OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

Objective To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.Methods Male Wistar rats were randomly divided into sham operation group (n =8),sham operation + apocynin treatment group (n =8),UUO operation group (n =8) and UUO operation+apocynin treatment group (n =8).Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery.Rats were sacrificed at 7th day.ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT),and the level of 8-iso-prostaglandin F2alpha (8-isoPGF2α) in renal tissue.Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4,α-smooth muscle actin(α-SMA),collagen Ⅰ (COL-Ⅰ) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).Results UUO rats with vehicle displayed increased oxidative stress,as measured by renal tissue 8-iso-PGF2α,accompanied with increased renal expression of NADPH oxidases (NOX2,1.5-fold and NOX4,1.7-fold,respectively),compared with sham-operated rats (P ＜0.05).Furthermore,vehicle treated UUO rats showed increased renal COL-Ⅰ and α-SMA levels,compared with sham-operated rats (P ＜ 0.05).ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P ＜ 0.05).Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7％) and NOX4 (-31.0％) in UUO rats,respectively,compared with vehicle treated rats (P ＜ 0.05).And significant decrease of COL-Ⅰ (-26.4％) and α-SMA expression (-80.0％) were also observed (P ＜ 0.05).The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P ＜ 0.01).Despite the pronounced dysregulation of pro-oxidative NOXs family,no compensatory increase of antioxidative enzyme activities occurred.Conclusion The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation,and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.

Objective To evaluate the predictability of ablated depth in photorefractive procedures by Zeiss MEL80 and VISX Star S4 excimer laser.Design Prospective,comparative case series.Participants 168 eyes of 84 people with myopia and cylinder.Main Outcome Measures Corneal thickness and ablated depth.Methods The thickness of cornea by Zeiss MEL80 and VISX Star S4 were measured by anterior segment OCT.The actuality ablated depth and calculated depth were compared.Results There were no significant differences between actuality ablated depth and calculated depth in Zeiss MEL80.And in VISX Star S4,the actuality ablated depth was more than calculated depth.Linear regression of changes in ablated depth on myopia and cylinder yielded the following formulaus:△ablated depth(VISX Star S4)= 2.324 *|myopia|+ 5.270 *|cylinder|-6.772.Conclusions Different excimer laser has different changes between actuality ablated depth and calculated depth.To guarantee the safety of LASIK,Ophthalmologists should know the characteristics of the excimer laser.

ObjectiveTo evaluate the effect of selective posterior rhizotomy(SPR),local corrective surgery and the training of rehabilitation on the relief of limb spasm secondary to cerebral palsy.MethodsThe "open window" of bilateral vertebral lamina resection, the the posterior roots of L2-S1 were separated from anterior roots and splited into rootlets. The rootlets sensitive to lower threshold of electrical stimulation were sectioned. The ratios of cut root,acorrding to muscle tension, myodynamia, body weight, muscle function,were estimated and all of them <30%. The local plastic operation was performed for nonimprovement of lower limb spasm and the training of rehabilitation began at postoperation. ResultsThe SPR have been performed on 28 cases. The rate of spasticity relief was 90% and the rate of functional improvement was 80% with follow-up. ConclusionsIt is a successful way to treat spastic cerebral palsy with SPR and procedural method.

BACKGROUND:Acellular dermal matrix possesses good flexibility and simple trimming.The intracutaneous or subcutaneous injection of acellular dermal matrix powder has fibroblast migration and collagen deposition.It has been widely used in plastic and reconstructive surgery.OBJECTIVE:To explore the feasibility of reconstructed acellular dermal matrix as a scaffold for chondrocyte implantation.DESIGN,TIME AND SETTING:Comparative observation.The study was performed at the Peking University Medical Department and Beijing Jishuitan Hospital between August 2003 and February 2007.MATERIALS:Neonatal calf dermis was provided by Beijing Yuanheng Shengma Biology Technology Research Institute.A total of 24 healthy adult SD rats,weighing 250 g,regardless of gender,and 36 New Zealand rabbits,aged 3 months,were selected.METHODS:①Calf full-thickness back skin was incubated with cell free buffer or ABS/AES for decellularization,followed by surface modification using growth factors.②Three rectangle skin flaps at two sides of the spinal cord of rats were made,and implanted with acellular dermal matrix.The implants were harvested at 2,6,and 12 weeks postoperatively.③The rabbits were divided into experimental and control groups.The cartilage was obtained from the left articular facet to isolate chondrocytes.The chondrocytes were seeded on the acellular dermal matrix.The cartilage defect was made on the right hind limb of experimentalrabbits,and implanted with acellular dermal matrix containing autoiogous chondrocytes.Biogel wass dropped on the surface of carrier.In the control group,the cartilage defect was made on the right hind limb of rabbits and the wound was sutured.Two rabbits from control group and 5 from experimental group were selected respectively at 4,12 and 24 weeks postoperatively.MAIN OUTCOME MEASURES:Cross-linking effect comparison;repair effect of rabbit bone defects.RESULTS:①The acellular dermal matrix cross-linked by glutaraldehyde demonstrated an obvious inflammatory reaction with tissue bleeding and necrosis.Conversely,ADM treated with water-soluble cross-linking agent caused displayed good histocompatibility.②The cartilage defects were repaired completely;the attached cells survived and proliferated and the acellular dermal matrix was degraded after 24 weeks of surgery.CONCLUSION:The acellular dermal matrix decellularized with cell free buffer,digested with digestive buffer,Cross-linked by water-soluble cross-linking agent,and further decorated with growth factor exhibited good histocompatibility,and was suitable forcell attachment and growth.The acellular dermal matrix scaffold almost degrades in the rabbits,with no rejection,and the bone defects were repaired after 24 weeks.

Objective To summarize the methods of diagnosing and treating solid-pseudopapillary neoplasm of pancreas SPN so as to provide reference for its early clinical diagnosis.Methods We collected the clinical data of 62 patients with pathologically confirmed diagnosis of SPN treated in the First Affiliated Hospital of Xi'an Jiaotong University between Feb.2004 and Sep.2014.Then we retroactively analyzed and summarized the clinical characteristics,diagnosis and treatment of the disease.Results Among the 62 patients,55 were female and 7 were male;the mean age was (31.58±12.67) years old.The clinical features showed no specificity and the tumor was mostly detected upon physical check-up.The tumor was seen to be located mostly in the body and tail of the pancreas,and the average maximum diameter was (7.81±3.54)cm.We did not find obvious abnormality in routine pre-operative blood test results or liver and kidney functions.The imageological examination indicated tumor occupation in the pancreas.All the patients underwent surgical resection with no complications or death and had a good recovery after operation.Pathological diagnosis after operation was SPN without metastasis in lymph modes.Until the last time we followed up all the patients,we found relapse in two patients (3.2％).Conclusion SPN is a tumor that tends to affect young and middle-aged females.The malignancy grade of SPN is low,and patients with this disease show no specificity in clinical manifestation.The imageological examination is of vital importance in diagnosing SPN and surgical resection is an effective way to treat SPN.

Objective To evaluate the feasibility of composite material of bone morphogenetic protein (BMP) and PLGA/TCP in repair of peri-porous-titanium bone defects in adult rabbits.Meth- otis The composite of PLGA/TCP scaffold with bovine BMP was made and implanted in distal bone de- fects peri-porous-titanium in femur of adult rabbits.The effect of BMP with PLGA/TCP on peri-prosthesis was evaluated by means of scanning electron microscope (SEM),energy dispersive X-ray analysis (EDX) and biomechanics.Results BMP with PLGA/TCP showed obvious peak of Ca and P of EDX in the pores of titanium in experiment group six weeks after operation and higher than that in control group (P＜0.05).Push-out test demonstrated that the bonding strength between the composite of HA coating/ porous titanium and bone was increased significantly with time (P＜0.05).In experiment group,at 6 and 12 weeks,peri-porous-titanium had higher shearing force compared with control group (P＜0.05). Conclusion BMP loaded with PLGA/TCP is a promising bone graft for bone defects in revision arthro- plasty,as indicates that bone induction of BMP plays an important role in biological stabilizaiton.

Abstract?AIM: To compare the changes in epithelial thickness profile following TransPRK and Epi-LASIK for myopia.? METHODS: In this prospective non -randomized controlled study, 76 right eyes of 76 myopic patients with the spherical equivalent refraction -1.25 to -6.00D were included under the informed consent. The eyes were divided into TransPRK group for 43 eyes and Epi-LASIK group for 33 eyes. Epithelial thickness was measured using spectral-domain optical coherence tomography at different corneal zones ( central, 2mm; paracentral, 2-5mm;and mid-peripheral, 5-6mm) preoperatively and at 1, 3, and 6mo postoperatively. The results were compared between the two groups.?RESULTS: The epithelium were thicker at 3 and 6mo after surgery compared to preoperative measurements in the two groups (all P<0.05).In TransPRK group, the epithelial thickness at 3 and 6mo demonstrated a negative meniscus-like lenticular pattern with lesser thickening centrally and progressively great thickening centrifugally (F3mo =-2.687,P=0.027;F6mo =-2.908,P=0.000).No statistically significant change was detected among the three zones in Epi-LASIK group (F=1.365, P=0.237). The epithelial thickness was thicker in the TransPRK group compared to the Epi-LASIK group mid-peripherally ( P<0.05) .? CONCLUSION: Significant epithelial thickening was observed after TransPRK and Epi-LASIK.It was showed a lenticular change with more thickening mid-peripherally after TransPRK than Epi -LASIK. Wound healing and inflammation may account for differences in the effect on epithelial thickness change by both surgeries.

Objective To analyze the autoimmune status of kidney in transgenic mice with high serum titer of anti-keratin autoantibody(AK auto Ab).Methods AK auto-Ab transgenie mice were set-up with conventional procedures,and the mice older than 20 weeks were selected for study.Urinary protein was quan- tified by Bradford method,and SDS-PAGE electrophoresis was used to analyze the type of proteinuria.Speci- mens of kidney were subjected to HE and PAS stain and immunofluorescence analysis and electrum microscopy observation were also performed.Results Compared with littcrmates,transgenie mice showed higher urinary proteins level(P

Sixteen compounds have been isolated from the EtOAc fraction of 95% ethanolic extract of Sophora dunnii through silica gel, Sephadex LH-20 and semi-prerarative HPLC column chromatographies. Their structures were identified on the basis of NMR and MS spectra data as phaseollidin (1), L-maackiain (2), 2-(2',4'-dihidroxyphenyl)-5,6-methylenedioxy benzofuran (3), 8-demethyl-farrerol (4), liquiritigenin (5), genistein (6), 6-methylgenistein (7), 5-O-methyl genistein (8), 7,2',4'-trihydroxys-5-methoxy-isoflavanone (9), 7, 3', 4'-trihydroxy-isoflavanone (10), erythribyssin D (11), calycosin (12), trans-resveratrol (13), cis-resveratrol (14), stigmasterol (15), β-sitosterol (16). Among these, compounds 1-14 and 16 were isolated from this plant for the first time.
Chemical Fractionation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Sophora ; chemistry ; Spectrometry, Mass, Electrospray Ionization