[Abstract] Objective To investigate the effect of exosome in cultured in vitro H9C2 myocardial cells injury of diabetic mice and its mechanism. Methods The mouse model of diabetic myocardial injury was established by using db/db mice (n=10) and their mate mice db/+ (n=5). Serum exosomes were isolated and quantitated using the exosome isolation reagent and EXOCET Quantitation kit. The serum exosomes were labeled with PKH26 (red fluorescent cell linker) to detect the endocytosis in H9C2 cells. The expressions of exocrine associated protein and inflammatory cytokines in H9C2 cells with or without exosome stimulation were detected by Western blotting. TUNEL was used to detect apoptosis. A neutralizing antibody of Rab1a was used for blocking experiment. Results Db/db mice produced more exosomes than db/+ mice (30.95×109/ml vs. 10.45×109/ml, P<0.01). Moreover, H9C2 cells cultured in vitro could swallow more serum-exosomes derived from db/db mice. Meanwhile, serum exosome from db/ db mice, as used to interfere H9C2 cells, significantly increased the expression of inflammatory cytokines, such as 6.2 folds to IL-6 and 2.6 folds to IL-1β (P<0.01). Furthermore, the apoptosis in H9C2 cells increased compared to those from db/+ mice. Mechanism studies announced that the increased expression of Rab1a in exosomes-derived from db/db mice, and blocking the expression of Rab1a in exocrine of db/db mice with Rab1a neutralizing antibody could significantly inhibit the endocytosis and apoptosis of H9C2 cells. Conclusions Serum exosomes isolated from db/db mice may trigger inflammation and apoptosis of cardiomyocytes cultured in vitro, which may be involved in the evolution of diabetic myocardial injury. Inhibition of exosome secretion or intervention of its regulatory molecules may become a new research target for the treatment of diabetic myocardial injury.

Objective:To observe the clinical efficacy of Ba-pulling and Qian-traction manipulation with neck suspension and movement for acute cervical radiculopathy. Methods: A total of 85 patients who met the inclusion criteria were randomized into an observation group and a control group by random numbers, with 43 cases in the observation group and 42 cases in the control group. The observation group was treated with Ba-pulling and Qian-traction manipulation with neck suspension and movement;while the control group was treated with Bashen-pulling and stretching manipulation in a supine position. The treatment was performed once a day, 10 times as a treatment course. The therapeutic efficacy was evaluated after 1 treatment course, and the changes in the scores of visual analog scale (VAS) and neck disability index (NDI) were observed. Results: The total effective rate was 97.7% in the observation group, and 83.3% in the control group, and the difference between the two groups was statistically significant (P＜0.05). After treatment, the VAS and NDI scores of both groups were significantly decreased (both P＜0.01), and the differences in the VAS and NDI scores between the two groups were statistically significant (both P＜0.01). Conclusion: Both Ba-pulling and Qian-traction manipulation with neck suspension and movement and Bashen-pulling and stretching manipulation in a supine position can relieve pain and improve cervical function in patients with acute cervical radiculopathy, and Ba-pulling and Qian-traction manipulation with neck suspension and movement can produce more significant efficacy than Bashen-pulling and stretching manipulation in a supine position.

OBJECTIVETo assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA).

METHODSTotally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror.

RESULTSCompared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05).

CONCLUSIONSTMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Celecoxib ; Drug Therapy, Combination ; methods ; Humans ; Knee Joint ; Osteoarthritis, Knee ; drug therapy ; Pain ; Pain Measurement ; Tendons ; Treatment Outcome

OBJECTIVETo explore early clinical effect of acetabular cup in total hip replacement for the treatment of Crowe II developmental dysplasia of hip.

METHODSEighteen patients (18 hips) with Crowe type II developmental dysplasia of hip were treated with total hip replacement from September 2001 to July 2013. Among them,including 13 males and 5 females aged from 42 to 60 years old with an average of 47.6 years old; the courses of diseases ranged from 9 to 22 years with an average of 13.5 years. All the patients had hip joint pain, limb shortening and limited hip function before operation. Harris score of hip joint were used to evaluate recovery of function at 1 day and 12 months after operation. Prosthetic coverage of acetabular cup at 1 week after operation was observed by using radiography.

RESULTSEighteen patients (18 hips) were followed up from 12 to 24 months with an average 17 months. All incisions were healed at stage I. No deep vein thrombosis, hip dislocation, periprosthetic joint infection and prosthesis loosening were occurred. No revision surgery during follow-up period. Prosthetic coverage of acetabular cup was more than 80% at 1 week after operation. Harris score were increased from 42.67 ± 5.06 before operation to 94.79 ± 3.27 at 12 months after operation (t = -45.269, P < 0.001).

CONCLUSIONFor type Crowe II developmental dysplasia of hip patients, 15° face-changing acetabular cups in THR could obtain higher actebular component coverage rate and satisfactory early clinical effects.

Acetabulum ; surgery ; Adult ; Arthroplasty, Replacement, Hip ; instrumentation ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Hip Joint ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged

OBJECTIVETo discuss the clinical effect of closed reduction and interlocking intramedullary nailing in the treatment of femoral shaft fracture.

METHODSFrom March 2006 to December 2011,103 patients with femoral shaft fracture were treated by closed reduction and interlocking intramedullary nailing including 76 males and 27 females with an average age of 36 years old ranging from 19 to 55 years old. According to AO classification,there were 64 cases with type A,27 with type B, 12 with type C. Thirteen cases were open fractures including 5 cases with Gustilo type I , 8 with Gustilo type II . The time of bone healing were observed after operation, the knee function recovery was evaluated by HSS scoring standard at 1 year after operation.

RESULTSThe intraoperative complications included femoral neck fracture in 1 case and proximal femoral fracture in 1 case,both of the patients were treated with reconstructive intramedullary interlocking nail and the fractures healed postoperatively. One patient was suffered from common peroneal nerve injury,which were fully recovered at 4 months later after medical treatment. All the patients were followed up from 12 to 28 months (averaged 22 months). All of the fractures were healed well and the average healing time was 3 to 9 months (averaged 5 months). All the hip joints were recovered to normal function. The average HSS was 90.89±5.06 at 1 year after operation.

CONCLUSIONInterlocking intramedullary nailing is the preferred treatment for patients with femoral shaft fracture. Carefully operating and closed reduction can reduce the complications.

Adult ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; instrumentation ; Fractures, Open ; surgery ; Humans ; Male ; Middle Aged ; Young Adult

Objective:Study on proliferation of gastric cancer cells (GCCs) and expressions of VEGF under the CO2 pneumoperitoneum environment,and effect of Bcl-2 specific inhibitor on the expression of VEGF in GCCs/TAMs co-culture system.Methods:TAMs induced by PMA and IL-4 in vitro.TAMs and MKN-45 cells were co-cultured in Transwell chamber,in pseudo CO2 pneumoperitoneum environment.Proliferation activity of GCCs was detected by MTT assay,and ELISA method was used to detect VEGF concentration in the cell culture supernatant.Results:Co-culture system was divided into CO2 pneumoperitoneum group and control group.Proliferation activity of MKN-45 cell and expression of VEGF of 5mmHg,10mmHg and 15mmHg groups are no difference with the control group;25 mmHg group is opposite to the former.The cells in co-culture group and CO2 pneumoperitoneum group (15 mmHg) were added to ABT-737,VEGF expression of co-culture + ABT-737 group was significantly lower than that of in the control group(P=0.001).Co-cultured cells VEGF expression in pneumoperitoneum group was significantly lower than the control group without inhibitor (P=0.000).Conclusion:Normal laparoscopic pneumoperitoneum is no stimulation of tumor cell proliferation defects,And higher pneumoperitoneum pressure might increase the hypoxia status of tumor microenvironment,promote tumor malignant progression.The interaction between tumor cells and TAMs may be achieved by paracrine Bcl-2-VEGF loop.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

Objective To explore the effect of nuclear factor-kappaB(NF-?B) on cardiac myocyte apoptosis and understand the molecular mechanism of decrease of heart function in septic shock rats.Methods Twenty Wistar rats were randomly divided into the septic shock group and the normal control group.The septic shock group(n=10) were fixed with anaesthesia and made into 1.5 cm slices just along the abdomen midline.Then the roots of appendices were returned to the belly cavity after being ligated and punctured 4 times with the No.14 pinhead.After that,the slices were sewn up layer by layer.After 12 hours,the rats were all sacrificed,the blood taken and the serum separated.In the end,the heart specimens of the septic shock group were set aside.Meanwhile,the normal control group were dealt with in the same way except that they were not subjected to cecal ligation and puncture.Then NF-?B protein levels in cardiac tissue and the index of cardiac myocyte apoptosis were measured.Results NF-?B protein levels in the septic shock group(9 cases were strong masculine gender,1 case was middle masculine gender)elevated significantly compared with the normal control group(8 cases were negative gender,2 cases were weak masculine gender) in cardiac tissue(P

Background Researches demonstrated that corneal dystrophy is associated with the mutation of transforming growth factor beta induced gene(TGFBI)located at chromosome 5q31 domine.Recent study showed that the gene mutation location is in R124H of TGFBI gene. Objective This study was to identify the mutation characteristics of TGFBI gene in a Chinese family with Avellino corneal dystrophy. Methods This Chinese family with Avellino corneal dystrophy were determined and surveyed in Peking University Third Hospital.Periphery blood from 8 patients with Avellino corneal dystrophy and 2 unaffected subjects were collected from a Chinese family with corneal dystrophy for the extraction of DNA.Exons 4,11,12 of the TGFBI gene were amplified by polymerase chain reaction(PCR),and the amplified products were sequenced directly and compared the gene sequence with that of TGFBI in GenBank.Written informed consent was obtained from each Subject prior to any medieal process. Results This family included 27 members of consecutive 4 generation.The hereditary pattern W88 in accordance with the autosomal dominant inheritance.Directly sequencing of 8 affected members revealed a G tO A transition at codon 124 (CGC to CAC),producing R124H mutation of TGFBI gene.Two synonymous single nucleotide polymorphism(SNP)of TGFBI gene occurred in the family.including a C to T transition at eodon 472(CTC to CTT)in 8 members,and a T to C transition at codon 540(TTT>TTC)in 9 members,which wag unrelated with disease. Conclusion R124H mutation of the TGFBI gene is found in this Chinese family with Avellino corneal dystrophy.

Objective To determine the effects of health education on medication compliance and bone mineral density of patients with osteoporosis in community. Methods A total of 123 patients with osteoporosis in A and B community(64 patients in control group and 59 patients in intervention group)received diverse health education and guidance. After a 2-year intervention, knowledge of osteoporosis, health behavior, medication compliance and bone mineral density change were compared between the two groups. Results In the intervention group, the proportions of correctly responding to the questions of osteoporosis were significantly improved before and after intervention(P < 0.05), except the questions of"exercise to what extent is better". Similarly, health behavior and medication compliance were also significantly improved in the intervention group(P < 0.05). In addition, knowledge of osteoporosis, most health behavior and medication compliance were better than those in the control group(P < 0.05). The difference in the bone mineral densitybetween the intervention group and the control group before and after intervention was 0.025±0.322 and -0.139±0.312(P < 0.05), respectively, showing significant differences(P < 0.05). Conclusion Health education for patients with osteoporosis may significantly improve health behavior, medication compliance, bone mineral density, and prognosis of osteoporosis.