Objective To design an effective universal DNA vaccine for influenza A virus and detect its immune efficacy in mice.Methods The DNA sequence containing the M2(3M2e)extracellular domain of H1N1,H3N2 and H9N2 subtypes of influenza A virus was synthesized and cloned into vector pVAX1 to construct eukaryotic expression plasmid pVAX1-3M2e. The plasmid pVAX1-3M2e was mixed with cell penetrating peptides(CPPs)RVG9dR,Protamine,and RVG9dR +Protamine peptide complex(1∶1)at different mass fractions(1∶0. 5,1∶1,1∶2,1∶4,and 1∶8),separately,and gel retardation was performed to determine the proper binding of DNA and CPPs. The delivery efficiency of the CPPs was detected by cell fluorescence assay at cellular level(293T cells). Male BALB/c mice were immunized intramuscularly with plasmid pVAX1-3M2e delivered by RVG9dR + Protamine(two doses:25 μg pVAX1-3M2e + 500 μg polypeptide complex,50 μg pVAX1-3M2e + 1 000 μg polypeptide complex,8 mice in each group)to detect the cross-protection effect of universal influenza A virus DNA vaccine against three subtypes of influenza viruses(H1N1,H3N2 and H9N2).Results The optimum binding ratios of DNA to RVG9dR,Protamine,and RVG9dR + Protamine were 1∶2,1∶2,and 1∶1,respectively. RVG9dR + Protamine had superior delivery efficiency to plasmid pVAX1-3M2e at the mass fraction of 1∶20. The protective rates of 50 μg pVAX1-3M2e + 1 000 μg polypeptide complex against three subtypes of influenza virus in mice were all 100%.Conclusion The plasmid pVAX1-3M2e delivered by RVG9dR + Protamine polypeptide complex provides cross protection against three subtypes of influenza A virus in mice.

Objective To compare the efficacy and safety of endoscopy and laprotomy for gastric stromal tumor,and to evaluate the clinical value of endoscopy.Methods A retrospective analysis was made based on the clinical data of 142 patients with gastric stromal tumor(diameter＜3 cm with clear boundary),with 83 patients who underwent endoscopic resection assigned to the endoscopy group,and 59 patients who underwent laprotomic resection to the operation group.The clinical and pathological data in both groups were analysed and the safety,perioperative situation and prognosis of two groups were compared.Results Compared with the laprotomy group,the endoscopy group required the same time,[(88.25±12.18) min VS (90.15±10.25) min,P=0.367],yielded to less intraoperative blood loss[(42.15±22.13) ml VS (140.55±35.66) ml,P＜0.01],early postoperative feeding[(2.30± 1.20) d VS (5.40± 1.28) d,P＜0.01],and shorter postoperative hospital stay [(5.43± 1.78) d VS (9.42± 1.65) d,P＜0.01].Follow-up was performed in two groups for 5-55 months (median,30 months),with no recurrence or metastasis.Conclusion Compared with surgery,endoscopic therapy is safe and effective for the gastric stromal tumor,whose diameter is less than 3 cm with clear boundary.

OBJECTIVE To obser ve t he histopathological features and immunohistochemical expression of IgG4 in nasal granulomatosis with polyangiitis(GPA) and to study the relationship between GPA and IgG4-related disease(IgG4-RD). METHODS A total of 12 nasal GPA patients were collected from the Department of Pathology, Beijing TongRen Hospital during January 2012 to December 2016. EnVision immunohistochemical technique was used to detect the expressions of CD38, CD138, IgG and IgG4. RESULTS Nine cases were found pulmonary abnormalities. Elevated titer of anti-neutrophil cytoplasmic antibody was presented in 8 cases in this study. On histopathologic evaluation, 8 of 12 cases were graded as pathologic score 3, one as score 2, and the remaining 3 as score 1. Eight cases showed increased number of IgG4+ cells higher than 10/HPF, 2 of which showed the ratio of IgG4+/IgG cells were more than 40％. CONCLUSION IgG4+ cells can be increased in biopsies of nasal GPA. Since the morphologic and clinical manifestations of GPA and IgG4-RD may overlap, it could be a significant diagnostic pitfall in the differential diagnosis of these two entities. The diagnosis of GPA should combine clinical manifestations, imaging findings, serological examinations and pathological features.

BACKGROUND: Knee joint trauma results in knee instability and movement track changes, which lead to the degradation of articular cartilages. The main component of articular cartilages, such as chondrocytes, collagen and proteoglycan, are also change during this process.OBJECTIVE: To compare the content changes of collagen, proteoglycan and the apoptosis of chondrocytes on two model groups of knee joint instability, additionally, to investigate the occurrence mechanism of early articular cartilages changes in rabbits with knee joint instability.METHODS: A total of 50 white rabbits were randomly divided into anterior cruciate ligament transection group, medial collateral ligament transection + meniscectomy group and normal control group. The specimens were harvested at weeks 1, 2, 3, 4, and 5 after operation. The osteocartilaginous section of the medial femur condyles were observed by gross observation, histological examination, and Markin scores. The contents of proteoglycan and collagen fiber were also measured.RESULTS AND CONCLUSION: Chondrocytes apoptosis were found with certain degrees at different stages of knee joint instability models. The Mankin score of the anterior cruciate ligament transection group was greater than the medial collateral ligament transection + meniscectomy group at each time point except the 1<'st>week (P ＜ 0.05), which was notably greater in the 5~(th) week. At weeks 3 and 4 after operation, the chondrocytes degenerated with proteoglycan lost in the anterior cruciate ligament transection group, and the chondrocytes degeneration was predominant at 5th week. The changes were more seriously in the medial collateral ligament transection + meniscectomy group. Meantime, the collagen fiber content of two experimental groups were less than the normal control group at Weeks 3, 4, 5 after operation (P < 0.05). The results demonstrated that at early knee joint instability, the articular cartilages degenerated extent is related to the number of chondrocytes, the collagen network disrupts and lost of proteoglycan. During chondrocytes degeneration, cartilage repair simultaneously occurred.

Objective To discuss the technical factors of extrapleural locating in CT-guided percutaneous pulmonary biopsy. Methods 137 cases undervent CT-guided percutaneous pulmonary biopsy were retrospectively analyzed. The technical methods of operation,points and points for attention were analyzed. Results In this group the success rate of data manipulation was 98.5%. No serious complications of the operation were occured. Scan completed within 40-story biopsy rate was 71.5%. The needle into the target rate for the first time was 93.4%. Conclusion CT-guided biopsy could increase the detection rate of difficult lesions and patient satisfaction rate paying attention on thinking and practice in the preoperative preparation and intraoperative specific operations.

Objective To assess the effect of intraarterial washout with UW solution and tanshinone Ⅱ A for skeletal muscle preservation during ischemia and reperfusion.Methods Ischemic rat limbs were perfused with UW solution or UW solution plus tanshinone Ⅱ A (0.05,0.1 and 0.2 mg/ml) for 0.5 h before reperfusion; controls received no perfusion.Serum CPK,LDH,and AST were measured pre-ischemia and after reperfusion (2 h,4 h,and 6 h).Muscle water content,MDA,SOD,ATPase were assessed pre-reperfusion and after 6 h reperfusion.ICAM-1 was detected after 6 h reperfusion.Results Intraarterial washout with UW and UW + T could inhibit the express of ICAM-1 in skeletal muscle.The serum levels of CPK,AST,and LDH in UW group were significant lower than those in control group after 2 h of reperfusion,but no difference was observed between UW group and control group after 4 h and 6 h of reperfusion.After 4 h of ischemia,there were significant differences in water content,MDA,SOD,and ATPase in skeletal muscle between UW group and control group,but no difference was found after 6 h of reperfusion.In contrast,all parameters of laboratory test and biochemical analyzing in UW + T(0.05,0.10,and 0.20 mg/ml) groups were significantly different from those in control group at corresponding reperfusion period.Conclusion Intraarterial washout with UW solution is effective in preserving skeletal muscle integrity against I/R insult.Tanshinone Ⅱ A as a beneficial adjunct to UW solution improves the protective effect of UW solution for ischemic skeletal muscle.To better preserve ischemic skeletal muscle,an appropriate dose of tanshinone Ⅱ A (0.1 or 0.2 mg/ml)added to UW solution is required.

OBJECTIVE To investigate the clinicopathological features of ossifying fibroma (OF) and fibrous dysplasia (FD) , and to evaluate the role of immunohistochemistry in differential diagnosis. METHODS Clinical presentation and morphological study of 40 cases with OF and 26 cases with FD were reviewed. Immunohistochemistry stains for PCNA, Ki-67, Laminin and CK 19 were performed in specimens of 10 cases with FD and 26 cases with OF. RESULTS The average age of OF was 22.2 years and the ratio of male and female was 2.33:1. The average age of FD was 17.2 years and the ratio of male and female is about1:1. The average age of the multibone type of FD was 11.2 years and the ratio of male and female is 1:2. Most of OF occurred in ethmoid sinus (75 %) and 70 % of the patients presented snuffle and hyposmia, while FD inclineed to arise from sphenoid (58 %) and 80.8 %of patients showed eyesight dropping and exophthalmos. The immunohistochemistry stains PCNA was positive in 19 cases (73.1 %) of OF and negative in 10 cases of FD. Ki-67, laminin, CK19 were of little significance. CONCLUSION The gender, age distribution, clinical and pathological features of OF and FD are different from each other. Immunohistochemistry staining for PCNA is of great value in their differential diagnosis.

OBJECTIVE To investigate the relationship of obstructive sleep apnea hypopnea syndrome (OSAHS) and hyperglycemia by analyzing the level of blood-fasting sugar (BFS) in patients with OSAHS. METHODS The data of BFS and AHI in patients with OSAHS (n=300) and the data of BFS of controls (n=225) were collected. The difference of high blood glucose in the OSAHS and the controls.and the difference of AHI between OSAHS patients with high blood glucose and OSAHS patients with normal blood glucose was analyzed. RESULTS There were 97 patients (32.3 %) with high BFS in 300 OSAHS patients, but only 14 cases (6.2 %) with high BFS in 225 controls. There was a significant difference between two groups. The average of AHI in OSAHS patients with high BFS was 67.8/h, and average of AHI in OSAHS patients with normal BFS was 38.4/h. There was a significant difference between the two groups. CONCLUSION There is a close correlation of OSAHS and hyperglycemia.

Objective: To observe the influences of infusing small volume of 7.5% NaCl solution (HS) in vivo,and the different volume ratio of plasm of rich platelets (PRP) to HS or sucrose in vitro on the function of platelet. Method:Twenty-nine patients undergoing middle or small operation were divided randomly into HS group and normal saline group(NS). Blood samples were taken before infusion and,5,30,60 and 120 min after infusion,to measure hematocrit (HCT),platelets (PLT) count, maximum aggression rate (MAR),thromboxance B_2(TXB_2),granule membrane protein-140(GMP-140) and platelet factor 3 activety (PF_(3a)T). In addition,fifteen blood samples of blood donors were taken,and divided control group,HS group,sucrose group and NS group. The volume ratio in each group were arranged in 1/25,2/25,4/25 and 8/25. Osmotic concentration of 1/25 was equivalent to that induced with small volume hypertonic saline infusion. Result: Compare with NS group,the HCT and PLT count were decreased after infusion of small volume of HS. When the volume ratio ≥2/25, all measure values of the MAR, TXB_2, GMP-140 and PF_(3a)T were decreased in vitro,and profoundly inhibited following osmolarity increase of HS. Conclusion;Small volume of HS may be safely used,but severe inhibition of functions of platelet may occur when the infusing volume of HS exceeded more than twice of small volume of HS.

BACKGROUND: Myogenetic seed cells are the fundamental for construction of tissue-engineered smooth muscle, and optimizing the amplification of seed cells is the key in further clinical application. OBJECTIVE: To analyze the myogenetic potential of rabbit adipose-derived stromal cells induced with the combination of MyoD, transforming growth factor β1 and 5-azacytidine, and to investigate a new way to induce cells. METHODS: The rabbit abdominal fats were isolated and then the adipose-derived stromal cells were separated by col agenase digestion method for myogenetic induction with different methods: 5-azacytidine induction group, MyoD+transforming growth factor β1 induction group, 5-azacytidine+MyoD+transforming growth factor β1 combination induction group. The blank control group was set. The morphological characteristics of cells were observed at 1, 4, 8, 12, 16, 20, 24 and 28 days after induction, and the col agen type Ⅲ level were detected with 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: Compared with the other groups, the cel activity was higher in the combination induction group and reached peak at 16 days after induction, the number and volume of myotubes were increased at 20 days with regular arrangement, and desmin showed strong expression. Cel cycle showed the ratio of adipose-derived stromal cells in the DNA replication phase was increased in the combination induction group, the ratio of cells in the clearance period was decreased; the level of col agen type Ⅲ was increased significantly, and the difference was significant. The results indicate that 5-azacytidine combined with multiple factors can promote the differentiation of adipose-derived stromal cells into myoblasts with significant cel proliferation, which is considered as the ideal method to in vitro induction of myoblasts.