There remains a significant gap in our quantitative understanding of crosstalk between apoptosis and necroptosis pathways. By employing the SWATH-MS technique, we quantified absolute amounts of up to thousands of proteins in dynamic assembling/de-assembling of TNF signaling complexes. Combining SWATH-MS-based network modeling and experimental validation, we found that when RIP1 level is below ~1000 molecules/cell (mpc), the cell solely undergoes TRADD-dependent apoptosis. When RIP1 is above ~1000 mpc, pro-caspase-8 and RIP3 are recruited to necrosome respectively with linear and nonlinear dependence on RIP1 amount, which well explains the co-occurrence of apoptosis and necroptosis and the paradoxical observations that RIP1 is required for necroptosis but its increase down-regulates necroptosis. Higher amount of RIP1 (>~46,000 mpc) suppresses apoptosis, leading to necroptosis alone. The relation between RIP1 level and occurrence of necroptosis or total cell death is biphasic. Our study provides a resource for encoding the complexity of TNF signaling and a quantitative picture how distinct dynamic interplay among proteins function as basis sets in signaling complexes, enabling RIP1 to play diverse roles in governing cell fate decisions.
Animals ; Apoptosis ; Caspase 8/metabolism* ; GTPase-Activating Proteins/metabolism* ; HEK293 Cells ; Humans ; Mice ; Mice, Knockout ; Necroptosis ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism*

Objective To investigate the clinical characteristics,antibiotic susceptibilities and serotypes of Group B Streptococcus(GBS)isolated from neonatal meningitis to provide references for the prevention and treatment of neonatal GBS meningitis. Methods From June 2013 to June 2016,we surveyed the GBS strains iso-lated from purulent meningitis of < 90 days infants from Guangzhou Women and Children′s Medical Center. The GBS isolates were identified and the minimum inhibitory concentration of the antibiotics was determined by Vitek 2 Compact automatic bacterial identification system.GBS serotyping was performed using Strep-B-Latex?rapid latex agglutination test kit. Results A total of 46 cases of neonatal GBS meningitis,15 cases of early-onset infection and 31 cases of late-onset infection were diagnosed. 78.3% of GBS meningitis with varying degrees of complica-tions.Among 41 survivors with 3~24 months follow-up,50% of the early-onset and 44.8% of the late-onset GBS meningitis with varying degrees of neurological sequelae.Four capsular types were identified among the 46 isolates, serotype-Ⅲwas the most prevalent(73.9%),followed by Ib(19.6%),V(4.3%)and Ia(2.2%).All the isolates were susceptible to penicillins,cephalosporins,linezolid and vancomycin. Conclusion The highly pathogenic serotype-Ⅲ was the predominant serotype among neonatal GBS meningitis in Guangzhou,Therefore,it is neces-sary to strengthen the epidemiological surveillance of GBS invasive infection and the effective implementation of pre-ventive measures.

OBJECTIVETo analyze the karyotypes and SRD5A2 gene mutations in 25 patients with sporadic or familial hypospadias.

METHODSThe patients included 10 adults and 15 children, whose chromosomes were analyzed by G-banded karyotyping, and the SRD5A2 genes were sequenced.

RESULTSTwo patients were found to have an abnormal karyotype, while eight have carried compound heterozygous mutations of the SRD5A2 gene, which included 5 genotypes formed by 6 types of mutations, i.e., p.G203S/p.R227Q, p.R227Q/p.R246Q, p.Q6X/p.Q71X, p.L20P/p.G203S, and p.Q71X/p.R227Q. Mutations of the SRD5A2 gene were present in 32% (8/25) of all patients, 35% (8/23) in those with a normal karyotype, and 44.4% (8/18) in those with proximal type hypospadia. Bioinformatic analysis, literature review and pedigree analysis confirmed that all such mutations are pathogenic.

CONCLUSIONChromosomal anomalies and mutations of the SRD5A2 gene are the main cause of hypospadias. Sequencing of the SRD5A2 gene may explain the etiology of nearly half of the patients with proximal type of hypospadas but a normal karyotype, which can facilitate genetic consulting.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase ; genetics ; metabolism ; Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; Female ; Humans ; Hypospadias ; enzymology ; genetics ; Infant ; Infant, Newborn ; Karyotyping ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutation ; Young Adult

Objective To observe the ultrastructure of blood-brain barrier in the nude mouse model of brain me-tastases from lung cancer by transmission electron microscopy using lanthanum nitrate tracing.Methods PC-9 cells (1 × 106/0.1 mL) in logarithmic phase were respectively injected into six nude mice ( model group) selected from eight nude mice randomly via the left ventricle, the other two mice without any treatment as the control group.The general status of the mice was observed after implantation.In the fourth week all the mice were sacrificed and brain tissue samples were taken and prepared for transmission electron microscopic observation using lanthanum nitrate tracing.besides, the lung and brain were removed and stained with HE to detect the presence of tumor metastasis.Results Mice in the model group began to lose weight almost simultaneously in the third week and became moribund slowly, and were all sacrificed at the fourth week when showing clear signs of cachexia.At autopsy, the thoraxes were clear, with normal lungs.Histology showed evidence of brain metastasis in all the six mice.The electron microscopy showed that lathanum nitrate tracer was escaped from the capillaries and diffusely or sparsely distributed in the brain tissues of the model group mice, however lathanum nitrate tracer was still confined in the capillary lumen in the mice of control group.Conclusions The diffuse lathanum nitrate tracer in the brain parenchymal tissue indicates the impairment of blood-brain barrier in the nude mouse model of lung cancer brain metastasis and the formation of these metastases is accompanied with the destruction of blood brain barrier.

Objective To investigate the distribution of 39 kinds of human papillomavirus(HPV)infection genotypes in female cervical cells and its clinical significance.Methods 39 types of HPV DNA were extracted from 434 samples of female cervical cells. The gene amplification combined with the gene chip technique was adopted to detect 39 kinds of HPV genotype.And the clinical da-ta of the patients were analyzed.Results Among 434 samples of female cervical cell,175 cases were HPV positive,the total HPV infection rate was 40.32%(175/434).Among them,105 cases were the single type HPV infection with the positive detection rate of 24.19%(105/434)and 70 cases were the multiple types HPV infection with the positive detection rate of 16.13%(70/434).Among single type HPV infection,31 cases were the HPV18 infection with the positive detection rate of 17.71%(31/175),which was the main HPV infection type;followed by HPV16 in 12 cases with the positive detection rate of 6.86%(12/175)and HPV52 in 11 cases with the positive detection rate of 6.29%(11/175).Among the multi-type HPV infection,each 2 cases were HPV 6+54,HPV 18+52,HPV 51+68 infection respectively,each accounted for 2.86% of the multi-type HPV infection,which were the main infection types.Conclusion HPV 16,18,52 and HPV 6+54,HPV 18 +52 and HPV 51 +68 are the main HPV infection genotypes of fe-male cervical cells.The gene amplification combined with the gene chips technique is a method suitable for clinically conducting the HPV genotyping diagnosis and the molecular epidemiologic research of HPV infection.Along with the increase of detected HPV genotypes,the HPV infection rate is also increased,its genotypes combinations trend towards diversification.

Laboratory of electron microscopy is representative of large instruments laboratories in medical research.And the operation and management in this kind of laboratory are different.Case of forty years of operation and management in the medical laboratory of electron microscopy,this paper analyzed and summarized its successful experiences in the research management system,personnel system and the aspect of equipment maintenance management.Provide a management reference for medical research institutions with the laboratory of similar large-scale instruments.

Objective To identify the typical imaging features of steatocystoma multiplex on mammography.Methods Mammographic findings in 9 patients with clinical and pathological diagnosis of steatocystoma multiplex were analyzed along with a review of the current literature.Results Four of the nine patients with steatocystoma multiplex had a positive family history.Nine patients showed multiple,round,thin-walled fatty,radiolucent nodules with well-defined margin.These nodules are located in the superficial layer of the axilla bilaterally.They are also seen on the skin of breast(7 cases),the anterior chest wall (4 cases) and the upper arm (3 cases).Conclusion Recognition of the characteristic mammographic features of Steatocystoma multiplex is important in the management of these patients.

OBJECTIVE: We used the SD rat's bone marrow stromal cells (BMSCs) cultured in vitro to observe the effects of Bugu Mixture on the apoptosis and to explore the molecular biologic mechanism of the treatment of osteoporosis with Bugu Mixture. METHODS: BMSCs were separated from the bones of the extremities of SD rats in vitro. The morphologic changes, the apoptosis cell cycles, the mitochondrion membrane potential changes, and the Bcl-2 and Bax gene expression were observed, and the effects of Bugu Mixture on the course of cell apoptosis were evaluated. RESULTS: The earlier use of Bugu Mixture could decrease the cells blocked in G0/G1 phase, and promote their synthesis of DNA in S phase. The expression of Bcl-2 was higher in the Bugu Mixture group than that in the all-trans retinoic acid (ATRA) induced group, and the expression of Bax was lower in the Bugu Mixture group than that in the ATRA induced group. The mitochondrion membrane potential descended significantly in the Bugu Mixture group than that in the ATRA induced group. CONCLUSION: The mechanism of the treatment of osteoporosis with Bugu Mixture is that the earlier use of Bugu Mixture can decrease the amount of apoptostic cells induced by ATRA, thus promoting the cell mitosis and restraining the apoptosis. It can also act as a protector to Bcl-2 located on the mitochondrion membrane. By preventing the transferring of the Bax protein from cell-plasma to mitochondrion membrane, it takes the advantage of Bcl-2 in forming Bcl-2/Bax homodimer so as to prevent the opening of the permeability transition pore to avoid the changing of mitochondrion membrane potential and the destruction of biosynthesis caused by the mitochondrion release of apoptosis inducing factors and to reach the objective of restraining apoptosis.

OBJECTIVETo observe Trichomonas vaginalis (T. vaginalis) adhering to and phagocytizing male genitourinary epithelial cells in order to study the pathogenetic mechanism of male trichomoniasis.

METHODSCultured T. vaginalis bodies were incubated with male genitourinary epithelial cells, and then the ultrastructure was observed with transmission electron microscopy.

RESULTST. vaginalis adhered to epithelial cells like amoeba, and formed pseudopodium or surface invagination surrounding or nibbling other parts of the epithelial cytoplasm.

CONCLUSIONST. vaginalis has the speciality of adhering to and phagocytizing to male genitourinary epithelial cells. Genitourinary epithelial cells may be injured directly by the phagocytosis of T. vaginalis. Attention has to be paid to the correlation.

Animals ; Bacterial Adhesion ; Epithelial Cells ; parasitology ; Genitalia, Male ; parasitology ; Humans ; Male ; Microscopy, Electron ; Trichomonas vaginalis ; ultrastructure

Objective To study the ultrastructure of neural stem cell neurosphere cultured in vitro. Methods Neural stem cell neurospheres from the rat brain were observed under transmission electron microscope or with stain of lanthanum nitrate, ruthenium red and tannic acid. Results The conjunction between the cells within neurosphere was loose and no tight junction was observed. Neural stem cells were proliferating lively. Some neural stem cells differentiated into cells with process and structure of axon, even showed the structures similar to myelin.Conclusion The ultrastructure of neural stem cell neurosphere cultured in vitro was revealed.