synthetase gene (glnA) was amplified from Bacillus subtilis genomic DNA with primers designed according to sequences reported in EMBL data bank, then it was inserted into expression vector PET28b, the sequence of glnA was proved to be the same as that reported in the data bank by DNA sequencing. After transformation of this recombinant plasmid PET28b-glnA into BL-21 (DE3) strain, Lactose and IPTG were used to induce GS expression at 37 degrees C separately. Both of them can induce GS expression efficiently. The induced protein is proved to be soluble and occupies about 80% of the total proteins by SDS-PAGE analysis. The soluble GS was purified by Ni2+ chelating sepharose colum. After purification, the purified enzyme was proved active. Results reveal that the optmum temperature of this enzyme is 60 degrees C and optmum pH is 6.5 in biosynthetic reaction by using glutamate, ammonium choloride and ATP as substrates. After induction, the enzyme activity in crude extract of BL-21/PET28b-glnA is 83 times higher than that of original BL-21 extract. Mn2+ can obviously increase the activity and stability of this enzyme. Experiments show that the transformation efficiency of glutamate to glutamine is more than 95%. Because of the high cost from ATP, a system coupling GS with yeast for ATP regenaration was established. In this system, GS utilizes ATP released by yeast fermentation to synthesize L-glutamine. Yeast was treated by 2% toluence to increase its permeability and a yeast named YC001 with high yield of glutamine by coupling with recombinant GS was obtained. The good efficiency was achieved with the presence of 250 mmol/L glucose and 200 mmol/L phosphate, the transformation efficiency of glutamate to glutamine in this system is more than 80%, the average yield of glutamine is about 22g/L. This provides the basis for future large scale production of L-glutamine.