Background Perineural invasion is an important biological character for adenoid cystic carcinoma (ACC) of lacrimal gland,which is different from those of other lacrimal gland tumors.As the important part of neurotrophic factors,glial-derived neurotrophic factor (GDNF) plays an important role in perineural invasion for ACC of salivary gland.GDNF regulation in the ACC cell biology function needs to be further explored.Objective This study was to investigate the effect of GDNF on proliferation and migration of ACC cells,and to explore the mechanism of neural invasion in ACC of lacrimal gland.Methods ACC-2 cell line was cultured and passaged in RPMI 1640 medium with 10％ fetal bovine serum,100 U/ml penicillin and 0.1 g/L streptomycin.Single-cell suspension was prepared with the density of 2×104/ml using logarithmic phase of cells and then incubated to 96-well plate.GDNF with the final concentration of 20,60,80,100 and 120 μg/L was added into the medium respectively in the experimental groups,and the cells were cultured in the medium without GDNF as the control group.The expression of GDNF in ACC-2 cells was detected by immunohistochemistry.MTT assay was employed to assay the absorbance value at the wavelength of 570 nm (A570) for the evaluation of proliferation of ACC-2 cells after cultured by different concentrations of GDNF for different time points.Meanwhile,transwell chamber was used to examine the cell migrated number.Results Immunochemistry assay exhibited that ACC-2 cells showed the positive response for GDNF with the brown staining in the cytoplasm.In 48 hours after culture,the A570 value was elevated with the increase of GDNF concentration,showing a significant difference among various groups (F =3.336,P =0.026),and the A570 value in various concentrations of GDNF groups was higher than that of 0 μg/L GDNF group (all P＜0.05).After action of 80 μg/L GDNF,the A570 value of the cells was gradually increased with the prolong of culture time (Ftime =39.979,P=0.000).In 30 minutes after GDNF cultured,the number of migrated cells increased with the increase of GDNF concentration (F=144.886,P=0.000).ACC-2 cells were cultured by 100 μg/L GDNF for 24,30 and 40 hours,the number of migrated cells were more as the time lapse,and more migrated cells were seen in GDNF group at various time points (Ftime =46.747,P =0.000 ; Fgroup =63.786,P =0.000).Conclusions GDNF can stimulate the proliferation and migration of ACC-2 cells in a dose-and time-dependent manner.

Information Systems ; Medical Informatics ; Medical Records Systems, Computerized ; Pathology

Objective To investigate the correlation between the incidence and characteristics of anemia in chronic heart failure population through retrospective analysis of patients hospitalized at the Hunan Provincial People's Hospital.Methods A total of 259 patients who were diagnosed as chronic heart failure (CHF) and hospitalized at the Hunan Provincial People's Hospital from October to November in 2012 was collected,and divided into cardiac function Ⅰ ～ Ⅳ grade,then their clinical characteristics was analyzed.Results (1)Compared with CHF patients without anemia,the creatinine value was significantly higher,the percentage of patients with severe heart failure was significantly higher in the CHF patients with anemia (P＜ 0.05).(2) Logistic regression analysis demonstrated that age (95％ CI:1.123 ～ 3.580,P ＜ 0.05),renal insufficiency (95％ CI:1.320 ～ 4.845,P ＜ 0.05),and cardiac function (95 ％ CI:1.368 ～ 3.385,P ＜ 0.05) in patients with heart failure were important risk factors for anemia.Conclusions Anemia was common in CHF patients.The poorer cardiac function was and the lower hemoglobin was,the higher incidence of anemia was.The patients with anemia had the characteristics of worse renal function and poorer nutritional status compared patients without anemia.Age,renal insufficiency,and cardiac function were important risk factors for anemia.

In the reform of medical microbiology education on medical students of 8-year-education pro-gram, we compared the traditional teaching methods with Problem Based Learning (PBL) method. Through our practice, we have found that the combination of traditional lecture-based learning and PBL seems to better match the students’ way of learning. The lack of basic knowledge of the students hindered their learn-ing effect during the bilateral discussion in the PBL education. We also found that the application of PBL in medical microbiology education is an iterative process and should be promoted step by step. The theoretical level and the innovative ideas of the teachers play a crucial role in the dynamic process of education reform.

Objective To evaluate the impact of chemotherapy compliance on the therapeutic efficacy of induction chemotherapy plus concurrent chemoradiotherapy versus induction chemotherapy plus radiotherapy alone for patients with locally advanced nasopharyngeal carcinoma (NPC). Methods Based on intention to treat analysis (ITT) for 400 patients, 314 patients were analyzed by per protocol (PP) analysis. The patients were divided into induction chemotherapy plus concurrent chemoradiotherapy group (IC/CCRT, 127 patients) or induction chemotherapy plus radiotherapy group (IC/RT, 187 patients). The patients who completed 2 cycles of induction chemotherapy and at least 2 cycles of concurrent chemotherapy in the IC/CCRT group and the patients who completed 2 cycles of induction chemotherapy in the IC/RT group were analyzed. Radiotherapy was given by two-dimensional technique with γ-ray, X-ray and electron beams. The chemotherapy regimen was FUDR plus carboplatin for induction chemotherapy and carboplatin alone for concurrent chemotherapy. Results The follow-up rate was 96.2%. 295 patients were followed to at 3 years. Based on PP analysis, Grade 3/4 toxicity was found in 23.6% of the patients in IC/CCRT group and 13.4% in the IC/RT group (χ~2 =5,50,P=0.019). No grade 4 toxicity was found in the IC/RT group. The median follow-up time was 3.9 years, and no significant difference was found between the two groups in 3-year overall survival (78.1% : 84.6% ;χ~2 = 0. 61, P =0. 435), disease-free survival (74.3 % : 70.1% ;χ~2= 0. 12, P= 0.731), Iocoregional relapse-free survival (89.7% : 89.5% ; χ~2= 0. 10, P= 0.748), or distant metastasis-free survival (78.9%:76.5% ;χ~2=0.05,P=0.825). Conclusions With more severe toxicities, the IC/CCRT regimen does not improve the overall survival in locally advanced NPC patients compared with the IC/RT regimen.

Objective To compare the efficacy of concurrent chemoradiotherapy versus radiotherapy alone for T3-4 N0-1 M0 and T14 N2-3 M0 nasopharyngeal carcinoma (NPC) after induction chemotherapy.Methods From 2002 to 2005,400 patients with stage Ⅲ and Ⅳa NPC were randomly divided into 2 groups :induction chemotherapy followed by concurrent chemoradiotherapy group (IC/CCRT,201 patients),and induction chemotherapy followed by radiotherapy alone group (IC/RT, 199 patients).Subgroup analysis was conducted for 197 patients with stage T3-4N0-1M0 NPC and 203 with stage T1-4N2-3M0 NPC.Results The follow-up rate were 96.2%, with a median followg-up time of 3.9 years.For T3-4N0-1 M0 NPC patients in IC/CCRT group (104 patients) and IC/RT group (93 patients), the 3-year overall survival, disease-free survival, locoregional recurrence-free survival and distant metastasis-free survival rates were 84.0% and 85.9% (χ2=0.08,P =0.780) ,77.0% and 72.0% (χ2=0.44,P =0.510) ,89.5% and 92.3% (χ2=0.65 ,P = 0.420), and 84.9% and 77.0% (χ2= 1.59, P = 0.210), respectively; For T1-4 N2-3 M0 NPC patients in IC/CCRT group (97 patients) and IC/RT group (106 patients), the corresponding rates were 67.4% and 82.2% (χ2=3.48,P=0.060), 61.5% and 68.0% (χ2= 1.86,P=0.170), 86.2% and 87.0% (χ2=0.57 ,P =0.450) and 66.2% and 75.6% (χ2=2.07 ,P =0.150), respectively.Acute sideeffects were similar except more leucocytopenia in IC/CCRT group than IC/RT group.Conclusions Compared with IC/RT, IC/CCRT dose not improve the overall survival in patients with T3-4N0-1 M0 and T1-4 N2-3 M0 NPC.

OBJECTIVETo research the application and clinical effect of titanium elastic nails (TEN) for the treatment of closed clavicular fracture in elder children.

METHODSFrom October 2010 to December 2012,16 elderly children with clavicular fracture of elder children were treated with internal fixation, including 9 boys and 7 girls with an average age of 14.2 yeats old ranging from 9 to 17 years. The mean time between injury and surgery was 2 to 6 days. Thirteen patients had a transverse fractures; 3 patients had a oblique fractures, excluding the long-oblique patients. Neer function score after 3 months follow-up and the shoulder activity degree after 3 weeks followed were compared respectively between the injured side and healthy side. Fracture reduction and healing were followed up by X-rays to analyze internal fixation with TEN technique.

RESULTSAll patients were healed well after surgery. There were 2 cases with soft tissue irritation. No cases of infection, TEN broken, delayed healing or skin bursting were observed. All patients were followed up for 3 to 10 months,with an average of 7.2 months. There were no significant differences (t=2.12, P>0.05) between the healthy side (98.75±2.70) and injured side(96.69±6.22)of Neer score at 3 months during followed up. The shoulder activity at 3 weeks during followed up was different between both sides (P<0.05). The intramedullary nail was removed after X-rays at 12-18 weeks during followed up showed good bone healing,and the shoulder function was recovered well.

CONCLUSIONInternal fixation with TEN for the treatment of clavicular fracture has advantages of a limited invasion, beauty, safety and reliability. This technique provides an ideal fixation option for the treatment of clavicular fracture in elder children.

Adolescent ; Bone Nails ; Child ; Clavicle ; injuries ; surgery ; Elasticity ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Titanium

The study of mammary gland bioreactor is in the ascendant. In order to generate transgenic goats of well-controlled expression of exogenic genes, we constructed a human lactoferrin (hLF) gene targeting vector containing promoter, exon 1, intron1 and some of exon 2 (about 6.1 kb fragment) and exon 6 approximately 9 (about 3.3 kb fragment) of the goat beta-casein gene as well as hLF minigene, neo gene inserted into them and tk gene ligated to the 3' end of the construct. The 9.4 kb goat genomic sequences as homologous arms were initially amplified by PCR with local goat tissue DNA. The expression vector was named pBC-tk-neo-hlf. Then the recombinant plasmid pBC-tk-neo-hlf containing hLF minigene was transfected into mice mammary tumor cell line C127 by liposome, cell clones were selected with G418. After proliferating, the transfected cells were induced with insulin, luteotropic hormone and hydrocortisone. The result of Western-blotting analysis showed that the transfected cells can secrete hLF protein, and the recombinant protein expressed in cultured cell supernatant has the similar molecular weight as the native protein. The expression level detected by ELISA was 0.21 microg/mL. This result indicated that the targeting vector could efficiently direct the expression of hLF in mammary cells,and it confirmed the validity of the constructed vector. At the same time, C127 cell line proved to be useful for evaluating the regulation of a foreign gene expression in mammary gland specific expression vector.
Animals ; Caseins ; genetics ; Cell Line, Tumor ; Goats ; Humans ; Lactoferrin ; genetics ; Mammary Glands, Animal ; cytology ; metabolism ; Mice ; Molecular Weight ; Transfection

Objective To study the therapeutic effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on acute liver failure ( ALF) induced by D-galactosamine (D-gal) and lipopolysaccharide (LPS) in rats. Methods The ALF model was obtained through intraperitoneal injection of D-gal(300 mg/kg)and LPS (20μg/kg)in Wister rats. The hUCMSCs were transplanted after intoxication. All rats were divided into four groups, and each group received either hUCMSCs or 0.9% NaCl solution through intraperitoneal or tail-intravenous injection. To evaluate the liver function of each group, the levels of alanine aminotransferase (ALT), total bilirubin (TBil) and serum albumin (Alb) were measured on the day of hUCMSCs transplantation and the following 1, 2, 3, 5 and 7 days. All rats were then sacrificed to examine the liver histology at day 7. Analyses were done by using Fisher's exact test, unpaired t test and Mann-Whitney U test. Results There were no significant differences of survival rates among four groups (Fisher's exact test, both P = 1. 00). The levels of ALT, TBil and Alb in group receiving hUCMSCs intraperitoneally were (804. 9 ± 88. 0) U/L,(17. 4±2. 7) μmol/L and (20. 9±0. 8) g/L, respectively after 2 days of injection, whereas in the corresponding control group, those were (1294. 3± 171. 4) U/L, (32. 3±5. 5) μmol/L and (16. 1±0. 9) g/L, respectively, which indicated that hUCMSCs transplantation significantly improved the liver function (t = 2. 640, P =0.020;t=2.529, P = 0. 025;t= - 3. 833, P = 0. 002). Both of hUCMSCs-transplanted groups showed no significant differences. Liver histological data showed that transplantation of hUSMSCs through either intraperitoneal or tail-intravenous injection alleviated liver damage (U=4. 500, P = 0. 005;U=4. 500, P = 0. 008) and the mitotic index also increased in hUCMSCs-transplanted groups (U=4. 000, P = 0. 005; U=5. 500, P = 0. 013). Conclusions The levels of ALT, TBil and Alb can rapidly normalize in ALF rats after injected with hUCMSCs either intraperitoneally or tail-intravenously. hUCMSCs application raises the mitotic index, enhances hepatocellular regeneration and improves histological status.

OBJECTIVETo investigate the effects of experimental occlusal hypofunction on the architecture of rat's mandibular bone.

METHODSForty Sprague-Dawley (SD) male rats were randomly divided into experimental occlusal hypofunction group and control group, with 20 rats in each group. Experimental occlusal hypofunction group rats were fitted with a metal cap to the maxillary and mandibular incisor, to eliminate the occlusal forces on rat molars. In control group, a metal ring was fitted to the maxillary and mandibular incisor to maintain molar occlusal contact. Then the rats were killed under deep anaesthesia at 0 week and at the end of 2, 4, 6, and 8 weeks, and the mandibles were immediately removed for measurement of the three-dimensional architecture of cancellous bone in the alveolar process with micro-CT.

RESULTSAt the end of week 4, the bone volume/total volume (BV/TV) in the hypofunction group decreased compared with the control group [control group:(0.65 ± 0.05)%, experimental group: (0.60 ± 0.05)%, P < 0.05] and lasted until the end of the experiment [control group: (0.72 ± 0.06)%, experimental group: (0.51 ± 0.07)%, P < 0.01]. The similar result was also observed in trabecula thickness (Tb. Th) in experimental animals [control group: (168 ± 15), (170 ± 25), (180 ± 18) µm, experimental group: (152 ± 20), (145 ± 19), (142 ± 15) µm, P < 0.05]. The trabecula spacing (Tb. Sp) in the hypofunction group increased from the end of week 4 to the end of week 8 compared with the matched control groups [control group: (264 ± 21), (284 ± 17), (282 ± 26) µm, experimental group: (306 ± 30), (316 ± 18), (332 ± 18) µm, P < 0.05]. The trabecula number (Tb.N) of experimental animals decreased from the end of week 6[control group:(3.59 ± 0.22) mm(-1), experimental group: (3.03 ± 0.31) mm(-1), P < 0.05] to the end of week 8[control group: (3.66 ± 0.24) mm(-1), experimental group: (2.85 ± 0.18) mm(-1), P < 0.01].

CONCLUSIONSExperimental occlusal hypofunction can result in the change of mandibular alveolar process three-dimensional architecture.

Animals ; Bite Force ; Bone Density ; Dental Occlusion ; Imaging, Three-Dimensional ; Male ; Mandible ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; X-Ray Microtomography