[Objective]To observe the effect of sciatic nerve and femoral nerve resection on heterotopic osteogenesis induced by recombined human bone morphogenetic protein-2(rhBMP-2) and discuss role of never innervation on bone regeneration and partial reasons for larger callus after bone fracture accompanying denervation.[Method]A total of 36 male ICR mice were divided into experimental group and control group at random.0.125mg rhBMP-2 /collagen composites were implanted into the right thigh muscle pouches after sciatic nerve and femoral nerve section in the experimental group and after sham operation in the control group.On the 7~(th),14~(th) and 21~(st) day after implantation,roentgenographic,biochemical,histological analyses and osteoclasts TRAP staining were performed to detect the effects of sciatic nerve and femoral nerve resection on bone growth initiated by rhBMP-2.[Result]On the 7~(th),14~(th) and 21~(st) day,wet weight of new bony tissues of the experimental group was obviously greater than that of the control group.Radiography showed range of bone formation in the experimental group was larger than that in the control,but density of new bone was lower than that of the latter.Biochemical detection showed value of AKP of the experimental group was significantly higher than that of the control on the 7~(th) day,while the content of Ca of the experimental group was higher than that of the control on the 14~(th) day and the content of Ca and P of the experimental group was lower than that of the control on the 21~(st) day.Histological observation showed area of new bone of the experimental group was greater than that of the control,but trabecular bone of the former was sparser than that of the latter.Image analysis of bony tissue showed the relative number of osteoclasts in the experimental group became higher on the 21~(st) day,while volume density and width of bone trabecula in the experimental group were lower than that of the control group.TRAP staining showed osteoclasts in the new bone of the experimental group were more activated than that of the control.[Conclusion]Nerve section results in enhancement of ability of bone regeneration by BMP in the early period,but trabecular bone becomes sparse in the middle and later period because of bone resorption by osteoclasts,which indicates nerve in nervation influences bone regemeration though direct and/or indirect way.

[Objective]To identify the expression of neurotrophic factors(NTFs)and their high affinity receptors(Trk)during the bone induction by recombined human bone morphogenetic protein-2(rhBMP-2)and discuss possible roles of NTFs on the osteoinductive activity of BMP.[Method]After a total of 36 ICR mice were divided into experimental group and control group at random,rhBMP-2/collagen sponge and collagen sponge were implanted into the right thigh muscle pouches of two groups respectively.Tissues in the implanted sites of two groups were removed on the 7~(th),14~(th) and 21~(st) day after implantation.Histological,immunohistochemical and RT-PCR analyses were performed to detect osteoinductive effect of rhBMP-2 and the expression of NTFs and Trk.[Result]RhBMP-2/collagen sponge displayed a potent ability inducing bone formation,while the expression of NTFs and Trk was observed in the course of osteoinduction by rhBMP-2.Especially on the 7~(th) day in the experimental group,NGF and TrkA positive immunostaining reached the peak in the stage of chondrogenesis,including chondroblasts,chondrocytes,hypertrophic chondrocytes and osteoblasts,then decreased in the number of positive cells and the intensity of immunostaining on the 14~(th) and 21~(st) day.BDNF was only observed in chondrocytes and cartilage matrix,and TrkB was expressed in chondroblasts and chondrocytes at the 7~(th) day after implantation.Expression of NT-3 was similar to that of NGF,while TrkC was expressed in chondroblasts and chondrocytes on day 7.Expression level of the mRNA of NTFs peaked at 7 days after the implantation,then decreased,which was nearly coincident with immunohistochemical results.[Conclusion]Many different kinds of cells are induced by rhBMP-2 express NTFs and Trk,which suggests that NTFs may play an important role in the osteogenesis initiated by BMP through direct and indirect pathways.

Objective To analyze the clinical application of high?throughput gene sequencing technolo?gy and STR in chromosome karyotype analysis of the villus tissues of spontaneous abortion. Methods In 27 ca?ses of spontaneous abortion after pregnancy,classic cell of villus tissues culture and chromosomal karyotype anal?ysis,and high ?throughput gene sequencing technology and STR were performed,and then compared the analysis results of two methods. Results ( 1) The successful rate of cell of villus tissues culture and chromosomal karyo?type analysis was 85%( 23/27) ,of high?throughput gene sequencing technology and STR was 96%( 26/27) ,and the difference was not significant( P>0. 05) ( 2) In the 4 cases that failed in karyotype analysis,there were 3 ca?ses showed abnormal chromosomal number variation( CNV) in high?throughput gene sequencing technology and STR. ( 3) Of the 23 cases,chorionic villus was successfully cultured in 10 cases,abnormal karyotypes were iden?tified in 13 cases,the positive rate was 57%. Of the 26 cases,high?throughput gene sequencing technology and STR was successfully checked in 3 cases,abnormal CNV were identified in 23 cases,the positive rate was 85%, the difference was significant(χ2=6.387,P<0.05). (4)The rates of chromosomal number abnormality were 52%( 12/23) and 50% ( 13/26) of karyotype analysis and chromosome aberration detection,respectively. In 10 cases of normal cell culture karyotype,there were 7 cases in the presence of micro deletion / micro repetition de?tected by high?throughput gene sequencing technology and STR. Conclusion The method of massively parallel sequencing in chromosome analysis,compared with the method of cell of villus tissues culture and chromosome a?nalysis,can be accurate and quick,and has high successful rate in detecting the chromosome of non aneuploid and deletion/duplication abnormality,which can be a good complementary and alternative method of the classic cell of villus tissues culture and chromosome karyotype analysis.

Objective To evaluate a new self-designed device to assist closed reduction of femoral shaft fracture.Methods From January 2012 to June 2014,18 cases of femoral shaft fracture received closed reduction and internal fixation with anterograde intramedullary nails on a popular fluoroscopic orthopedic table.The new self-designed distraction reduction device was used during the operation.The patients contained 14 males and 4 females,aged from 21 to 48 years (mean,33.6 years).Fracture AO classification was type 32A2 in 8 cases,32A3 in 5 cases,32B1 in 3 cases and 32B2 in 2 cases.Mean pre-operation time was 5.5 days (range,3-10 days).Results Operation time ranged from 85 to 130 minutes (mean,96 minutes).Intraoperative bleeding ranged from 120 to 180 ml (mean,150 ml).X-ray fluoroscopy time ranged from 20 to 40 seconds (mean,25 seconds).Frequency of nailing into the distal fracture end was 1 to 3 times (mean,1.5 times).One case had additional small incision for open reduction.Fourteen cases experienced external rotational deformity of the femur from 2° to 6° (mean,3.6°) after surgery,and four cases were free of rotational deformity.All patients were followed up for 6 to 20 months (mean,12.8 months).All patients had fracture healing in a period of 4.5 to 8.5 months (mean,5.5 months).No iatrogenic fracture,infections,nail breakage or other complications occurred.At the final follow-up,knee score according to the system of hospital for special surgery (HSS) was found to be excellent in 12 cases,good in 4,and fair in 2,giving an excellent-togood rate of 89％.Conclusion The new self-designed distraction device is an easy-to-operate instrument that is useful in assisting distraction and reduction of the femoral shaft fracture diffictlt to be fixed with routine methods as well as intramedullary nailing just on a popular fluoroscopic orthopedic table.

Objective To investigate the infectious status,etiological spectrum and epidemiological characteristics of rotavirus (group A/B/C),calicivirus (novovirus Ⅰ/Ⅱ,sapovirus),astrovirus and enteric adenovirus in diarrhea cases below 5 years old from 2008 to 2015 in Henan provinces.Methods Totally 2541 stool samples were collected from cases below 5 years old in four sentinel hospitals.All stool specimens were tested for group A rotavirus by double antibody sandwich ELISA method.G/P genotyping of group A rotavirus was determined by nested multiplex PCR.Viral RNA was extracted from all samples and rotavirus (group B/C),calicivirus,astrovirus and enteric adenovirus were detected by two-step reverse transcription-polymerase chain reation (RT-PCR)/PCR.Results One thousand four hundred and twenty-one out of 2 541 samples were positive with a total positive rate of 55 .9%,among which,102 were mixed infection.The isolation rate of rotavirus was 36.0% (914 samples)(group A:785 cases,group B:36 cases,group C:93 cases),calicivirus was 12.1 % (308 samples)(novovirus Ⅰ:64 cases,novovirusⅡ:193 cases,sapovirus:51 cases),astrovirus was 5 .9% (151 samples),enteric adenovirus was 1 .9%(48 samples).The group A rotavirus gene type combinations were composed mainly of G9P[8],G2P[4], G3P[8 ],G1P [8 ]and most cases were identified from September to November and March to May. Novovirus Ⅱ was predominant in calicivirus and most cases were identifed between March and May. Rotavirus or calicivirus infection was mainly among children aged 4—12 months or 3—5 years, respectively.Clinical manifestations included fever,diarrhea,vomiting,dehydration.Gender and region distributions differed according to the types of pathogen.Conclusions Group A rotavirus and novovirus Ⅱare the major viral pathogen in diarrhea cases younger than 5 years old in Henan province.Different viral infections exhibit extinct epidemiologic and clinical characteristics.

Objective To investigate the clinical value of evaluation of fetal cardiac function in congenital heart disease by brain natriuretic peptide (BNP) and velocity vector imaging (VVI). Methods Fetuses who came from Shenzhen Maternity & Child healthcare Hospital were divided into the congenital heart disease group and the control group. At the same time we collected amniotic fluid and assayed BNP concentration. Using the VVI software, the velocity, strain and strain rate of the global and segmental of the left ventricle were measured. Comparison and correlation were made between the two groups. Results There was significantly difference of BNP concentrations in amniotic fluid between two groups. The gestational age had significant positive correlation with BNP concentrations in disease group. The comparison of global velocity, strain and strain rate of left ventricle between the two groups showed significant differences. All of the left ventricular dynamic parameters in disease group were lower than those of the control group. Conclusions Compared with the control group, the disease group had a high level of BNP in amniotic fluid and a lower level of dynamic parameters of left ventricular. There was a positive correlation between BNP concentration and gestational age in disease group. So we can conclude that theBNP concentration can be a biological parameter for evaluating the latent impairments of fetal cardiac function.

To investigate the infection of Yersinia enterocolitica in Dengfeng City ,the strains were isolated from livestock and poultry .The strains were detected with biochemiological methods ,serological methods ,and virulence genes were detected with PCR .A total of 105 strains of Yersinia enterocolitica were classified from 1 285 stool samples ,the total isolation rate was 8 .17% .Among the total isolated strains ,17 strains were classified from dogs with a rate of 17 .35% and 35 strains from pigs with 13 .62% .Twelve strains were O ∶3 serotype (13 .48% ) ,12 strains were O ∶5(13 .48% ) ,and 14 strains were O ∶8 (15 .73% ) .Ail+ ,ystA+ ,yadA+ and virF+ accounted for 12 .36% ,and ystB+ accounted for 42 .70% .In conclusion ,the pigs and dogs were important animal hosts ,which may play the major role in humans'infection .

Objective To evaluate the clinical value of ultrasound breast imaging reporting and data system (BI‐RADS) in small solid breast masses with diameter ≤1 cm. Methods The ultrasound features of 230 solid breast masses with diameter ≤ 1 cm were described by ultrasound BI‐RADS, the relationship between ultrasound features, BI‐RADS final assesment and pathology were analyzed. Results Of these 230 masses, 72 (31 3.% ) were pathologically confirmed to be malignant and 158 (68 7.% ) to be benign. The ultrasound BI‐RADS features of mass shape, margin, orientation, posterior acoustic features, and microcalcificaition were significantly different between malignant and benign masses( P < 0.05). Irregular shape, noncircumscribed, nonparallel orientation, postrior shadowing, microcalcifications were regarded as malignant ultrasound features, their positive predictive values(PPV), sensitivity, specificity and accuracy for malignancy were 53 3.% -100%, 2 8.% -75 0.%, 82 3.% -100%, 69 6.% -80 9.%, respectively. One hundred and fifty‐two(66.1% ), 62(27 0.% ), 16(7 0.% ) masses were classified into grade 3, 4, and 5, respectively. The PPV for grade 3, 4 and 5 were 10 5.%, 64 5.%, 100% respectively. Among BI‐RADS grade 3 cases, 87 5.%malignant masses were intraductal carcinoma in situ and special type of invasive cancer, among pathological benign BI‐RADS grade 4 masses, 90 9.% were hyperplasia and intraductal papilloma. Conclusions In small breast masses with diameter ≤ 1 cm, due to the sensitivity of malignant signs are not high, the overlap between signs of benign and malignant lesions, pathological type and other factors, the positive predictive value of BI‐RADS grade 3 is higher than criteria of American College of Radiology, so BI‐RADS classification requires further detailed study.

Keratinocyte growth factor 2 (KGF-2) is a member of the FGF family that is mainly synthesized by mesenchymal cells and acta predominantly on epithelial cells in a paracrine manner. It is known to play an important role in fetal limb and lung development; skin wound healing and prostatic epithelial cell growth. The KGF-2 coding sequence were isolated from human kidney cDNA library, revealing that the Kgf-2 gene is also expressed in the kidney apparatus. Purified from prokaryotic E. coli cells, the effects of the recombinant KGF-2 protein in cultured keratinocyte were analyzed by using MTT assay and in situ TUNEL assay. Interestingly, results revealed that KGF-2 promoted keratinocyte cell growth by stimulating cell proliferation and attenuating cell apoptosis. These findings supported a few evidences that KGF-2 could contribute to alveolar epithelial cells against apoptosis. Cell migration assays for the first time revealed that KGF-2 could stimulate keratinocyte cell migration in vitro. In addition, in the pilot animal test, recombinant KGF-2 incorporated within the hydrogel dressing exhibited significantly stimulatory effect on cutaneous wound healing. These combined effects implicate a potential therapeutic application of human recombinant KGF-2 in the future.

Objective To observe the effect of lnng-term in vitro culture on the biological properties of adipose-derived stem cells(ADSCs)as seeding cells of tissue engineering.Methods The surface makers and apoptosis of primary and passaged human ADSCs were identified by flow cytometric analysis.Osteogenic differentiation of ADSCs at different passages were identified by alkaline phosphatase(ALP),Von Kossa staining and RT-PCR respectively.Results The surface marker expression of mesenchymal stem cells on ADSCs was high and did not change with passages of the cells.The early apoptosis rate of the cells was 1% to 2%,and increased insignificantly from passage one to passage nine.The osteogenic potential of ADSCs confirmed by ALP,Von Kossa staining and RT-PCR was maintained to as late as passage eight.Conclusion Since the biological properties of ADSCs are stable,they can be served as optimal seeding cells for tissue engineering and regenerative research.