Objective To pursue a more efficient and effective treatment for cleft lip and palate deformities. Methods Patients with unilateral complete cleft lip and palate at their age of 9 years after were chosen for simultaneous cleft alveolar repair and nasal deformity correction. Muco-periosteal pocket and iliacgranular bone was prepared, and bone grafting was performed conventionally. At the same time of iliac cancellous bone harvesting, a cortical plate was taken and sculpted into a strut of 18 mm in length, 6 mm in width, and 1.5 mm in thickness. A flying bird incision was made at the alar ram and across columella in a V-shape. Then the alar cartilage was detached from the overlying skin, a socket was made at the site of anterior nasal spine. The strut was inserted into the socket between the two medial crura of the alar cartilage. The medial crura was lift 3 mm above the superior edge of the strut, and mattress suture technique was used to secure the bilateral medial crura to the strut graft. Results 24 patients were treated by this technique. All the patients healed uneventfully. Depressed alar base, tilted columella and lower nasal tip were corrected satisfactorily. Conclusion There is no interference in simultaneous cleft alveolar bone grafting and rhinoplasty. Septum strut can provide favorable support for tilted nasal structure and satisfactorily correct nasal deformities. Simultaneous with alveolar grafting, it is much easier in harvesting, and the time of anesthesia and operation is also decreased.[

Objective: To explore the in vitro inhibition effect of Zuojin pills on 6 cytochrome P450 isoenzymes ( CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) in human liver microsomes. Methods:The water extract of Zuojin pills, Cop-tidis Rhizoma and Euodiae Fructus was respectively incubated with human liver microsomes in the presence of seven probe substrates of CYP450 isoenzymes, and seven metabolites of cytochrome P450 probe substrate ( paracetamol/CYP1A2, 6α-hydroxypaclitaxel/CYP2C8, 4-hydroxydiclofenac/CYP2C9, 4-hydroxymephenytoin/CYP2C19, dextrorphan/CYP2D6, 6β-hydroxytestosterone/CYP3A4 and 1-hydroxymidazolam/CYP3A4) were simultaneously measured by LC-MS/MS, and the inhibitory effects were evaluated with IC50 value. Results:The IC50 value of Zuojin pills on CYP2D6, CYP1A2 and CYP3A4_T was 11. 6, 77. 4 and 97. 0 μg·ml-1 , respec-tively. The other IC50 values were from 334 to 690μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Coptidis Rhizoma on CYP2D6, CYP1A2 and CYP3A4_T was 5. 8, 36. 8 and 59. 2 μg·ml-1 , respectively. The other IC50 values were from 163 to 476 μg·ml-1 on CYP2C8, CYP2C9, CYP2C19 and CYP3A4_M isoenzymes. The IC50 value of Euodiae Fructus on CYPs was over 107 μg·ml-1 . Conclusion:Zuojin pills shows notable inhibitory effect on CYP2D6, and weak inhibitory effects on CYP1A2 and CYP3A4_T. Coptidis Rhizoma has similar effects on CYPs and may be the main herbal medicine in the formu-la. Therefore, much attention should be paid to the combination of Zuojin pills and the drugs metabolized by human CYP2D6 in clin-ics.

ObjectiveTo observe the influence of calcium - sensing receptor (CaSR) on Fas/Fas ligand (Fas L) pathway during anoxia/reoxygenation (A/R)- induced cardiomyocytes apoptosis in neonatal rat. MethodsSingle cells were dissociated from minced hearts of 2 - day - old Wistar rats with a 0. 25％ solution of crude trypsin and then cultured as monolayers at a density of 5 x 104cells/cm2 in DMEM medium equilibrated with humidified air containing 5％ CO2 at 37C. Three days after the cells were seeded, the cultured cardiomyocytes were randomly divided into three groups. ( 1 ) control group: cardiomyocytes were continuously cultured for 26 hours in DMEM medium.(2) A/R group: cardiomyocytes underwent anoxia for 2 hours and reoxygenation for 24 hours.(3) GdCl3 group: 300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation. Apoptosis of cardiomyocytes was assessed by flow cytometer and morphological alterations were observed with transmission electron microscope.The expression of CaSR, Fas and Fas L were analyzed by Western blot.Results The result of flow cytometer showed that cardiomyocytes apoptosis was 12. 18％ ± 1.54％ in A/R group,and was higher than that in the control group (P ＜0. 01 ). At the same time, mitochondrial cristae dissolution and disappearance could be detected. Compared with A/R group, GdCl3, a specific activator of CaSR, further enhanced cardiomyocytes apoptosis to 20. 25％ + 2. 87％ ( P ＜ 0. 01 ), along with an increment in CaSR, Fas and Fas L expressions ( P ＜ 0. 05 ). ConclusionCaSR is closely involved in cardiomyocytes apoptosia during anoxia/reoxygenation. CaSR could induce apoptosis of neonatal rat cardiomyocytes through Fas/Fas L receptor pathway.

Objective:To examine the osmolality of domestic recombinant human interferon α2b injection to provide evidence for the improvement of the national quality standard. Methods:Totally 66 batches of recombinant human interferonα2b injection produced by 9 manufacturers were withdrawn, and the osmolality was determined according to the appendix of Chinese Pharmacopoeia Ⅲ(2010 edition). The results were analyzed with statistical methods. Results:The pass rate of osmolality was 98. 5%. The osmolality of more than 90% of the batches was between 85% and 115% of the intermediate value set by the manufacturers. Conclusion:Comprehensive understanding of the quality control of osmolality of domestic recombinant human interferon α2b injection is obtained, which provides data support for the improvement of quality standard of osmolality.

To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.

Objective To explore the relationship between G-protein β3 subunit (GNB3) gene C825T polymorphism and antipsychotic agent-induced obesity.Methods 126 schizophrenic inpatients with long-term antipsychotics treatment were collected.According to body mass index ( BMI),patients were divided into obesity group ( n =62) and non-obesity group ( n =64).The GNB3 gene C825T polymorphism was detected by polymerase chain reaction and DNA sequencing technique.Levels of fasting blood glucose,2-hour postprandial blood glucose,blood lipids and blood uric acid of all patients were routinely measured.Results (1)The GNB3 gene C825T polymorphism were found in obesity group and non-obesity group respectively,and the distribution of genotypes in two groups were both consistent with Hardy-Weinberg equilibrium.(2)There was no significant difference in genotype frequencies between obesity group ( CC 17.75％,CT 58.06％,TT 24.19％ ) and non-obesity group( CC 18.75％,CT 62.50％,T T 18.75％ )( x2 =0.59,P ＞ 0.05 ).There was also no significant difference in allele frequencies between obesity group ( C 46.77％,T 53.23 ％ ) and non-obesity group ( C 50％,T 50％ ) ( x2 =0.26,P ＞ 0.05 ).(3)No significant differences were observed in BMI,fasting blood glucose,2-hour postprandial blood glucose,blood lipids and blood uric acid among different genotype groups (all P ＞ 0.05 ).Also no significant differences were observed in BMI,fasting blood glucose,2-hour postprandial blood glucose,blood lipids and blood uric acid between Tallele carrier (TT and CT genotypes) and T-allele non-carrier( CC genotype) ( all P ＞ 0.05 ).Conclusion The GNB3 gene C825T polymorphism may not be a genetic risk factor for antipsychotic agent-induced obesity.

Accurate determination of biological activity is essential in quality control of recombinant human brain natriuretic peptide (rhBNP). In previous study, we successfully developed a genetically modified cell line 293GCAC3-based ELISA assay for rhBNP. But ELISA procedure is still tedious, so this study was aimed to develop a rapid and simple bioassay for rhBNP using GloSensor technology, which provides a platform of flexible luciferase-based biosensors for real-time detection of signaling events in live cells, including cGMP production. A reporter cell line 293GCAGlo-G1 was constructed by transfecting pGloSensor?40 F plasmid into 293GCAC3. The reporter assay based on 293GCAGlo-G1 showed high precision with intra-assay CV being 8.3% and inter-assay CV being 14.1%; high accuracy with 80%, 100% and 120% recovery rate being 99.2%, 102.4% and 99.0% respectively; and great linearity with R2of linear fitting equation being 0.99. Besides, no significant difference was found in test results of reporter assay and 293GCAC3-based ELISA assay (paired t test, p＝0.630). All these results suggested that the reporter assay was a viable assay for biological determination of rhBNP.

OBJECTIVE: To investigate the expression of Bax and PHF20 in laryngeal squamous cell carcinoma (LSCC)and to discuss their relevance and the roles in carcinogenesis and development in LSCC. METHOD: The expressions of Bax and PHF20 in the LSCC tissues and normal mucosa tissues adjacent to carcinoma were detected by SP immunohistochemistry assay. The relationship between the expressions of Bax and PHF20 and the clinicopathological characteristics including clinical stage, pathological type, histological grade and lymph node metastasis in LSCC were analyzed according to the clinical data. RESULT: (1) The expressions of Bax and PHF20 were both significantly lower in the LSCC tissue than that in the normal laryngeal tissue (P < 0.01). (2) In clinical stage grouping, there were no statistical differences of the quantity and positive rate of Bax and PHF20 expressions among supraglottic, glottic and subglottic LSCC (P > 0.05). In histological differentiation grouping, the quantity and positive rate of Bax and PHF20 expressions decreased significantly in poorly differentiated LSCC compared with the well and moderately differentiated LSCCs (P < 0.01, P < 0.05, respectively). In T stage grouping, the quantity and positive rate of Bax and PHF20 expressions were both significantly higher in T1 + T2 compared with T3 +T4 (both P < 0.01). In addition, the quantity and positive rate of Bax and PHF20 expressions were both significantly higher in LSCC with lymph node metastasis compared to that without lymph node metastasis (both P < 0.01). CONCLUSION The lack of Bax and PHF20 might contribute to the carcinogenesis and development in LSCC. The positive expression of Bax and PHF20 maybe relative to T term degree, differentiation degree and lymphamatic metastasis of LSCC.
Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Head and Neck Neoplasms ; metabolism ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; Lymphatic Metastasis ; Prognosis ; Squamous Cell Carcinoma of Head and Neck ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE: To determine the sensitivity of cetuximab induced apoptosis in laryngeal squamous carcinoma cells Hep-2, and to evaluate the synergistic killing effects and regulation mechanism of cetuximab alone or cetuximab in combination with chemotherapeutic agents (cisplatin) or radiation means on Hep-2 cells. METHOD: To investigate the cytotoxicities of cetuximab, cisplatin and radiation, cell counting kit-8 (CCK-8) assay was used for the detection of cell growth inhibition ratio, and fluorescence activated cell sorter FACS for the apoptotic rate and cell cycle distribution. RESULT: Cetuximab had inhibitive effect on Hep-2 cells within a certain range of concentration in a time- and dose-dependence manner. The inhibition concentration 50% (IC50) of cetuximab on Hep-2 cells for 24 h was 1 036.84 microg/ml. For application of cisplatin and radiation, the apoptotic rate of Hep-2 cell was higher by combining with cetuximab than their single or combined administration. Moreover, the cell cycle arrested at G0/G1 phase. CONCLUSION Laryngeal cancer Hep-2 cells was sensitive to the cetuximab induced apoptosis. Cetuximab combined with cisplatin and/or radiation can increase the antiproliferative effects on Hep-2 cells. These findings suggest the synergistic combination of cetuximab and cytotoxic agents was sequence depended.
Antibodies, Monoclonal, Humanized ; pharmacology ; Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Cetuximab ; Cisplatin ; pharmacology ; Combined Modality Therapy ; Humans

The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.