Raw264.7 cells were incubated with receptor activator of NF-kappa B ligand (RANKL) and α-melanocyte stimulating hormone(α-MSH) for6 d.The amount of osteoclast cells were counted by tartrate resistant acid phosphatase staining and the acid phosphatase activity was assayed.The expressions of 5 melanocortin receptors (MCR) in Raw264.7 cells were determined by RT-PCR.The results showed that the number of osteoclasts in RANKL +α-MSH group was significantly increased compared with RANKL group (P ＜ 0.05),but there was no osteoclast formation in α-MSH group.Compared with control group and α-MSH group,the acid phosphatase activities were significantly increased in RANKL group and α-MSH+RANKL group (P＜0.05).All five MCRs were expressed in the Raw264.7 cells shown by RT-PCR.These results suggest that α-MSH may promote osteoclasts formation through RANK signaling pathway.

Objective To establish a key technological system for spider fibroin gene code tandem connection , vector construction , prokaryotic expression and purification using genetic engineering in order to achieve MaSp 1 heterologous ex-pression in Escherichia coli and its separation and purification .Methods Isocaudarner ligation method was used to connect synthetic spider fibroin gene monomer code in tandem , and a recombinant clone concatemer was obtained .The identified recombinant clones were connected with prokaryotic expression vector pET 28a(+), and then transformed into E.coli BL21 (DE3).After being induced by IPTG for 6 hours, the expression product was identified by SDS-PAGE and Western blot-ting.Engineering bacteria were fermented in high density , and the obtained protein was purified through ammonium sulfate fractionation.Results and Conclusion The expression plasmids of MaSp1concatemers were successfully constructed , and the induced expression genetic engineering MaSp 1 protein was of the expected relative molecular mass .In addition, the pu-rity of the purified protein was above 80%.This study has developed crucial technologies for mass production of genetic en-gineering spider silk proteins .

Experimental investigations were conducted on the process of combustion and explosion vent in a 200 mm (diameter)×400 mm (length) vertical cylindrical vessel. When CH4-air mixture gases were used and the vent diameter was 55 mm,conditions of φ (equivalent ratio)=0.8, φ=1.0 and φ=1.3 and two ignition positions (at the cylinder center and bottom) were selected. The venting processes and the correlated factors are discussed in this paper.

Chinese patent orthopedic medicines feature complex components, mainly including desperate and toxic herbal pieces, narrow safety window, more clinical contraindications and frequent adverse drug reaction/events (ADR/ADE). To study the general safe medication regularity of Chinese patent orthopedic medicines, define key points in the medication education and ensure rational clinical medication, the authors took 80 types of commonly used Chinese patent orthopedic medicines as the study objects, collect 237 cases from 164 ADR/ADE documents through a system retrieval strategy, make a multidimensional literature analysis to determine the common risk factors for safe and rational medication of Chinese patent orthopedic medicines and establish an ADR/ADE prevention regularity. First, in the aspect of clinical symptoms, skin allergy is the most common ADR/ADE and closely related to the toxic ingredients, particularly accumulated liver or kidney damage caused by some drugs. Second, there are three time nodes in the ADR/ADE occurrence; The ADR/ADE occurred in 30 minutes is closely related to the idiosyncrasy; the ADR/ADE occurred between several months and half a year is related to the drug-induced liver and kidney damages; The most common ADR/ADE was observed within 7 days and predictable according to the pharmacological actions; Third, toxicity is an important factor in the occurrence of ADR/ADE of Chinese patent orthopedic medicines. Fourth, emphasis shall be given to the special medication factors, such as the combination with western medicines and Chinese herbal decoctions, overdose and long-course medication and self-medical therapy. In conclusion, the general ADR/ADE prevention regularity for Chinese patent orthopedic medicines was summarized to provide supports for clinicians in safe and rational medication and give the guidance for pharmacist in medication education.
Adult ; Aged ; Bone Diseases ; drug therapy ; Drug-Related Side Effects and Adverse Reactions ; etiology ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Nonprescription Drugs ; Orthopedics ; legislation & jurisprudence ; Young Adult

Objective To investigate the management of pregnancy and cardiovascular complications in women with Marian syndrome(MFS).Methods From October 1994 to September 2006, 30 patients with MFS undergoing cardiovascular surgery were studied retrospectively.Results In the labor of 46 offsprings given birth by 30 women,5 cases(11%)were performed elective cesarean section because of the existence of aortic complication,and 12(26%)were diagnosed as MFS.The gestation in two patients was terminated due to deterioration of aortic abnormalities during their third trimester,and they received surgical treatment with Bcntall procedure.Two developed acute aortic dissection during labor and post delivery respectively.With the manipulation of anticoagulation peripartum,one who had the implantation with mechanical prosthesis went through pregnancy and delivery uneventfully.The average duration between delivery and cardiovascular surgery was(15?9)years.Conclusions Vaginal delivery can be done safely in patients with the MFS who do not have or have mild cardiovascular system abnormalities,aortic dissection,or other important cardiac abnormalities,cesarean section should be the preferred method of delivery.Women with MFS are at increased risk for dissection and congestive heart failure during pregnancy and should be counseled before pregnancy about these risks,as well as the inheritance of the condition.

Objective To explore the effect of extracurricular activities based on specialist nursing clinic. Methods Thirty-four undergraduate nursing students of grade 2013 in the Capital Medical University were enrolled in the study. After learning of nursing theories for senile dementia, the extracurricular activities based onspecialist nursing clinicwere carried out. Before and after the second class activities, the caring ability inventory scale and self scale-rating for self-directed learning (SRSSDL) were used to assess 34 students. Result After the activities, students' caring ability and self-directed learning ability were statistically significantly improved (P<0.01). Conclusion The extracurricular activities based onspecialist nursing cliniccan stimulate the learning interest of nursing students and improve their self-directed learning ability and humanistic care ability.

BACKGROUND: Three-dimensional (3D) bioprinting technology has a huge potential in the tissue engineering field, which is expected to create simple tissue/organ analogues with good biological histocompatibility and biological functions by using living cells and biomaterials. OBJECTIVE: To analyze the characteristics of 3D bioprinting technology and all kinds of biomaterials, and to explore its application in the preparation of tissues/organs analogues. METHODS: Relevant articles published from 1998 to 2017 were searched in PubMed, Web of Science, MEDLINE, and WanFang databases. The keywords were "3D bioprinting, 3D bioprinting technology, biomaterial, tissue engineering" in English and Chinese, respectively. A total of 88 articles were initially searched and 47 eligible articles were finally reviewed in accordance with the inclusion and exclusion criteria. RESULTS AND CONCLUSION: 3D bioprinting techniques mainly include inkjet technique (thermal inkjet and piezoelectric inkjet), pressure-assisted technique, laser-assisted technique, and stereolithography technique (single-photon-based and two-photon-based). The bio-ink consists of living cells, natural polymers and synthetic polymers. 3D bioprinting has exhibited a huge potential in the manufacture of living cell-containing tissue/organ analogues. Despite the fact that it has been widely studied, currently used 3D bioprinting techniques can only be used to prepare relatively simple structures with simple biological functions. Research on the specific tissue/organ analogues with living cells are still in its infancy.

OBJECTIVETo evaluate the impact of a new CD44 variant on invasion of human breast cancer cell line MCF-7, and its possible mechanisms.

METHODSThe full length cDNA encoding CD44v17 was obtained from the total RNA isolated from the MCF-7/ADR cells by reverse transcript-polymerase chain reaction (RT-PCR) and subcloned into pMD19-T vector. The CD44v17 gene sequence and reading frame were confirmed by two restriction enzymes and nucleotide sequencing, and then inserted into the eukaryotic expression vector pcDNA3.1. The pcDNA3.1-CD44v17 was transfected into MCF-7 cells by Lipofectamine. The changes of MMP-2 and MMP-9 expression at gene and protein levels were detected by RT-PCR and gelatin zymography, respectively. The number of the cells through the artificial matrix membrane in every group was counted to compare the change of the invasive ability regulated by CD44 variant. The ERK and p-ERK were investigated by Western blotting to approach the molecular mechanisms of MMP-2 and MMP-9 expression regulated by CD44 variant.

RESULTSThe new gene sequence was successfully cloned into recombinant vector pcDNA3.1 and identified by the two restriction enzymes. It was confirmed that the reconstructed plasmid contained the sequence of CD44 gene variant which was composed of 1 to 4 exons, 16 to 17 exons, and 1 to 205 bases of 18 exons. The new gene sequence was sent to NCBI for publication and obtained the registered number FJ216964. The up-regulated levels of the CD44 gene mRNA and protein were respectively detected by RT-PCR and flow cytometry in MCF-7 cells transfected with pcDNA3.1-CD44v17. The invasiveness of the cells and the activity of MMP-2 and MMP-9 were clearly activated by hyaluronic acid (HA) treatment and blocked by CD44 neutralizing antibody. Pretreated MCF-7/CD44v17 cells with the neutralizing antibody against CD44 and the inhibitor of MAPKs signaling pathway strongly block the expression of p-ERK.

CONCLUSIONA new CD44 gene variant has been found in adriamycin-resistant human breast cancer MCF-7/ADR cells. The expression vector pcDNA3.1-CD44v17 has been cloned and constructed successfully. HA can be integrated with CD44 variant and then regulates the expression of MMP-2 and MMP-9, which increases the invasion ability of MCF-7 cells through the Ras/MAPK signaling pathway.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; metabolism ; Hyaluronic Acid ; pharmacology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Plasmids ; Protein Isoforms ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Signal Transduction ; Transfection

Objective To verify whether the periodic or continuous exposure to high glucose may have different effects on human umbilical vein endothelial cell (HUVEC)apoptosis, and to explore the effect of NF-κB pathway on apoptosis of HUVEC induced by high glucose using the RNAi adenovirus vector. Methods RNAi combinant adenovirus vector which targeted 1566 site of NF-κB p65 mRNA was constructed and the effect of p65 gene knockdown in HUVEC was detected by Western blot analysis. Then, the RNAi adenovirus was transducted to explore the role of NF-κB pathway on the regulation of apoptosis in HUVEC induced by high glucose. The apoptosis of HUVEC was tested by flow cytometry and TUNEL assay. Results High glucose could induce apoptosis of HUVEC. p65 protein expression of nuclear extracts was significantly increased in high glucose culture as compared to control group, but only slightly increased in NF-κB-specific knockdown group, which maintained at basal state. Compared with normal glucose group, the number of TUNEL-positive cells in high glucose group was significantly increased (25.81%±1.77% vs 8.20%±0.63%, P<0.05). The number of TUNEL-positive cells was decreased in 30.5 rmnol/L glucose plus Ad-1566 than that in 30.5 mmol/L glucose plus Ad-DEST (11.49%±0.92% vs 26.10%±0.98%, P<0.01). Flow cytometry and TUNEL assay showed that the apoptosis of human umbilical vein endothelial cells induced by high glucose was inhibited by the RNAi adenovirus. Conclusion High glucose induces apoptosis of HUVEC. Knockdown of NF-κB p65 may protect HUVEC from apoptosis by preventing high glucose-induced NF-κB nuclear translocation.

Experimental investigations were conducted on the process of combustion and explosion vent in a 200 mm (diameter) x 400 mm (length) vertical cylindrical vessel. When CH(4)-air mixture gases were used and the vent diameter was 55 mm, conditions of Phi (equivalent ratio)=0.8, Phi=1.0 and Phi=1.3 and two ignition positions (at the cylinder center and bottom) were selected. The venting processes and the correlated factors are discussed in this paper.