Objective To construct the third-generation self-inactivating lentiviral vectors including BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a,then integrate them into the genome of mouse embryonic osteoblast cell,MC3T3-E1,and to explore the capability of osteogenic differentiation of individual bone morphogenetic control factor and the effective approaches to further improve the capability of osteogenic differentiation.Methods The plasmid vectors of gene expression including BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a were constructed,which were identified through enzyme cutting and further confirmed through sequencing.After packing pELNS-BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a,mouse embryonic osteoblast cell,MC3T3-E 1 was transfected,and the transfection efficiency was confirmed by GFP fluorescence imaging.The expression level of Runx2 mRNA and the transfection efficiency of individual bone morphogenetic control factor was detected by Real time PCR.Eight groups of MC3T3-E1 were dual-gene co-transfected,and the transfection efficiency was verified by GFP fluorescence imaging.ELISA was adopted to detect the expression level of BGP and ALP in MC3T3-E1culture supernatants;Real time PCR was adopted to detect the expression level of Runx2 mRNA;Western blot was adopted to detect the expression level of protein of BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a.Thus,the effectiveness of osteogenic differentiation of dual-gene co-transfection were evaluated.Results The recombination of lentiviruses,pELNS-BMP-2,pELNS-BMP-4,pELNS-BMP-6,pELNS-BMP-7,pELNS-BMP-9 and pELNS-Wnt3a were successfully constructed.MC3T3-E1 was successfully transfected.The expression levels of Runx2 mRNA were:BMP-2 ＞ BMP-4 ＞ BMP-9 ＞ BMP-7 ＞ Wnt3a ＞ BMP-6.Successful transfection of the dual-gene co-transfection of eight groups of MC3T3-E1 were verified by GFP fluorescence imaging.The expression level of Runx2 mRNA,the expression of BGP and ALP showed BMP-2 and BMP-7 co-transfection group was the most efficient in osteogenesis transfection.Western blot revealed thatthe protein expression of BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a increased after cotransfection of MC3T3-E1 by BMP-2 and BMP-7.Conclusion The third-generation lentiviral vector,pELNS can lead BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a into MC3T3-E1,mouse embryonic osteoblast cell,and stabilize its expression.Individual bone morphogenetic control factors can promote MC3T3-E1's differentiation to osteoblasts.The dual-gene co-transfection of BMP-2 and BMP-7 can effectively promote osteoblast conversion,which provides significant theoretical basis and technical support for remodeling of tissue engineering bone.

Objective To study the effect of different grades of gastric mucosa epithelial neoplasia tissues of Helicobacter pylori (Hp ) infection and the relationship between chemokine CXCL12 and its receptor CXCR4 expression and significance. Methods Based on a total of 138 cases of endoscopic specimens, there were normal gastric mucosa (NGM) in 32 cases, low grade intraepithelial neoplasia (LGIEN) in 35 cases, high grade intraepithelial neoplasia (HGIEN) in 36 cases, and gastric adenocarcinoma (GCA) in 35 cases. Hp infection was determined by 13C breath test and serum Hp antibodies,the double positive for Hp infection. CXCL12 and its receptor CXCR4 were detected by immunohistochemical method. Results The positive rates of CXCL12 and CXCR4 in LGIEN,HGIEN and GCA were higher than those in NGM:71.43%(25/35), 86.11%(31/36), 91.43%(32/35) vs. 25.00%(8/32);71.43%(25/35), 86.11%(31/36), 91.43%(32/35)vs. 28.12%(9/32), P<0.05;the positive rates of CXCL12 and CXCR4 in HGIEN and GCA were higher than those in LGIEN (P<0.05);the positive rates of CXCL12 and CXCR4 in HGIEN and GCA had no significant difference (P>0.05). The positive expression of CXCL12 and CXCR4 had no correlation with Hp infection (P>0.05). Conclusions The positive rates of CXCL12 and CXCR4 in LGIEN,HGIEN and GCA are higher than those in NGM.The positive rates of CXCL12 and CXCR4 in HGIEN and GCA are higher than those in LGIEN. The positive rates of CXCL12 and CXCR4 in HGIEN and GCA are similar. The positive expression of CXCL12 and CXCR4 have no correlation with Hp infection, it suggests that Hp infection may not play oncogenic role by CXCL12 and CXCR4 receptor expression.

Objective: In order to establish an effectively analytic method about the content of lead, arsenic, cadmium, mercury and copper. Methods:According to the standard addition method, matrix modifiers (nickle nitrate and diammonium orthphosphate) were added to sample and the contents of Pb, As, Cd and Cu were determined by graphite furnace atomic absorption spectrometry, reduce agent (acid tin bichloride was added to sample and the content of Hg was determined by cold vapour atomic absorption method. Results: The average recoveries of lead arsenic, cadmium, copper and mercury were Pb=97.5%, RSD = 2.75%; As=96.6%, RSD = 2.23%; Cd= 98.2% , RSD = 1.32% ;Cu=97.8%, RSD =1.19%;Hg= 98.8% , RSD = 1.85 , respectively. Conclusion: The method is sensible, accurate and reproducible. and applied for quality control of content of heavy metals in Qijidehuang Pills.

Objective To study the absorption features of isoimperatorin in intestine of rat.Methods Establish a single-pass intestinal perfusion model of rat,take phenolsulfonphthalein as a marker for the detection of isoimperatorin concentration from crude extracts of notopterygium,and observe the absorption features of isoimperatorin and its reference substance in intestine of rat.Results The content of isoimperatorin in crude extracts was (0.43±0.02)% (n=5).The absorption parameters of duodenum,jejunum,ileum,colon were (5.28±0.82),(4.47±0.56),(4.17±0.94),(4.32±0.68)×10-5 cm·s-1,respectively.There were no significant differences among them (P>0.05).Compared with the reference substance of isoimperatorin,crude extracts showed better absorption features.Conclusion Isoimperatorin from crude extracts have better characteristics of absorption.This study can provide theoretical basis for design of notopterygium oral formulation.

The cardiovascular diseases, which have the increasing incidence and high mortality rate, have brought a great impact on people′s lives and health.Being lipid inclusions with 30~150 nm in diameter, exosomes can be secreted by most cells in the body, and proteins, lipids, nucleic acids and other substances derived from those cells are wrapped inside.Exosomes play a variety of biological roles in the communication between cells, such as mediating immune response, inflammatory response, cell migration and differentiation.Research on exosomes has made significant progress, and the roles of exosomes in cardiovascular diseases have consequently attracted more and more attention in recent years.The communication of exosomes between cardiomyocytes and between heart and peripheral tissues has provided new strategies for the diagnosis and treatment of cardiovascular diseases.Application of exosomes in the diagnosis and treatment of cardiovascular diseases such as heart failure, viral myocarditis, Kawasaki disease and dilated cardiomyopathy is summarized in this review.

Objective To investigate the CT findings of histocyticnecrotizing lymphadenitis(HNL)in the neck.Methods CT data of 10 patients with pathologically confirmed HNL in the neck were retrospectively analyzed,7 males and 3 females,aged from 4 to 75 years old(median age 26 years old).Nine patients had plain CT scans and 5 of them had contrast scans.One case had only contrast CT scan.Results Totally,127 lymph nodes were identified in the neck,mainly located in the area of Ⅱ,Ⅲ,Ⅳ and Ⅴ.The maximum diameter of the involved lymph nodes ranged from 0.5-3.6 cm,1.3 cm in average.One hundred and eight lymph nodes were homogeneous and 8 were heterogeneous in plain CT images.Seventy nine lymph nodes had homogeneous enhancement and 27 had heterogeneous enhancement One hundred and ten lymph nodes had unclear margins and the surrounding fat was blurred.Conclusion CT findings of HNL of the neck are variable and non-specific.Clinical findings and laboratory examination may be helpful for the diagnosis and differential diagnosis.

OBJECTIVETo discuss the application of right-inferior-posterior "artery first" approach in laparoscopic pancreaticoduodenectomy.

METHODSClinical data of 17 patients who underwent laparoscopic pancreaticoduodenectomy through right-inferior-posterior "artery first" approach in our department from February 2014 to April 2015 were retrospectively analyzed. The operation began at the inferior flexure of duodenum. After entering the Toldt's space, the left renal vein (LRV) was revealed and the root of superior mesenteric artery (SMA) was exposed just above the LRV. SMA was dissected along its trunk till the horizontal part of duodenum.

RESULTSOf these 17 cases, adenocarcinoma of pancreatic head was observed in 5 cases, adenosquamous carcinoma in 2 cases, mucinous cycstic neoplasm in 1 case, adenocarcinoma of lower common bile duct in 4 cases, and duodenal papilla cancer in 5 cases. Fifteen cases were accomplished successfully with laparoscopy and 2 cases were converted to open approach. The average operating time was (320 ± 85) min and mean intraoperative blood loss was (305 ± 175) ml. The cutting margins were tumor negative in all the patients. The average number of harvested lymph node was 15.4 ± 6.5. Postoperative complication occurred in 5 cases. Two cases of bile leakage and 2 cases of pancreatic fistula were cured with conservative treatment. One case of delayed abdominal hemorrhage was resolved with reoperation.

CONCLUSIONRight-inferior-posterior "artery first" approach is safe and feasible in laparoscopic pancreaticoduodenectomy.

Adenocarcinoma ; Duodenal Neoplasms ; Duodenum ; Humans ; Laparoscopy ; Mesenteric Artery, Superior ; Operative Time ; Pancreas ; Pancreaticoduodenectomy ; Postoperative Complications ; Retrospective Studies

Objective: To investigate the clinical efficacy of recombinant human tumor necrosis factor-α receptor II: IgG Fc fusion protein combined with sodium hyaluronate in the treatment of rheumatoid arthritis (RA). Methods: Forty adult RA patients (12 males, 28 females) aged from 36 to 79 were selected in Department of Rheumatology of the Second People′s hospital of Datong from February 2016 to February 2018.The patients were randomly divided into the observation group and the control group according to the digital table, with 20 cases in each group.The observation group was treated by intra-articular injection of recombinant human tumor necrosis factor-α receptor II: IgG Fc fusion protein and sodium hyaluronate, and the control group was only treated with recombinant human tumor necrosis factor-α receptor II: IgG Fc fusion protein.The joint swelling, joint pain, joint mobility, morning stiffness, X-ray grade and total effective rate of the two groups were statistically analyzed before and after treatment. Results: Before treatment, there were no statistically significant differences in joint swelling, joint pain, joint mobility, morning stiffness, clinical score and X-ray grade between the two groups (all P>0.05). After treatment, the symptoms and signs of the two groups were obviously improved.The joint pain, joint pressure pain, joint activity, morning stiffness and clinical score in the observation group were (1.2±1.2)points, (0.8±0.7)points, (0.5±0.4)points, (0.7±0.7)points and (4.5±2.6)points, respetively, which were significantly better than those in the control group [(2.2±1.4)points, (1.5±0.9)points, (1.5±0.9)points, (1.5±0.6)points and (8.6±4.6)points, U=125, 111, 68, 89 and 97, all P<0.05]. The total effective rate of the observation group was 100% (20/20), while that of the control group was 75% (15/20), there was statistically significant difference between the two groups (χ²=14.10, P<0.01). Conclusion The injection of recombinant human tumor necrosis factor-α receptor II: IgG Fc fusion protein combined with sodium hyaluronate in the treatment of RA can effectively improve the clinical symptoms of RA patients, and it is more effective than single use.It is worthy of popularizing in clinic.

OBJECTIVETo study the micrometastasis distribution in liver tissue surrounding hepatocellular carcinoma (HCC), and provide reference for appropriate surgical safety margin.

METHODSThirty-six patients with HCC but without clinical metastasis underwent hepatectomy. Their specimens showing ample surgical margin were made into giant sections. Tumor micrometastasis in liver tissue around the primary tumor were examined microscopically. In each specimen, the surrounding tissue was divided into proximal(p) and distal(d) areas. In either area, three lines of demarcation 0.5 cm, 1.0 cm, and 2.0 cm away from the margin of the primary tumor were designated as L(0.5), L(1.0) and L(2.0). Therefore, the surrounding tissue was divided into six zones - Z(p0.5), Z(p1.0), Z(p2.0) and Z(d0.5), Z(d1.0), Z(d2.0). The maximum micrometastasis spread distance (MMSD) and density (D(p0.5), D(p1.0), D(p2.0) and D(d0.5), D(d1.0), D(d2.0)) in each zone were analyzed after search for micrometastasis in the giant sections.

RESULTS72.5% (111/153) micrometastases were found in form of microscopic tumor emboli. Their spread distance could be up to 6.1 cm. In 66.7% (24/36) specimens, micrometastases were found in the surrounding tissue. In 91.7% (22/24) of them, the distal MMSD was less than 3 cm. The proximal MMSD was less than 1.5 cm in 92.3% (12/13). The comparison of micrometastasis density in the different zones were D(d0.5) > D(d1.0) > D(d 2.0); D(p0.5) > D(p1.0) > D(p2.0); D(d1.0) > D(p1.0); D(d2.0) > D(p2.0) with significant differences.

CONCLUSION(1) Micrometastases of HCC exist mainly in form of microscopic tumor emboli, (2) The longer the distance from the primary focus, the lower the micrometastasis incidence, (3) In zones more than 0.5 cm away from the primary focus, tumor micrometastasis incidence is significantly lower in the proximal zones than that in the distal zones and (4) For HCC patients without clinical metastasis, a surgical margin of 3 cm wide in the distal area and 1.5 cm wide in the proximal area may reduce the rate of postoperative recurrence.

Carcinoma, Hepatocellular ; secondary ; surgery ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; pathology ; surgery ; Neoplasm Metastasis ; Neoplasm Recurrence, Local

Objective To investigate clinical and histopathological features of lichen planopilaris (LPP).Methods The clinical and histopathological findings in 3 cases of LPP were analyzed.Results All the 3 patients were female,and their average age was 49 years.One patient presented with large-area patchy hair loss on the frontal,parietal and occipital region,and 2 patients presented with irregular patchy hair loss on the scalp and skin atrophy.Besides the hair loss,the eyebrows and axillary hairs also lost in 1 patient.Histopathological examination showed liquefaction degeneration of basal cells in the walls of hair follicles and infiltration of lymphocytes.Infiltration of a small number of lymphocytes was also observed around blood vessels and appendages.Conclusions LPP may only affect the scalp,or involve the other sites all over the body.LPP commonly manifests as patchy hair loss on the scalp and skin atrophy,and is pathologically characterized by liquefaction degeneration of basal cells in hair follicles and infiltration of lymphocytes.