OBJECTIVETo investigate the effect of RNA interference by targeting human telomerase reverse transcriptase (hTERT) mRNA in the larynx cancer cell line, Hep-2.

METHODSThe primary structures of hTERT cDNA were found in GenBank. Then the structure analysis were done according to RNAi strategy which determined the specific base sequences to design shRNA plasmid. Two types of plasmid, pshRNA1 and pshRNA2, involved in fluorescein gene were synthesized based on the specific base sequences. Control pshRNA3, a random sequence, and control pshRNA4, without additional specific sequence were also constructed. Cells were treated daily with pshRNA1-4 or normal culture medium respectively. The pshRNA1-3 was identified by electrophoresis. After administration of pshRNA1-4, fluorescence expression was detected by confocal microscopy, the expression of hTERT of the transfected cells was determined by Western blotting, telomerase activity was measured by TRAP-PCR ELISA, cell viability was determined by MTT assay, morphological changes and apoptosis were examined by inverted microscope and TUNEL respectively.

RESULTSThere was a 400 bp balteum in pshRNA1-3 after cut by SalI, which was identical with the size of the objective gene. Many cells presented green fluorescence after being treated by pshRNA1-4, but there are much more dead green fluorescent cells in the pshRNA1 and pshRNA2 group. hTERT protein and telomerase activity was significantly decreased after treated by pshRNA1 or pshRNA2. It was observed that treatment with pshRNA1 or pshRNA2 in the presence of a valid transfection reagent could reduce cell viability of Hep-2 cells within 96 h (P < 0.01). Under the same culture conditions, cells grew more sparsely and the number of apoptotic cell increased significantly.

CONCLUSIONSshRNA plasmid directed against human telomerase reverse transcriptase can effectively transfect Hep-2 cells. shRNA targeted hTERT gene can significantly inhibit the growth and proliferation of Hep-2 cells, which results in apoptotic cell death. RNA interference may be a promising strategy for the treatment of laryngeal cancer.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Plasmids ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; Telomerase ; biosynthesis ; genetics ; Transfection

OBJECTIVETo provide the experience for early diagnosis and management of facial nerve neuromas, and to discuss the clinic and imaging feature of facial nerve schwannoma and facial nerve fibroma in 22 cases.

METHODSTwenty cases facial nerve schwannoma and two cases of facial nerve neurofibroma were diagnosed and reviewed retrospectively. Surgical removal were performed through the middle cranial fossa in 2 cases, through intratemporal approach in 8 cases, through intraparotid approach in 2 cases, and combined intra-temporal with out-temporal approaches in 10 cases. Seventeen cases underwent facial nerve graft for repairing a facial nerve defect. Great auricular nerve was used in 3 cases with intratemporal approach and 1 case with intratemporal combined intraparotid approach. Sural nerve graft was used in 5 cases with intratemporal approach and 8 cases with intra-temporal combined intraparotid approach. Two cases were employed two-stage facial muscle flap-plasty.

RESULTSFacial nerve neuromas were totally removed in 21 cases and subtotal neuroma removed in 1 case. In these cases, 20 patients were no recurrence and 1 patient was lost follow-up. One patient with subtotal neuroma removal received Gamma Knife treatment before and after surgery, and this case was no recurrence. The CT imaging of the temporal bone showed that schwannoma was separated "white mass" with smooth margin along the region of facial nerve without intact canal. But neurofibroma locate in enlarge fallopian with intact canal. Magnetic resonance imaging had the advantage of evaluating all segments of the facial nerve and showed continuity of intratemporal and intraparotid mass with the facial nerve. Pathological results indicated that 20 cases were diagnosed as facial nerve schwannoma and 2 cases were neurofibroma.

CONCLUSIONSAlthough tumors originating from the facial nerve are extremely rare, it is possible to make early diagnosis through finding clinical feature and imaging methods. Generally, systematic surgical approach for tumor removal and facial nerve reconstruction should be considered in the cases with facial neurinoma.

Adolescent ; Adult ; Child ; Cranial Nerve Neoplasms ; diagnosis ; pathology ; surgery ; Facial Nerve ; pathology ; transplantation ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Neurilemmoma ; diagnosis ; pathology ; surgery ; Neurofibroma ; diagnosis ; pathology ; surgery ; Neuroma ; diagnosis ; pathology ; surgery ; Retrospective Studies ; Young Adult

The study was aimed to investigate the potential immunotherapeutical values of umbilical cord tissue-derived mesenchymal stem cells (UC-MSC) on patients with chronic idiopathic thrombocytopenic purpura (ITP). UC-MSC was cocultured in vitro with splenocytes isolated from ITP patients who experienced splenectomy. The level of IgG antiplatelet antibody (PAIgG) was determined by a competitive micro-enzyme-linked immunosorbent assay (ELISA) method. The proliferation of platelet-reactive CD4+ T lymphocytes was also measured in the presence of UC-MSCs. The results showed that UC-MSCs could stimulate the spontaneous secretion of PAIgG in supernatants; In the platelet-inducing condition, UC-MSC inhibited the production of PAIgG at a low ratio of 1 UC-MSC to 100 splenocytes, but promoted at a high proportion of 1 UC-MSC to 10 splenocytes. Moreover, UC-MSC exerted a suppressive effect on proliferation of platelet-reactive T helper cells in a dose-dependent manner. It is concluded that the UC-MSCs can regulate secretion of antiplatelet antibodies in vitro. Its concrete regulation mechanism and potential immunotherapeutical value are need to further study.
Antibodies ; metabolism ; Blood Platelets ; immunology ; CD4-Positive T-Lymphocytes ; cytology ; Cell Proliferation ; Humans ; Infant, Newborn ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; physiology ; Purpura, Thrombocytopenic, Idiopathic ; metabolism ; Spleen ; cytology ; Umbilical Cord ; physiology

OBJECTIVETo prepare pneumolysin as a new protein carrier of vaccine against otitis media with genetic engineering technology and establish the base of the study on pneumococcal conjugative vaccines.

METHODSGenomic DNA was isolated from streptococcus pneumoniae. A pair of primers which included two restriction sites was designed based on the published pneumolysin gene sequence. The pneumolysin gene was amplified from pneumococcal DNA with PCR technology. The restriction enzyme digested fragment was linked into the cloning vector PET-28a and the recombinant plasmid DNA containing pneumolysin was then transfected into host cell E. coli JM109 (DE3).

RESULTSDNA fragments were subcloned to construct the complete pneumolysin gene by a conventional coning and PCR. The inserted pneumolysin gene sequence was confirmed by DNA sequencing and the pneumolysin protein was successfully expressed. The relative molecular mass of the expressed product was 52 000. The expressed product amounted to 8% of the total host cell protein.

CONCLUSIONSThe pneumolysin gene was successfully cloned into host cell using genetic engineering technology. The recombinant pneumolysin was expressed and purified for preparation. This work laid a foundation of the preparation of pneumococcal conjugative vaccines.

Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Genetic Engineering ; Genetic Vectors ; Plasmids ; Pneumococcal Vaccines ; genetics ; Streptococcus pneumoniae ; genetics ; Streptolysins ; biosynthesis ; genetics

OBJECTIVETo describe the epidemiological characteristics of overweight and obesity among people aged 18 and over in Guangdong province in 2002, and to identify the populations and regions under high risk.

METHODSCross-sectional survey was used through sampling on multi-stage randomized clusters. Data of socialeconomic status were collected by face-to-face interview. Data on weight and height was obtained through physical check-ups.

RESULTSA sample size of 15 130 people and the mean body mass index (BMI) was 22.03 +/- 3.38 with no significant difference between males and females. However the significant difference was seen between cities and rural areas. The crude prevalence rate of overweight and obesity were 16.8% and 1.8%, and the age-adjusted rate were 15.0% and 1.7%, respectively. The crude rate of overweight in cities (24.8%) and males (17.5%) were higher than that in rural areas (9.4%) and females (16.2%). The crude rate of obesity in cities was seen higher than that in the rural areas, but not significantly different between females and males. The results of logistic regression analysis showed that the major risk factors influencing overweight would include household income, age, gender, smoking habits, physical exercises and location of residence.

CONCLUSIONAbout one sixth of the citizens in Guangdong province were considered to be overweighted and obesive had become an important public health problem. Integrated measures must be taken for prevention and control.

Adolescent ; Adult ; Body Mass Index ; China ; epidemiology ; Cross-Sectional Studies ; Diet ; Exercise ; Female ; Humans ; Male ; Obesity ; epidemiology ; Overweight ; Prevalence ; Risk Factors ; Sampling Studies ; Socioeconomic Factors ; Surveys and Questionnaires

OBJECTIVETo study the expression of specific anti- platelet glycoprotein autoantibodies GP II b/III a, GP I b/IX and GP I a/II a in primary immune thrombocytopenia (ITP), and to evaluate the relationship between the therapeutic effect and the expression of specific anti- platelet glycoprotein antibodies GPIIb/IIIa, GPIb/IX and GPIa/IIa.

METHODSAnti-GPIIb/IIIa, GPIb/ IX and GP I a/II a antibodies were assayed by ELISA for patients with ITP. Total 442 patients in our hospital, who were retrospectively investigated from December 2010 to November 2012, were divided into newly diagnosed ITP, persistent and chronic ITP. The expression of specific anti- platelet glycoprotein antibody in each group was measured separately. The newly diagnosed ITP patients were treated with intravenous IgG (IVIG) and corticosteroids. The relationship between the expression of specific anti- platelet glycoprotein antibodies GPIIb/IIIa, GPIb/IX and GPIa/IIa and the complete response (CR) was studied.

RESULTSPositive rates of anti- platelet glycoprotein antibodies were 59.09%, 26.97% and 37.35% respectively in newly diagnosed ITP, persistent and chronic ITP, the difference was statistical significant (P<0.05). In newly diagnosed ITP, positive rate of antibody against GPIIb/IIIa was 38.64%, double positive rate of antibodies against both GP II b/III a and GP I a/II a was 15.91%, there was statistical significance (P<0.05) compared with that of persistent and chronic ITP. The complete response (CR) rate in newly diagnosed ITP patients with positive antibody against GP II b/III a was 80.39% after treatment with IVIG and corticosteroids. There was statistical significance compared with that in patients having no antibodies (P<0.05).

CONCLUSIONThe expression of antibodies against GP II b/III a and double positive for both GP II b/III a and GP I a/II a autoantibodies increased in newly diagnosed ITP patients. Patients with anti-GP II b/III a autoantibody had good response to medication with IVIG and corticosteroids.

Adolescent ; Adult ; Aged ; Autoantibodies ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Platelet Glycoprotein GPIb-IX Complex ; immunology ; Platelet Membrane Glycoproteins ; immunology ; Retrospective Studies ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Treatment Outcome ; Young Adult

Objective: To evaluate the efficacy and safety of a domestic human plasma derived coagulation Factor Ⅸ concentrate (pd-FⅨ) in patients with hemophilia B. Methods: The study was a multicenter, open-label and single-arm study. The efficacy of pd-F Ⅸ was evaluated by objective performance criteria. The doses of pd-FⅨ were calculated according to the bleeding symptom and disease severity. The infusion efficiency of pd-FⅨ and improvement of bleeding symptoms were measured at 30 minutes and (24±4) h after the first infusion, respectively. Adverse events were recorded. Viral infection and FⅨ inhibitor were detected 90 d after the first infusion. Results: All 36 subjects with hemophilia B were enrolled in the study. The median age of these patients was 31 years old and the median injection doses were 4 (1-17) times. The hemostatic effect of 27/36 (75.00%) and 9/36 (25.00%) acute bleeding events were rated as "excellent" and "better" , respectively. The recovery rate was 111.92% (65.55%-194.28%) at 30 minutes after infusion of FⅨ. There was no adverse event related to FⅨ. No reactivation of HBV, HCV or HIV and FⅨ inhibitor was detected at 90-104 d after the first FⅨ infusion. Conclusion: This domestically made human plasma derived FⅨ concentrate is safe and effective in the treatment of acute bleeding in patients with hemophilia B. Clinical trial registration: China food and Durg Administration, 2016L08027.
Adult ; China ; Factor IX ; Hemophilia A ; Hemophilia B/therapy* ; Hemorrhage ; Humans ; Plasma