Electroencephalography ; Epilepsies, Myoclonic ; diagnosis ; therapy ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Prognosis ; Spasms, Infantile ; diagnosis ; Time Factors

Adult ; Brain Neoplasms ; metabolism ; pathology ; surgery ; Female ; Ganglioglioma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

OBJECTIVETo construct and identify a lentiviral vector of RNA interference targeting human Notch-1 gene.

METHODSTo determine the Notch-1 gene sequences, three RNAi target sequences (shRNA1-3) were designed in accordance with the RNAi sequence design principles and cloned into the lentiviral vector pLenOR-THM by endonuclease BamH I restriction, EcoR I double digestion, and T4 DNA-ligase ligation. After the transformation into competent DH5alpha bacteria, the candidate clones were identified by Kpn I and EcoR I double digestion and DNA sequencing. The recombinant and three packaging plasmids were co-transfected into human embryonic kidney cell line 293T cells by lipofectamine to produce the lentiviral particles. The viral titer was determined. The 293T cells were infected by the lentiviral particles obtained, and transfection efficiency was assessed using a fluorescent microscope. The lentiviral vector particles were also transfected into ACC-M cells. The Notch-1 expression in the transfected cells was assayed by quantitative reverse transcription polymerase chain reaction (QRT-PCR) and Western blot analysis.

RESULTSThe lentiviral RNAi vector pLenOR-THM-Notchl for Notch-1 gene was constructed successfully. Strong green fluorescence was observed in the 293T cells under fluorescent microscope after co-transfection of the cells with the four plasmids of lentiviral vector. The virus in the supernatant reached a titer of 5.8 x 10(8) TU x mL(-1). The transfection efficiency of the collected virus exceeded 90% in 293T cells with 1 as a multiplicity of infection. The third lentiviral vector was found to significantly inhibit the Notch-1 expression at the mRNA and protein levels.

CONCLUSIONThe lentiviral RNAi vector of Notch-1 has been successfully constructed and identified.

Cell Line ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Notch1 ; Transfection

It is to observe the therapeutic action of Guizhi Fuling capsule and the combination of active ingredients on model rats with uterine leiomyoma. The hysteromyoma rats models was established in rats by loading eatrogen, to observe the effect on pathological condition of uterus, uterus wet weight, the content of estradiol and progesterone. Guizhi Fuling capsule and the combination of active ingredients remarkably decreased uterus weight, restrained the excess proliferation of the smooth muscle of uterus, decreased the estraiol and progesterone in blood serum. Guizhi Fuling capsule and the combination of active ingredients can restrain the formation of hysteromyoma in a dose-dependent manner. Perhaps the combination of active ingredients is the material foundation of antihysteromyoma.
Animals ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Estradiol ; blood ; Female ; Leiomyoma ; blood ; drug therapy ; pathology ; Progesterone ; blood ; Rats ; Rats, Wistar ; Uterine Neoplasms ; blood ; drug therapy ; pathology

Objective To observe the effect of Ti-6AL-4V particles on morphological and func- tional changes of osteoclast in vitro.Methods Mature osteoclasts separated from New Zealand Rabbits were cultured on glass slices and cortical bone slices.The experimental group was stimulated by Ti-6AL- 4V particles at concentration of 0.1 mg/ml.The cells were stained with TRAP at different culture time to observe the morphological variety.The bone resorption pits on bone slices were stained by toluidine blue and the resorption areas analyzed by computer image analysis software.Results Osteoclasts phagocy- tosed the particles,with irregular shapes,deeper TRAP stain and earlier apoptosis.Stimulation by Ti- 6AL-4V particles brought about larger area of bone absorption lacuna.Conclusion Osteoclasts have the ability to phagocytose Ti-6AL-4V particles,which leads to morphological and functional changes and enhances bone resorption.

Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P＜0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P＞0.05),but these parameters significantly changed before and after HVHF (P＜0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P＜0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.

Objective Through detecting the expression of erbB-2 and erbB-3 of epithelial growth factor receptor family in clinical specimens of salivary adenoid cystic carcinoma, to explore the role of erbB-2 and erbB-3 in the carcinogenesis and development of salivary adenoid cystic carcinoma. Methods Thirty-four specimens of adenoid cystic carcinoma were tested with immunohistochemical assay. Then the relationship between the expressions of erbB-2, erbB-3 and the gender, age of patients, TNM staging, tumor size and nerve invasion were analyzed.Results The positive expression ratios of erbB-2 and erbB-3 were 64. 7% and 32.4% respectively. And 29.4% of tumors were positively stained both by erbB-2 and erbB-3. The expression of erbB-3 was significantly correlated with TNM staging of tumors (P = 0. 026). The correlationship between the expressions of erbB-2 and erbB-3 was also found. Conclusions erbB-3 and erbB-2 may play important roles in the carcinogenesis and development of salivary adenoid cystic carcinoma. The positive expression of erbB-3 was associated with the biological behavior of tumors and prognosis of patients, and might become one of markers to evaluate the prognosis of patients.

Objective To detection the urine of bacteria hyphae and intracellular bacterial communities in patients with indwelling urinary catheter and discuss intracellular bacterial comnmunities in the pathogenesis of catheter-related urinary tract infection.Methods From May 2014 to February 2016,95 cases with D-J stent indwelling were enrolled in this study,including 38 male patients and 57 female patients.The mean age was (43 ±21)years old,ranging from 25 to 83 years old.We recorded those patient g clinical symptoms,middle urine culture results.If the middle urine culture was positive,further pathology test and scanning electron microscopy for bacteria hyphae and intracellular bacterial communities would be considered.Results The middle urine culture showed positive in 21 cases (22％,21/95);The classification of bacteria included E.coli in 11 cases,dung enterococcus in 2 cases,klebsiella pneumonia in 4 cases,pseudomonas aeruginosa in 3 cases,epidermis staphylococcus aureus in 1 case.Among those 21 patients,9 cases had the symptoms of fever and shiver.Urine pathology testing found hyphae in 6 cases (6％,6/95).all others were E.coli infection.For scanning electron microscope,6 cases were found rodshaped bacteria and hyphae.3 cases were found intracellular bacterial communities.Conclusions The presence of intracellular bacterial communities made urothelial itself the source of endogenous bacteria of urinary tract infection.Catheter-related urinary tract infections in patients with recurrence maybe basically homology bacteria.

OBJECTIVEThis study aimed at finding sensitive indicators in diagnosing facial growth pattern by studying the relationship between the indicators which can reflect facial growth pattern according to the previous studies and posterior-anterior ratio.

METHODSThe cephalometric radiographs of 61 teenage children were measured, the posterior-anterior ratio and the other 12 indicators including Y-axis angle, facial axis angle, cranial basal angle, and the four angles (The sagittal horizontal planes were analyzed using statistic methods.

RESULTSThe MP-SN angle was significantly coincided with the posterior-anterior ratio and Pearson coefficient is -0.922, and little correlation was found between the angles of four sagital horizontal planes and the posterior-anterior ratio. The positive correlation was found between the length of mandibular ramus and posterior-anterior ratio.

CONCLUSIONThe sensitive indicators for confirming facial growth pattern are the posterior-anterior ratio and the MP-SN angle.

Adolescent ; Cephalometry ; Child ; Dental Occlusion ; Face ; anatomy & histology ; Facial Bones ; anatomy & histology ; growth & development ; Female ; Humans ; Male ; Mandible ; anatomy & histology ; growth & development ; Maxillofacial Development ; Reference Values

Objective To investigate the prognostic factors of cervical high-grade squamous intraepithelial lesions treated by cold knife conization with negative margin.Methods Two hundred and sixty-six women with cervical high-grade squamous intraepithelial lesions treated by cold-knife conization with negative margins at Beijing Hospital between Jan 1999 and Jan 2004 were analyzed retrospectively.All patients were followed up with cytology,high-risk human papillomavirus(HPV)test and eolposcopy if necessary.Results The cervical CIN recurrence rate was 8.6% with no incidence of invasive cervical cancer after a median follow-up of 46 months.The recurrence was related to the grade of lesions and gland involvement pathologically.One of 20(5.0%)cases with cervical intraepithelial neoplasia(CIN)Ⅱ,9 of 164(5.5%)cases with CIN Ⅲ(excluding carcinoma in situ,CIS)and 13 of 82(15.8%)cases with CIS recurred(P