Objective To compare the effects on blood specimen collection from neonatal heel by oblique puncture and vertical puncture methods,to increase the efficiency of puncturing in blood collection.Methods Two hundred eutocia,full-term infants were randomly divided into experimental group (oblique puncture) and control group( vertical puncture) by stochastically ballot,100 samples in each group,and the blood collection rate in two groups was compared.Results In the same condition,both experiment group (87 cases) and control group (56 cases) have higher efficiency in blood collection,but the experiment group is notably easier than control group (x2 =8.76,P＜0.01),however there is no significant difference in pain (t =0.55,P =0.58).Further,we observed the response of neonatal heel and found that both methods will not induce inflammation,but the peidnoma was significantly decreased in experiment group compared to control group ( x2 =9.26,P ＜ 0.01 ).The experiment group was significant high than control group.Conclusions Oblique puncture method is conformance to care program,and can significantly decrease the suffering of newborns,and increase the efficiency.

Objective:To explore the methods and ideas for developing pharmaceutical care in clinical practice. Methods: The pharmaceutical care performed by clinical pharmacists and the therapeutic scheme assisted by clinical pharmacists for one patient with adult purulent meningitis were analyzed retrospectively. Results and Conclusion:Through selection of anti-infective agents, treatment of adverse drug reactions and assessment of patients’ economic capacity,clinical pharmacists help provide reasonable medication to im-prove therapeutic efficacy, safety and economy.

Objective To evaluate the CT findings of pancreatic carcinoid tumors.Methods The CT imaging data of five patients with pancreatic carcinoid tumors confirmed by pathology were retrospectively analyzed.Results The tumors ranged in maximum diameter from 2.0 to 11.0 cm with a mean of 6.4 cm. On unenhanced CT,the tumors were slightly hypodense relative to the pancreatic parenchyma,homogenous in 2 cases,and heterogenous in 3 cases.One tumor showed calcification.After contrast material injection, the solid component of the tumor showed marked heterogenous enhancement on the arterial phase scanning in 3 cases,and mild heterogenous enhancement in 2 cases.The degree of tumor enhancement was less intense than the surrounding pancreatic parenchyma due to necrosis of various degree,which led to the cystic appearance of the tumor in 1 ease.On the portal phase scanning,all tumors showed marked enhancement similar to that of the pancreatic parenchyma.On the delayed phase scanning,the degree of enhancement was more intense than the surrounding pancreatic parenchyma in 1 case.Liver metastases with retroperitoneal lymphadenopathy and peripancreatic vessels invasion were seen in 1 case.No dilatation of the biliary tract or pancreatic duct was present.Conclusion The CT features of pancreatic carcinoid tumors included infrequent dilatation of the biliary tract or pancreatic duct and unusual vascular involvement,calcification within the mass,marked enhancement similar to that of the surrounding pancreatic parenchyma during the portal phase scanning and more intense during the delayed phase scanning.

Objective To investigate the mechanism of klotho reversing the resistance of breast cancer to paclitaxel in MCF-7/PTX cells.Methods The Klotho expression in MCF-7 and MCF-7/PTX cells was detected by Western blot.The effects of Klotho on paclitaxel resistance in MCF-7/PTX cells was measured by MTT assay.The effects of Klotho and 3-methyladenine (3-MA) on proliferation and expression of Beclin1 in MCF-7/PTX cells were detected by MTT and Western blot assay,respectively.Results The expression of Klotho in MCF-7/PTX cells was decreased compared with MCF-7 cells.Klotho could sensitize MCF-7/PTX cells to paclitaxel.The expression of Beclin1 in MCF-7/PTX cells was higher than that in MCF-7 cells.Klotho and 3-MA could decrease the expression of Beclin1 in MCF-7/PTX cells,and the effects of Klotho on paclitaxel resistance in MCF-7/PTX cells was similar to that of 3-MA.Conclusion Paclitaxel resistance in breast cancer cells is related to expression of the Klotho which can reverse the resistance of breast cancer to paclitaxel by inhibiting autophagy.

Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Female ; Fractures, Comminuted ; surgery ; Hip Fractures ; surgery ; Hip Prosthesis ; Humans ; Male

Objective:To analyze the factors which may influence postoperative early ambulation in patients with fresh fracture so as to further optimize the perioperative protocol based on the concept of enhanced recovery after surgery (ERAS).Methods:A retrospective analysis was conducted of the patients who had been treated for a single fresh fracture at the extremity, pelvis or acetabulum from May 2019 to July 2019. Collected were the data concerning basic features of patients, perioperative ERAS management and surgery, as well as early ambulation on the day of surgery or the first day after surgery. The patients were divided into an early ambulation group and a non-early ambulation group according to the time of ambulation. Statistical analyses were performed of the relationships between early ambulation and 20 potential factors concerning basic features of patients, perioperative ERAS management and surgery. Logistic correlation analysis was performed to identify risk factors for postoperative early ambulation.Results:A total of 306 patients were included, including 96 upper limb, 203 lower limb, 5 acetabular and 2 pelvic fractures. Of them, 150 ambulaed from bed on the day of surgery or the first day after surgery while 156 did not. Significant differences were observed between the 2 groups in fracture site, intake of carbohydrate liquids the night before surgery and the day of surgery, catheter indwelling, intraoperative liquid transfusion volume and postoperative complications ( P < 0.05). Logistic correlation analysis of the relationship between the above factors and postoperative early ambulation found that fracture site and intraoperative liquid transfusion volume were significantly correlated with postoperative early ambulation ( P < 0.05). Conclusions:About half of the patients with a single fresh fracture may ambulate early after surgery. Fracture site and intraoperative liquid transfusion volume may be significantly correlated with postoperative early ambulation.

OBJECTIVETo evaluate the correlation of 64-multidetector-row CT (64MDCT) perfusion imaging with microvessel density(MVD) and vascular endothelial growth factor(VEGF) in colorectal carcinoma.

METHODS64MDCT perfusion imaging was performed in 33 patients with pathologically verified colorectal carcinoma. Time-density curves (TDC) were created from the region of interest (ROI) drawn over the tumor, target artery and vein by 64MDCT with perfusion functional software. The individual perfusion maps generated were for blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability-surface area product (PS). MVD and VEGF expression of surgical specimens were examined by immunohistochemical staining with anti-CD34, anti-VEGF monoclonal antibody. MVD and VEGF were compared among the different types of TDC in colorectal carcinoma. The correlation of CT perfusion parameters with MVD and VEGF was also examined.

RESULTSTDC of colorectal carcinoma was divided into five types according to their shapes. MVD in the colorectal carcinoma was 22.61+/-9.01. VEGF staining was found in 25 of 29 tumors (86.2%). The score of VEGF expression was 4.15+/-1.09. No significant differences of MVD and VEGF expression among TDC types were found (F=2.59, 1.11, P>0.05). There were also no correlations of MVD and VEGF expression with any dynamic CT parameters (P>0.05).

CONCLUSION64MDCT perfusion imaging, MVD and VEGF may reflect angiogenic activity, but no significant correlations are found among them.

Adult ; Aged ; Colorectal Neoplasms ; blood supply ; diagnostic imaging ; Female ; Humans ; Male ; Microvessels ; Middle Aged ; Neovascularization, Pathologic ; Tomography, Spiral Computed ; methods ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVEUtilizing the hypoxia inducible factor 1/hypoxia reaction element (HIF-1/ HRE) gene regulation system to construct antisense vascular endothelial growth factor (VEGF165) cDNA eukaryotic expression vector promoted by HRE, and investigate its targeted inhibiting VEGF expression of osteosarcoma cells in hypoxia environment.

METHODSEukaryotic expression plasmid with HRE promoter was constructed containing luciferase reporter gene and antisense VEGF165 cDNA by using PCR and recombinant DNA techniques. The recombinant vectors were transfected into osteosarcoma cells with lipofectin method. Hypoxia-inducible reporter gene expression was determined by liquid scintillation analyzer and the expression of VEGF protein was detected by ELISA method.

RESULTSThe eukaryotic expression plasmid containing antisense VEGF165 and luciferase promoted by HRE was constructed successfully. After being transferred into MG63 cells, luciferase expression was increased 3.5 x 10(2) times and VEGF protein expression decreased 45% under hypoxia condition.

CONCLUSIONAntisense VEGF165 cDNA expression, efficiently realized by HRE promoter under hypoxia condition, provides an experimental basis for targeted antiangiogenesis of tumors.

Bone Neoplasms ; metabolism ; pathology ; Cell Hypoxia ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1 ; genetics ; Luciferases ; genetics ; metabolism ; Oligodeoxyribonucleotides, Antisense ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Promoter Regions, Genetic ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the influence of CD44 a cell-matrix adhesion molecule on the proliferation, adhesiveness and invasiveness of osteosarcoma cell lines, in order to investigate the growth and invasion mechanism of osteosarcoma.

METHODSThree osteosarcoma cell lines MG-63, HOS and U2-OS were routinely cultured. Flow cytometry and Western blot analysis were used for detecting the positive rates and relative amount of CD44 protein in the three cell lines. RT-PCR method was also used to compare the differences in the expression of CD44 mRNA among the 3 cell lines. Then, MTT method, adhesion detection, and Microcon-migration assay were used to detect the changes of the cells' proliferation rate, adhesive and invasive abilities after blocking the role of CD44 by using a special neutralizing antibody.

RESULTSThe results of flow cytometry showed that the percentage of CD44 positive cells were both over 99% in HOS and U2-OS, while that in MG-63 was only (2.10 +/- 0.46)%. The average fluorescence density of CD44 in HOS was significantly higher than in U2-OS. Western blot also showed that the relative content of CD44 protein in HOS was notably higher than that in U2-OS, while CD44 was negatively expressed in MG-63. The expression of CD44 mRNA was significantly lower in MG-63 than in both HOS and U2-OS, which were consistent with the expression of CD44 protein. The proliferation rates and adhesive abilities of MG-63 and HOS have no significant difference, but both were significantly higher than that of U2-OS. The invasive abilities of HOS was dramatically higher than MG-63 and U2-OS. After the role of CD44 was blocked by anti-CD44 neutralizing antibody, the proliferation rates of the 3 cell lines did not change significantly. While the HOS and MG-63 adhesion indices decreased dramatically (P < 0.05), the invasive abilities of HOS and U2-OS also decreased notably (P < 0.01).

CONCLUSIONSCD44 could promote the adhesiveness and invasiveness of osteosarcoma cell line HOS. CD44 may take part in promoting the process of U2-OS invasion and the adhesion of MG-63. On the other hand, CD44 could not affect the osteosarcoma cell proliferation rates.

Bone Neoplasms ; metabolism ; pathology ; Cell Adhesion ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Hyaluronan Receptors ; biosynthesis ; genetics ; physiology ; Neoplasm Invasiveness ; Osteosarcoma ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the effect of VEGF expression in osteosarcoma cell line and the target killing effect of HSV1-TK/GCV system on transfected osteosarcoma cells under hypoxia conditions.

METHODSEukaryotic expression plasmid with HRE promoter was constructed to express the antisense VEGF165 cDNA and Hygromycin phospho-transferase-thymidine kinase (HyTK) fusion gene. The recombinant vectors were then transfected into osteosarcoma cell line MG63 with lipofectin mediated gene transfer methods. PCR and RT-PCR were used to confirm the presence and expression of TK gene. The sensitivity of transfected cells to GCV and "bystander effect (BSE)" of HSV1-TK/GCV system under normoxia or hypoxia conditions were measured by MTT assay and mixed co-culture experiment. The expression of VEGF protein was detected by ELISA under hypoxia condition. Cell cycle phase distribution was determined by flow cytometry. In addition, electromicroscopy was used to document ultrastructural alterations.

RESULTSThe eukaryotic expression vector pBI-HRE-AsVEGF165 -HyTK was constructed successfully. The transfected cell line MG63TV was established and confirmed by PCR and RT-PCR of the presence of transgene and its mRNA expression. GCV was toxic to transfected cells in a concentration-dependent manner. The sensitivity to GCV toxicity was 100 times higher under hypoxia condition than that under normoxic condition. The mixed culture experiments showed that the "bystander effect" was enhanced significantly under hypoxia condition. VEGF expression of transgene cells under hypoxia condition decreased 50% compared to that of normal condition. Under hypoxia and GCV, DNA synthesis of MG63TV cells was inhibited along with an increase of cells at G0 approximately G1 phase, apoptosis and necrosis.

CONCLUSIONSAntisense VEGF expression driven by HRE promoter in combination with hypoxia can provide a target inhibition of VEGF expression in human osteosarcoma cells, with an enhanced selective killing effect and BSE of the HSV-TK/GCV system. The double-gene co-expression system in study provides experimental basis for therapy against osteosarcoma by a synchronous antiangiogenic and suicide gene approach.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Bystander Effect ; Cell Hypoxia ; Cell Line, Tumor ; Cell Proliferation ; DNA, Neoplasm ; biosynthesis ; Ganciclovir ; pharmacology ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1 ; genetics ; Oligodeoxyribonucleotides, Antisense ; Osteosarcoma ; metabolism ; pathology ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Thymidine Kinase ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism