Bone defect repair is a complicated process of connective tissue repair under the control of various cytokines.It is the ample blood supply which is a necessary condition to promote regeneration of fracture,especially bone defect and nonunion.Hypoxia inducible factor 1 can induce formation of neovascularization with perfect physiological function in the lesion of bone defect by regulating various gene expression,and thus provide nutritional support and favorable conditions to metabolism for different cells in the process of osteogenic differentiation and osteogenic activity to promote the healing of fracture.Both gene therapy and using hypoxia inducible factor 1 directly have an ability to promote neovascularization in the lesion of fracture.It is a hotspot at present that hypoxia inducible factor 1 in the field of bone tissue engineering is used to treat bone defect.Hypoxia inducible factor 1 transgenic therapy needs further research in the repairing process of bone defect.

Objective To seek the ancients’ method for acupuncture treatment of epigastric pain and explore the ancients’ regularities in point selection for acupuncture treatment of this disease to provide a reference and basis for guidance of modern clinical treatment. Method Search words related to epigastric pain were established. A database was set up. Frequency analysis and data mining technique-associated rules were used to investigate the use frequencies of different acupoints and meridians, and regularities in acupoint combination in ancient acupuncture treatment of epigastric pain. Result In ancient acupuncture treatment of epigastric pain, the points more frequently used and more accepted were Zhongwan (CV12), Zusanli (ST36), Neiguan (PC6), Shangwan (CV13), Geshu (BL17), Gongsun (SP4) and Jianli (CV11). Meridian points were the main ones. The points of meridians going through the epigastrium were often selected. Acupoint combination was mainly based on syndrome differentiation-based point selection plus symptomatic point selection.

Objective To investigate the application value of color Doppler ultrasonography in the diagnosis and treatment of uterine scar pregnancy. Methods The sonographic characteristics of 34 patients of uterine scar pregnancy were analyzed. The changes of blood flow and the resistance index (RI) were compared before and after conservative treatment. Meanwhile, the therapeutic effect was evaluated referencing to the changes of β-hCG in blood. Results Uterine scar pregnancies were classfied into three subtypes according to ultrasonic characteristics: Type Ⅰ: embryo sac (n=16 cases);Type Ⅱ: asymmetrical mass (n=14);Type Ⅲ: mixed type (embryo sac+asymmetrical mass, n=4). Peri-myometrium of the gastrula and the mass had abundant blood flow and lower RI before treatment, while rare blood flow and middle RI were found 4 weeks after treatment. There was statistical significance in the change of RI between before and four weeks, after treatment. Conclusion Uterine scar pregnancy has characteristic ultrasonogram. Color Doppler ultrasound may precise diagnose and localize uterine scar pregnancy, playing an important role in guiding the conservative treatment schedule, evaluating the therapeutic effect and following-up of these patients.

Objectives To learn about current public health education for undergraduate clinical students and to provide some references for developing suitable teaching way in the further.Methods Public health education for undergraduate clinical students in 11 medical colleges and universities and teachers' opinions on it were surveyed with the self-made questionnaire.Quantitative data were analyzed with descriptive statistic method.Results All the surveyed colleges and universities opened public health curriculum for undergraduate clinical students and 10 colleges and universities made public health course to be compulsory.The teaching contents were varied in different colleges and universities.Teachers who gave the public health courses proposed some suggestions on its reform.Conclusions Public health education for clinical students in different colleges and universities has both unity and diversity.It should develop new teaching model based on the training goal of public health education for undergraduate clinical students.

A novel hemizygous frameshift mutation in exon1of DAX-1 gene (993delC) was found in a patient with late-onset adrenal hypoplasia congenita and hypogonadotropic hypogonadism.This mutation led the stop codon to appear in advance of 59 amino acids.His mother and two sisters were the carriers of this hemizygous mutation while his father and brother were wild-type.After glucocorticoid hormone replacement therapy, the clinical symptom was improved, but the level of ACTH was not suppressed.

BACKGROUND: Vascular endothelial growth factor (VEGF) can promote angiogenesis, and has been extensively used in treatment of bone defect. However, few studies have addressed its isomer VEGF121. OBJECTIVE: To construct adenovirus vector carrying VEGF121-FLAG and humanized Renilla reniformis green fluorescent protein 1(hrGFP-1) and observe its expression in bone marrow stromal stem cells (BMSCs). METHODS: Using polymerase chain reaction technique, VEGF121 gene contained in the plasmid of pTG19T-VEGF121 was used to remove termination codon. NotI and Xho I restriction sites were added before and after gene sequence. Obtained gene subclone was moved onto pMD19-T plasmid. The pMD19-T-VEGF121 and pShuttle-CMV-IRES-hrGFP-1 plasmids underwent double enzymatic digestion. Small fragment and big fragment were retrieved utilizing gel. Subsequently, coupled reaction was conducted to complete the construction of shuttle plasmid. After measuring virus titer, BMSCs were transfected and the fluorescence intensity was observed under fluorescence microscope.RESULTS AND CONCLUSION: Recombinant adenovirus plasmid was successfully constructed by enzymatic digestion determination and gene sequence. Fluorescence microscope has shown that BMSCs transfected with recombinant adenovirus presented significantly green fluorescence expression. Thus, adenovirus vector carrying VEGF121-FLAG and hrGFP-1 gene can express in eukaryotic cells, which can be used for gene therapy for ischemic disease.

BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) can regulate the co-expression of various genes, and can induce angiogenesis with integrated physiological function.OBJECTIVE: To construct a novel adenoviral eukaryotic expression vector that can co-express mutant hypoxia-inducible factor-1 alpha (HIF-1a) target protein and humanized Renilla reniformis green fluorescent protein (hrGFP) reporter molecule under normoxic conditions.METHODS: The human HIF-1α gene carried by target gene donor plasmid pCMV6-XL5-HIF1α was sequenced and the site of restriction enzyme in above gene was analyzed. Site-directed mutagenesis of three amino acids including the 402 location, the 564 location, and the 803 location in gene coding region in HIF-1α were performed by polymerase chain reaction and sequencing was also done for monitoring mutation. The HIF-1α gene mutated correctly (HIF-1αmu) was coupled to adenoviral shuttle vector pShuttle-CMV-IRES- hrGFP-1. The recombinant adenovirus shuttle vector carrying HIF-1αmu gene was transferred to BJ5183-AD-1 electroporation competent cells after sequencing identification and Pme I restriction enzyme linearization.HIF-1αmu and hrGFP gene as well as hemeo-expression elements of hrGFP gene were reconstructed into adenoviral genome plasmids using homologous recombination mechanism in bacterium. Recombinants were obtained by Pac I restriction enzyme digestion and sequencing identification.RESULTS AND CONCLUSION: Amino acids including the 402 location, the 564 location and the 803 location in gene coding region in HIF-1α had become alanine after site-directed mutagenesis. Recombinant adenoviral expressing vector was successful as confirmed by restriction enzyme digestion and sequencing. These findings demonstrate that a novel recombinant adenoviral mutant eukaryotic expression vector pAd-HIF1αmu-IRES-hrGFP-1 was successfully constructed.

Objective:To explore the effects of exosomes derived from bone-marrow endothelial progenitor cells(EPCs-Exos)on the angiogenesis and collagen deposition in vitro,and to illustrate the possible mechanism for EPCs-Exos to accelerate the cutaneous deficiency repair.Methods:The endothelial progenitor cells (EPCs) were isolated, cultivated and identified;at the same time, the EPCs-Exos were also isolated and identified.The EPCs-Exos uptake in EPCs was observed and analyzed.16 rats were randomly divided into 4 groups, and then the models of skin defect were established, respectively.The equal volume of PBS and 50, 100, 150 mg·L-1 EPCs-Exos were injected into the area around skin defect of the rats in 4 groups.The wound closed sizes on the 0, 3rd, 7th and 14th days were measured, and Masson''s trichrome staining and CD31 immunofluorescence staining were performed on the 14th day for evaluating the tissue healing efficacy after EPCs-Exos treatment.In vitro,the mediums containing PBS and 50, 100, 150 mg·L-1 EPCs-Exos were used to culture the human umbilical vein endothelial cells (HUVECs), respectively.Scratch test and tubule formation assay were used to detect the migration and capillary network formation of HUVECs.At the same time, Western blotting was used for analyzing the expression level of angiogenesis related gene vascular endothelial growth factor A (VEGFA) in HUVECs.Results: The primary EPCs were isolated and identified successfully, and EPCs-Exos were purified and characterized.The CD31 immunofluorescence staining and double staining of DiL-ac-LDL and FITC-UEA-I of EPCs were positive.The electron microscope results showed that EPCs-Exos were nearly spheroidal, with the diameter about 40-100 nm.For the models of rat skin injury treated by EPCs-Exos, with the increasing of injection doses, the sizes of skin defect scar were gradually reduced, the degrees of scar healings were gradually increased,and the differences between various groups were statistically significant (P< 0.05).EPCs-Exos promoted the collagen maturity of healing skin in a dose-dependent manner;on the 14th day, the effect in 150 mg·L-1 EPCs-Exos group was the most significant.In vitro, EPCs-Exos promoted the migration and capillary network formation of HUVECs and increased the expression level of VEGFA;the migration rate,the net number of branches and the expression level of VEGFA in 150 mg·L-1 EPCs-Exos group were significantly higher than those in 50 mg·L-1 EPCs-Exos group and PBS group (P< 0.05).Conclusion: EPCs-Exos can promote the repair of traumatic skin defect of the rats by positively regulating the vascular endothelial cell function.

Objective To explore a new method for lacrimal passage irrigation after laser dacryocystoplasty surgery. Method One-hundred patients (104 eyes), which underwent laser dacryocystoplasty surgery combined with lacrimal drainage tube indwelling, were divided into two groups. In Group A (50 patients, 52 eyes), lacrimal passage irrigation was performed by traditional No.5 needle. While in Group B (50 patients, 52 eyes), it was done by No.7 blunt round needle designed by the researchers. Lacrimal passage irrigation was performed three times, each at the 3rd day, 1 week and 1 month after surgery. The silicone tube was removed 3 months after surgery and the treatment was evaluated. Results The total effective rate was 64% (32/50) in Group A and 92%(46/50) in Group B. The difference was statistically significant (χ2=18.537, P < 0.01). Conclusions The No.7 blunt round needle showed better effect when used for lacrimal passage irrigation after laser dacryocystoplasty surgery. It could improve the efficiency of lacrimal passage irrigation, thus decreasing irrigation times and reducing psychological pressure for the patients, which is worthy of clinical application.

A case of arginine vasopressin receptor 2 ( AV PR2 ) mutation in a boy with congenital nephrogenic diabetes insipidus was reported.Genomic DNA of the boy and his family members was extracted.The entire coding region of the AVPR2 gene were amplified by PCR.The amplified products were purified and sequenced.The results were compared with the normal one of the gene bank.The impact of the mutation on AVPR2 structure was discussed with respect to homology structure model.The analysis identified a T to G transition in exon 2 of the AVPR2 gene,resulting in substitution of leucine for arginine at amino acid residue 168.Furthermore,the patient′s mother and sister were heterozygous for this mutation,and the father was normol.