Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

BACKGROUNDCervical arthroplasty is indicated to preserve cervical motion and prevent accelerated adjacent segment degeneration. Whether accelerated adjacent segment degeneration is prevented in the long term is unclear. This trial compared adjacent segment degeneration in Bryan disc arthroplasty with that in anterior cervical decompression and fusion five years after the surgery.

METHODSWe studied patients with single level degenerative cervical disc disease. The extent of adjacent segment degeneration was estimated from lateral X-rays.

RESULTSTwenty-six patients underwent single level Bryan disc arthroplasty and twenty-four patients underwent single level anterior cervical decompression and fusion. All patients were followed up for an average of sixty months. In the Bryan arthroplasty group, nine (17.6%) segments developed adjacent segment degeneration, which was significantly lower than that (60.4%) in the anterior cervical decompression and fusion group. Eleven segments in the Bryan arthroplasty group developed heterotopic ossification according to McAfee's classification and two segments had range of motion less than 2°. In the heterotopic ossification group, four (19.5%) segments developed adjacent segment degeneration, similar to the number in the non-heterotopic ossification group (16.7%). Adjacent segment degeneration rate was 50% in grade IV group but 11.8% in grade II to III.

CONCLUSIONSAdjacent segment degeneration was accelerated after anterior cervical decompression and fusion. However, Bryan disc arthroplasty avoided accelerated adjacent segment degeneration by preserving motion. Patients with grade IV heterotopic ossification lost motion, and the rate of adjacent segment degeneration was higher than that in patients without heterotopic ossification.

Adult ; Arthroplasty ; adverse effects ; Case-Control Studies ; Cervical Vertebrae ; surgery ; Female ; Humans ; Intervertebral Disc ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; adverse effects ; Young Adult

OBJECTIVETo analyze the related factors of the infection after operation of open tibiofibula fractures.

METHODSThe clinical data of 141 patients with open tibiofibula fractures underwent surgical treatment from June 2009 to December 2012 were retrospectively analyzed. All the patients were male and aged from 18 to 61 years old with an average of 39.2 years; all the fractures were unilateral. According to Gustilo typing of fracure, 5 cases were type I, 44 cases were type II, 27 cases were type III a, 56 cases were type III b and 9 cases were type III c. These clinical data included patients' age, gender, body mass index (BMI), underlying diseases, time of operation, fracture site, fracture type, fixation method, postoperative drainage, debridement condition and so on. The postoperative infection conditions were recorded, and the correlation between above factors and infections were analyzed. Finally, the significant related variables were introduced into a Logistic regression model to evaluated their risk.

RESULTSAmong the 141 patients, 22 cases developed with infection (15.6%). The significant relative factors with infection contained fixation method, debridement condition, underlying diseases and postoperative drainage (P < 0.05). The correlativities were stepped up in order, their odds ratios value was 2.451, 3.164, 3.414, 5.117, respectively.

CONCLUSIONActive treatment for underlying diseases before operation, thorough debridement for open wound, suitable fixation method for fracture and thorough draining are effective measures in preventing postoperative infection.

Adolescent ; Adult ; Debridement ; Fibula ; injuries ; Fractures, Open ; surgery ; Humans ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Surgical Wound Infection ; etiology ; Tibial Fractures ; surgery

P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.

Objective To understand the effectiveness of China Infectious Disease Automated-alert and Response System(CIDARS)for outbreak detection at the regional level.Methods Two counties in Hunan province(Yuelu and Shuangfeng county)and two counties in Yunnan province(Xishan and Gejiu county)were chosen as the study areas. Data from CIDARS were analyzed on the following items: reported cases, warning signals, the time interval of signal response feedback, way of signal verification, outcome of signal verification and field investigation, from July 1,2008 to June 30,2010. Results In total, 12 346 cases from 28 kinds of diseases were reported,and 2096 signals of 19 diseases were generated by the system, with an average of 4.94 signals per county per week. The median of time interval on signal verification feedback was 0.70 hours(P25-P75:0.06-1.29 h)and the main way of signal preliminary verification was through the review of surveillance data(account for 63.07%). Among all the signals, 34 of them(1.62%)were considered to be related to suspected events via the preliminary verification at the local level. Big differences were found to have existed on the proportion of signals related to the suspected events of the total signals among the four counties, with Shuangfeng county as 4.71%, Yuelu county as 1.88%, Gejiu county as 0.95% and Xishan county as 0.58%. After an indepth study on the fields of suspected events, 12outbreaks were finally confirmed, including 5 on rubella, 4 on mumps, 2 on influenza and 1 on typhoid fever. Conclusion CIDARS could be used to assist the local public health institutions on early detection of possible outbreaks at the early stage. However, the effectiveness was different depending on the regions and diseases.

OBJECTIVETo establish a protocol of rapid clonal propagation of Dioscorea zingiberensis using mature leaves as explants.

METHODOut the optimal hormone combinations for callus induction, adventitious bud initiation, adventitious bud multiplication and rooting were found out by using MS medium with the macroelements at half strength as basal medium and studying the effects of BA, NAA and IBA on the processes.

RESULT AND CONCLUSIONMore than 80% of leaf explants formed callus when they were cultured on the media containing 1.0 - 5.0 mg x L(-1) BA and 1.0 - 2.0 mg x L(-1) NAA for 60 days; more than 60% of calli initiated adventitious buds within 50 days after they were transferred to the media containing 2.0 - 5.0 mg x L(-1) BA and 0.2 - 0.5 mg L(-1) NAA; all the adventitious buds rooted well after they were planted on the medium with 2.0 mg x L(-1) IBA for 20 days; the regenerated plantlets grew vigorously after they were transplanted and the survival rate was up to 80%.

Culture Media ; Dioscorea ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; growth & development ; physiology ; Plants, Medicinal ; growth & development ; Regeneration ; Tissue Culture Techniques ; methods

OBJECTIVETo investigate the characteristic of cervical lymph node metastasis of hypopharyngeal carcinoma and its influence to the prognosis.

METHODSOne hundred and eight hypopharyngeal carcinoma patients who accepted treatments in the 1st Affiliated Hospital of China Medical University from 1985 to 2000 were reviewed retrospectively. All of them accepted surgical treatment without pre-operative chemotherapy or radiotherapy. Stage was made according to the standard of International Union Against Cancer (UICC) in 1992. Specimens of the patients were carefully examined to confirm the primary site of the tumor and the distribution of cervical lymph node metastasis. Pathological differentiations of the tumor were classified into high, middle and low category. Kaplan-Meier method was used to estimate the 3rd, 5th years survival.

RESULTSThe rates of lymph node metastasis of was 45.8% for patients with TI and T2 disease, 79.8% for those with T3 and T4, and 75.0% (81/108)for the whole patients(P < 0.05). Patients with pyriform sinus cancer occupied 92.6% (100/108) of all the cases. Cervical lymph node metastasis rate of pyriform sinus cancer and posterior pharyngeal wall cancer were 74. 0% and 87. 5% respectively (P > 0.05). Cervical lymph node metastasis rate of patients with the high, middle and low differentiation tumor were 72. 2% , 67.6% and 85.7% respectively (P > 0.05). The 3rd and 5th years survival rates of all patients were 67.53% and 29.87% respectively. The occurrence of cervical lymph node metastasis was 76.5% in the level II and III, and 8.6% in the level V and VI. CONCLUSIONS Cervical lymph node metastasis rate of hypopharyngeal carcinoma is high. Cervical lymph node metastasis was one of the most significant prognostic factors of hypopharyngeal carcinoma. With the increase of the cervical node metastasis, the 3rd and 5th years survival of the patients declined gradually.

Adult ; Aged ; Female ; Humans ; Hypopharyngeal Neoplasms ; diagnosis ; pathology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck ; pathology ; Prognosis ; Survival Rate

The present study investigated the relationship between DNA-dependent protein kinase (DNA-PK) and radiosensitivity of nasopharyngeal carcinoma (NPC) cell lines. The dose-survival relationship for NPC cell lines, CNE1 and CNE2, was analyzed using clonogenic formation assay, the activity of DNA-PK of the two cell lines was measured using the Signa TECT DNA-PK assay kit, and the localization and expression of Kus (a heterodimer) and DNA-PKcs protein in CNE1 and CNE2 before irradiation and 15 min, 1 h, 6 h, 12 h, 24 h after 4 Gy irradiation were analyzed by immunofluorescence, laser scanning confocal microscope (LSCM) and Western blot. The results showed that the surviving fraction of CNE1 was higher than that of CNE2 at each dose. The DNA-PK activity of CNE1 was also significantly higher than that of CNE2 before and after irradiation (P<0.05), while the expression of total Ku70/Ku80 in CNE1 and CNE2 had no significant difference. Increasing translocation of Ku70 and Ku80 from the cytoplasm to the nuclei in the two cell lines was observed with increase of irradiation time as detected by Western blot, and the immunofluorescence of the DNA-PK complex subunits showed greater nuclear translocation in CNE1 than CNE2 after irradiation. The results suggest that the relatively higher radio-resistance of CNE1 correlates with the higher activity of DNA-PK as compared to that of more radiosensitive CNE2 (or lower radio-resistance) before and after irradiation. Thus, DNA-PK activity may be a useful predictor of radiosensitivity of NPC.
Carcinoma ; Cell Line, Tumor ; enzymology ; radiation effects ; DNA-Activated Protein Kinase ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; enzymology ; Radiation Tolerance

OBJECTIVETo explore the clinical feature, imaging and their diagnostic value for Joubert syndrome (JS).

METHODThe clinical data, imaging feature, and 31 references from China Biomedical literature database (CBMdise) were reviewed and analyzed.

RESULTThe age of onset of 32 patients including male 20 and female 12 ranged from 3 days to 6 years (mean 2.2 years). All the 32 patients with Joubert syndrome showed "slow growth" and "reduced muscle tension", 26 cases (81.3%) showed "gasp for breath", 26 cases (81.3%) showed "unusual motion of eyeball", 2 cases (6.3%) showed additional fingers (toes), 6 cases (18.8%) showed stretching tongue with agape. The typical imaging features of Joubert syndrome included "molar tooth sign", "midline cleavage" between cerebellar hemispheres and "bat-wing" like fourth ventricle, all the 32 patients with Joubert syndrome showed "midline cleavage", "molar tooth sign" was present in 29 cases (90.1%), and "bat-wing" like fourth ventricle in 30 cases (93.8%).

CONCLUSIONJoubert syndrome is a rare congenital brain malformation. The typical clinical manifestations included "gasp for breath", "reduced tension of muscle", "slow growth" and "unusual motion of eyeball", and at the same time the patients had the following typical imaging features of brain: "molar tooth sign", "midline cleavage" and "bat-wing" like fourth ventricle.

Abnormalities, Multiple ; Cerebellar Diseases ; diagnosis ; physiopathology ; Cerebellum ; abnormalities ; Child ; Eye Abnormalities ; diagnosis ; physiopathology ; Female ; Humans ; Kidney Diseases, Cystic ; diagnosis ; physiopathology ; Male ; Retina ; abnormalities ; physiopathology

AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P ＜ 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P ＜ 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.