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Objective:To evaluate the efficacy of percutaneous vertebroplasty (PVP) by percutaneous bi-level bilateral puncture in the treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fracture (ASOTLF) with dense bone bands.Methods:From March 2017 to March 2018, 65 patients (65 vertebrae) with type ⅡA ASOTLF with dense bone bands were treated at Department of Orthopaedic Trauma, Qingdao Central Hospital. They were 25 males and 40 females, with an age of (71.6±8.4) years. The time from injury to operation was (3.5±0.7) d. They were divided into 2 groups according to different treatments. In the observation group of 31 cases (31 vertebrae), PVP was conducted by percutaneous bi-level bilateral puncture; in the control group of 34 cases (34 vertebrae), PVP was conducted by percutaneous single-level bilateral puncture. The 2 groups were compared in terms of operation time, volume of polymethyl methacrylate injected, incidence of bone cement leakage, incidence of vertebral refracture, diffusive distribution of bone cement in the vertebral body, and visual analogue scale (VAS) and Oswestry dysfunction index (ODI) at postoperative 1 day and 6 months.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). Operations were completed uneventfully in all the 65 patients. All the 65 patients were followed up for (8.2±1.2) months. There was no significant difference between the 2 groups in operation time, incidence of bone cement leakage or incidence of vertebral refracture for each vertebral body injured ( P>0.05). The volume of polymethyl methacrylate injected [(6.64±0.93) mL] and the excellent and good rate of diffusive distribution of bone cement in the vertebral body [87.1% (27/31)] in the observation group were significantly larger than those in the control group [(4.36±0.79) mL and 64.7% (22/34)] ( P<0.05). The VAS scores at postoperative 1 day and 6 months [2 (1, 2) and 1 (1, 2)] and ODIs at postoperative 1 day and 6 months (23.7%±1.6% and 18.8%±1.4%) in the observation group were significantly lower than those in the control group [2 (2, 3) and 2 (2, 2); 26.9%±4.2% and 22.1%±3.3%] ( P<0.05). The VAS scores and ODIs at postoperative 1 day and 6 months in all patients were significantly lower than those before operation ( P<0.05). Compression symptoms of the spinal cord and nerve root were observed in none of the patients. Conclusion:In the treatment of type ⅡA ASOTLF with dense bone bands, PVP by percutaneous bi-level bilateral puncture is more effective than the traditional PVP by percutaneous single-level bilateral puncture, and will not increase bone cement leakage or vertebral refracture.
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Objective:To evaluate the accuracy of the domestic "Skywalker" surgical robot in implementing personalized lower limb alignment reconstruction scheme in total knee arthroplasty (TKA) and the short-term clinical outcome of robotic assisted TKA.Methods:From September 2020 to January 2021, the data of patients who received surgical robot assisted TKA in 5 clinical centers in China (Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Southwest Hospital affiliated to Third Military Medical University, The Affiliated Hospital of Qingdao University, The First Affiliated Hospital of Chongqing Medical University and Yantai Yuhuangding Hospital, and Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine was the group leader) were prospectively collected and retrospectively analyzed. There were 24 males and 82 females with an average age of 67.6±7.3 years, (range 45-80 years); Average body mass index 26.42±4.31 kg/m 2, all the operation were performed by "Skywalker" surgical robot system according to preoperative design based on CT. The operation time, intraoperative blood loss, hospitalization days and postoperative complications were recorded, and the imaging indexes including hip-knee-ankle (HKA), lateral distal angle of femur (LDFA) and medial proximal angle of tibia (MPTA) measured before and after the operation, implant model indexes (preoperative planning implant model and postoperative implant model) and short-term clinical efficacy indexes [Western Ontario and McMaster Universities (WOMAC) osteoarthritis index] pain score, stiffness score, joint function score, total score and SF-12 score before and 3 months after the operation) were compared. Results:The average follow-up period was 109.60±9.80 d, (range 95-143 d). The average operation time of 106 patients was 105.30±23.22 min; The average intraoperative blood loss was 141.70±58.33 ml; The average length of hospitalization was 5.82±2.80 d. One patient had ischemic stroke after operation, and one patient had abnormal liver function after operation. According to the judgment of the investigator, all of them were not related to the operation. The actual angle error is the difference between the preoperative planning angle and the postoperative measurement angle. The absolute error of 99.1% (105/106) of the HKA angle was within 3°, 90.8% (69/76) of LDFA, 98.7% (75/76) of the MPTA. In 45 patients in one center where data were available, the actual implant models used in all patients were consistent with the preoperative planning size, and there were only differences in version selection such as Asian condyle. WOMAC pain score, joint function score, total score was improved from 7.34±2.85, 25.10±9.85, 34.75±13.02 to 3.34±2.66, 14.68±9.64, 18.66±13.49 before and after operation, respectively, which were statistically significant ( P<0.001) and SF-12 physiological score and psychological score were improved form 27.24±6.42, 30.68±8.26 to 38.83±5.74, 39.36±7.85 before and after operation, respectively, which were statistically significant ( t=7.33, P<0.001; t=4.53, P=0.043). Conclusion:Domestic surgical robot system "Skywalker" can assist the surgeon to achieve accurate and personalized reconstruction of lower limb alignment and achieve satisfactory short-term clinical outcomes. The long-term clinical outcomes of personalized reconstruction and survival rate of implant still need to be further studied.
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Objective:To analyze the epidemiological characteristics and clinical manifestations of patients with acute brucellosis in Zaozhuang City of Shandong Province, and to improve the understanding of brucellosis.Methods:The demographic characteristics, epidemiological characteristics, clinical manifestations and laboratory examination results of patients with acute brucellosis diagnosed in Zaozhuang Municipal Hospital from January 2019 to December 2020 were retrospectively analyzed.Results:Most of the 71 patients with acute brucellosis were male, with a male-to-female ratio of 2.2 ∶ 1.0 (49 ∶ 22) and an average age of (53.23 ± 14.12) years old, mainly in Yicheng District (32.39%, 23/71); 80.28% (57/71) of patients had a history of epidemiological contact; the disease occurred throughout the year, mainly from April to October (76.06%, 54/71); the main symptoms were fever (100.00%, 71/71), muscle and joint pain (80.28%, 57/71), fatigue (54.93%, 39/71), hyperhidrosis (39.44%, 28/71), hepatosplenomegaly (18.31%, 13/71) and orchitis (4.23%, 3/71), etc. Laboratory examination results showed that decreased white blood cells accounted for 12.68% (9/71), decreased red blood cells accounted for 21.13% (15/71), thrombocytopenia accounted for 8.45% (6/71), decreased lymphocyte percentage accounted for 12.68% (9/71), increased C-reactive protein accounted for 67.61% (48/71), and accelerated erythrocyte sedimentation rate accounted for 50.70% (36/71). Some patients were accompanied by elevated alanine aminotransferase (50.70%, 36/71) and aspartate aminotransferase (42.25%, 30/71), 57 patients (80.28%) with positive blood culture.Conclusions:The clinical manifestations of acute brucellosis are atypical in Zaozhuang City. Brucellosis screening should be carried out for patients with long-term unexplained fever and an epidemiological exposure history, especially those with muscle joint pain, hepatosplenomegaly and orchitis.
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Objective:To analyze the rules and distribution of acupuncture points used in pediatric tuina treatment for autumn diarrhea, and to provide a theoretical basis for the promotion and application of Tuina treatment techniques for pediatric diarrhea.Methods:We searched the literature on Tuina treatment of pediatric autumn diarrhea in the Chinese academic journals full-text database (CNKI), Chinese science and technology journals full-text database (VIP), Wanfang database and Chinese biomedical database (SinoMed), and collected the Tuina prescriptions on children's autumn diarrhea disease from the database to March 2021,with the help of the Traditional Chinese Medicine Inheritance Support Platform (V 2.0) software,and Data mining technology was applied to analyze the common acupoints and acupoint distribution of pediatric Tuina treatment of autumn diarrhea to form the core acupoints used in pediatric Tuina treatment of autumn diarrhea.Results:A total of 101 pediatric autumn diarrhea Tuina prescriptions were screened, involving 61 acupoints, among them 18 acupoints used more frequently, and 35 acupoint combinations were obtained, including 15 two-acupoint combinations, 11 three-acupoint combinations, and 2 four-acupoint combinations, and 7 new acupoint combinations were obtained.Conclusion:The core points mainly belong to the spleen meridian, large intestine meridian, abdomen, seven knotted bones, turtle tail, spine, and umbilicus, which can be used as a basic Tuina prescription for clinical reference.
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Objective:To evaluate the diagnostic values of serum fibrinogen and D-dimer for periprosthetic joint infection (PJI).Methods:The medical records were retrospectively analyzed of the 175 patients who had undergone hip or knee revisions at Department of Joint Surgery, Affiliated Hospital to Qingdao University from August 2013 to June 2019. Of them, 59 were diagnosed as PJI (31 knees and 28 hips), including 33 males and 26 females with an age of (67.4±11.7) years and a body mass index (BMI) of (26.1±3.6) kg/m 2 while 116 as aseptic loosening (AL) (19 knees and 97 hips), including 67 males and 49 females with an age of (70.3±8.9) years and a BMI of (25.0±3.6) kg/m 2. The plasma levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), fibrinogen and D-dimer in the 2 groups were recorded and analyzed. The receiver operating characteristic curve (ROC) was used to calculate the sensitivity and specificity of each indicator; the diagnostic value for each indicator was calculated according to the area under the curve (AUC). Results:There was no significant difference between the PJI and AL groups in gender, age or BMI ( P>0.05), but there was a significant difference in the joint type ( P<0.05). Compared with the AL group, the PJI group had significantly higher levels of CRP, ESR, fibrinogen and D-dimer ( P<0.05). The AUCs for CRP, ESR, fibrinogen and D-dimer were 0.830, 0.850, 0.848 and 0.664, respectively. By the Youden index, the optimal predictive cutoffs for CRP, ESR, fibrinogen and D-dimer were 8.06 mg/L, 17.60 mm/h, 3.73 g/L and 685.00 ng/mL, giving sensitivities of 79.2%, 85.4%, 81.3% and 64.6% and specificities of 85.7%, 76.2%, 79.8% and 61.9%. Conclusions:The diagnostic value of serum fibrinogen may be high for PJI, similar to that of CRP or ESR. However, D-dimer may be of limited value for diagnosis of PJI.
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Objective:To evaluate the curative effect of 3D printing titanium trabecular metal （TTM） acetabular cups in revision of total hip arthroplasty （THA）.Methods:A retrospective case series study was conducted on the clinical data of 24 patients （25 hips） undergoing THA revision in Affiliated Hospital of Qingdao University from May 2016 to September 2019. There were 10 males and 14 females，aged 43-78 years ［（63.0 ± 11.4） years］. According to Paprosky classification，5 patients（5 hips） were classified as type Ⅰ，9 patients （10 hips） as type ⅡA，5 patients （5 hips） as type ⅡB，1 patient （1 hip） as type ⅡC and 4 patients （4 hips） as type ⅢC. All patients used 3D printing TTM cups. Seven patients （7 hips） were combined with 3D printing TTM pads，and 3 patients （3 hips） were combined with structural bone grafts. The visual analogue scale （VAS），Harris score and quality of life Health Survey Scale （SF-36） score were evaluated before operation，3 months after operation，12 months after operation and at the last follow-up. The X-ray film of hip joint was performed to evaluate the upward movement distance of hip center of rotation （HCOR），limb-length discrepancy （LLD） before operation，1 day after operation and at the last follow-up. The position of cups and bone ingrowth were evaluated at the last follow-up. The postoperative complications were observed.Results:All patients were followed up for 14-54 months ［（34.2 ± 9.3）months］. The VAS was （4.0 ± 0.7）points，（2.3 ± 0.8）points and （0.9 ± 0.2）points at postoperative 3 months，12 months and the last follow-up，significantly lower than that preoperatively ［（6.1 ± 1.0）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative VAS at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The Harris score was （64.6 ± 5.3）points，（80.5 ± 3.7）points，and （90.3 ± 3.6）points at postoperative 3 months，12 months and the last follow-up，significantly higher than that preoperatively ［（38.8 ± 6.2）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative Harris score at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The SF-36 score was （556.3 ± 21.9）points，（711.6 ± 15.9）points and （752.8 ± 23.0）points at postoperative 3 months，12 months and the last follow-up，significantly higher than that preoperatively ［（326.3 ± 30.7）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative SF-36 score at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The upward movement distance of HCOR was （13.5 ± 2.6）mm and （13.6 ± 2.6）mm on the first day after operation and at the last follow-up，significantly lower than that preoperatively ［（34.1 ± 3.5）mm］ （ P < 0.05）. The LLD was （6.2 ± 1.8）mm and （5.3 ± 1.5）mm on the first day after operation and at the last follow-up，significantly lower than that preoperatively ［（31.6 ± 5.2）mm］ （ P < 0.05）. All the cups were stable. Good bone ingrowth was found in 23 hips. There was no postoperative complication except for one patient with poor wound healing. Conclusion:For THA revision，the use of 3D printing TTM cups can effectively relieve pain，improve hip function，provide satisfactory biological fixation and and reduce complications.
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Objective To compare the perio-perative effects of modified bikini minimally invasive approach versus direct lateral approach for total hip arthroplasty(THA).Methods A total of 60 patients(70 hips),including 45 males (49 hips) and 15 females (21 hips) aged (54.9 ± 6.1) years (range from 38 to 63 years)who underwent THA for the first time at joint surgery department of our hospital were retrospectively analyzed from October 2015 to May 2017.Thirty patients(34 hips) underwent modified bikini approach THA(the modified bikini group),and 30 patients(36 hips) received direct lateral approach THA(the direct lateral group).The comparisons were performed between the two groups in the orthopaedic pelvic films,disease stages,Harris scores before and after operation,intra-operative blood loss,postoperative drainage,blood transfusion rate,24-h visual analog scale(VAS) scores for pain after operation,operation time and incision length.The straight leg elevation time,recovery time of hip abduction muscle strength,average hospitalization days and subjective satisfaction of patients were evaluated.Relevant surgical complications were recorded.Results Postoperative X-ray examinations showed that the position of prosthesis was good in both groups.There were no statistically significant difference between the two groups in operation time,intraoperative blood loss,postoperative drainage,blood transfusion rate,straight leg elevation time and subjective satisfaction(P＞0.05).There were statistically significant differences between the modified bikini group and the direct lateral group in the incision length[(9.2±1.9) cm vs.(18.5±3.3) cm],recovery time of hip abductor muscle strength [(1.6 ± 0.8) d vs.(12.1 ± 3.9) d],Harris scores [(87.1±5.9) vs.(86.8 ± 7.1)],24-hour VAS scores [(4.6 ± 2.3) vs.(7.0 ± 3.6)] and average hospitalization days[(7.4 ±1.8) vs.(10.8 ±2.4)]after operation between the modified bikini group and the direct lateral group(P＜0.05).One case with lateral femoral cutaneous nerve injury and 1 case of posterior cortical perforation of proximal femur were found in the modified bikini group,and 1 case of dislocation of hip prosthesis was observed in the direct lateral group.Conclusions Both of the modified bikini minimally invasive approach and direct lateral approach for THA can achieve better postoperative effect,while the modified bikini approach achieves faster recovery.
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Objective To evaluate the effect of TBL (team-based learning) method in pharmacology teaching for the foreign students of clinical medicine. Methods In the course of pharmacology teaching of the foreign students of clinical medicine, TBL method was performed in the 2012-year students and tradi-tional teaching method was performed in the 2011-year students. After the teaching, students' grades in the ordinary performance, their final exam scores and their evaluation of the two teaching methods were com-pared. Graph pad 5 was used to analyze the data and the t test was performed. Results The average ordi-nary performance of the students with TBL was significantly higher than that with the traditional teaching [(84.94 ±12.66) vs. (72.30 ±4.90), P=0.000] and the final examination scores were significantly im-proved [(74.00±6.76) vs. (69.00±6.20), P=0.023]. The survey showed students were more satisfied with the TBL teaching mode than traditional teaching mode [(8.40±0.71) vs. (7.12±1.07), P=0.000]. Conclusion TBL teach-ing mode can effectively improve the pharmacology teaching effect of foreign students.
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Aim To investigate the effects of DGK in-hibitor R59022 on ET-1-induced myocardial hypertro-phy and autophagy, and explore the possible mecha-nisms. Methods Myocardial hypertrophy was in-duced by ET-1 in cultured rat neonatal cardiomyo-cytes. Western blot was used to detect the expression of microtubule-associate protein 1 light chain 3 ( LC3 ) , beclin-1, p62, p-Akt and Akt. mRNA expression of brain natriuretic peptide ( BNP) and beta mysion heav-y chain (β-MHC) and the cell size of cardiomyocytes were detected by RT-PCR and immunofluorescence, respectively. Results Treatment cardiomyocytes with ET-1(10 -7 mol·L-1 ) for 24 h induced the myocardi-al hypertrophy in cultured neonatal rat cardiomyocytes with the activation of autophagy as evidenced by the in-creased expression of autophagy-related proteins LC3-II/I and beclin-1 , as well as the increased p62 degra-dation. While, myocardial hypertrophy induced by ET-1 , including the increased myocardial cell size and the mRNA expression of fetal gene BNP and β-MHC, could be reversed by autophagy inhibitor 3-methyl ade-nine (3-MA) and chloroquine ( CQ) ,but promoted by autophagy agonist rapamycin ( RAPA ) . Pretreatment cardiomyocytes with R59022, an inhibitor of DGK, en-hanced ET-1-induced myocardial hypertrophy by en-hancing autophagy in cardiomyocytes. Furthermore,ET-1 treatment inhibited the activation of Akt by the down-regulation of the Akt phosphorylation, and R59022 en-hanced the effect of ET-1 on the activation of Akt. Conclusions Enhanced autophagy contributes to car-diomyocyte hypertrophy. R59022 deteriorate ET-1-in-duced myocardial hypertrophy by activating autophagy. The possible mechanism may be related to the inhibi-tion of activation of mTOR signaling pathway by inhibi-ting the activation of Akt.
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Objective To investigate whether the new mechanically-activated (MA) cation channel Piezo 1 protein can cause the apoptosis of human chondrocytes under compressive loading,using a Flexercell unit by activating MAPK/ERK5 signal pathway.Methods Primary human chondrocytes were isolated,cultured,and then subjected to the static compressive loading for 2 h,12 h,24.h and 48 h,respectively.The GsMTx4,which is the special inhibitor of the Piezo1 protein and the BIX02188,which is the inhibitor of the ERK5 served as the inhibitor group.The immunofluorescence was used to locate the expression of the Piezo 1 protein.The expressions of Piezo1 and ERK5 were assessed by reverse transcription-polymerase chain reaction (RT-PCR),as well as the apoptosis gene B cell lymphoma/leukemia-2 (Bcl-2),Bcl-associated X protein (Bax) and Bcl-associated death promoter (BAD).In addition,Piezo1 inhibitor,GsMTx4,was used to block the MA cation channel Piezo1,served as the inhibitor group.AVPI was used to detect the apoptosis of the OA chondrocytes.Results The location of the Piezo1 was expressed in nucleus and cytoplasm of chondrocytes.The expression of the Piezo1,ERK5,BAD,and Bax mRNA in the OA chondrocytes is weak.The 12 h stretch force group was significant increased,and the 24 h stretch force group was the highest expression.However,the expression of the 48 h group was decreasing.The expression of the Bcl-2 in the 12 h group was decreasing,and the 12 h stretch force group was the lowest expression while the 24 h stretch force group was increasing.In the GsMTx4 group,the expression of the Piezo1,ERK5,BAD,Bax was decreasing while the Bcl-2 was increasing.In the BIX02188 group,the expression of the ERKS,BAD,Bax was increasing while the Bcl-2 was decreasing,while the expression of the Piezo1 was not change.The result of AV-PI shown that the 2 h stretch force group increased early stage of apoptosis.The 12 h stretch force group increased late stage of apoptosis,and the 24 h stretch force group's apoptotic rate was the highest.However,the apoptotic rate of the 48 h group was lower than the 24 h stretch force group.The GsMTx4 could inhibit the late stage of apoptosis.Conclusion Piezo 1 plays an important role in the apoptosis of human chondrocyte through the MAPK/ERK5 signal pathway.
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BACKGROUND:Excessive mechanical stimulation can lead to endoplasmic reticulum stress. Endoplasmic reticulum stress plays an important role in the occurrence and development of degenerative diseases. Caspase-12 is a specific molecule of endoplasmic reticulum stress, and the intensity of endoplasmic reticulum stress can be reflected by caspase-12 expression. OBJECTIVE:To observe the expression rule of Caspase-12 in chondrocytes under the cyclic tensile stress. METHODS:Human chondrocytes were used as test subjects. After group assignment, mechanical loading system was used. The loading group and corresponding Z-ATAD-FMK inhibitor group received 2, 12, 24, 36, and 48 hours of mechanical stimulation. The blank group (0 hour) and the paired Z-ATAD-FMK inhibitor group received the same process as the loading group except the loading. After loading, cel morphology and growth were observed under the microscope. RT-PCR and western blot assay were used to detect the expression changes of caspase-12 gene and protein in each group. RESULTS AND CONCLUSION:(1) Morphological observation results demonstrated that apoptosis appeared at 2 hours after loading, peaked at 24 hours. With prolonged time, cel growth showed the trend along stress, but apoptosis weakened. It is indicated that cyclic tensile stress could make chondrocyte apoptosis. Endoplasmic reticulum stress might be activated. The model of cel s in vitro was established successful y. (2) Caspase-12 gene and protein expression showed consistent trend, and peaked at 24 hours, which showed significant differences as compared with the blank and inhibitor groups (P<0.05). (3) Cyclic tensile stress can induce chondrocyte endoplasmic reticulum stress, and affect caspase-12 expression.
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BACKGROUND:Total knee arthroplasty had been generaly accepted as the final treatment plan, relieving pain and reconstructing function of knee joint. However, whether drainage tube can be used after replacement is stil controversial. OBJECTIVE:To compare the clinical effects of drainageversusnondrainage after primary unilateral total knee arthroplasty. METHODS:Total 102 patients undergoing primary unilateral total knee arthroplasty were randomly divided into 2 groups. In the drainagegroiup, a drainage tube was used. In the nondrainage group, drainage tube was not used. Total blood loss was calculated by recording the hemoglobin and hematocrit before operation and that after 1, 3, 7 days of operation. The pain visual analogue scale scores, arthrocele, ecchymosis, infection rate, and deep venous thrombosis of lower extremity were examined and analyzed postoperatively. Knee Society Scores were recorded at 1 year postoperatively. Above indexes were compared between the two groups. RESULTS AND CONCLUSION:(1) Total blood loss and blood transfusion rate were significantly higher in the drainage group than in the nondrainage group (P 0.05). (3) No significant difference in Knee Society Scores was detected between the two groups (P> 0.05). (4) Results indicated that the total blood loss and blood transfusion rate may decrease significantly in patients without wound drainage after total knee arthroplasty. Limb sweling and ecchymosis area were not increased. No significant difference in infection, deep venousthrombosis of lower extremity and knee function was detectable between the two groups. Thus, total knee arthroplasty without wound drainage is safe and does not have obvious adverse consequences.
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BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.
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<p><b>OBJECTIVE</b>To explore the effects of astragalus (AST) , total flavone of astragalus (TFA), total saponins of astragalus (TSA) and astragalus polysaccharides (APS) on ischemia/reperfusion (40 min/60 min) injury in isolated guinea-pig heart.</p><p><b>METHODS</b>Isolated guinea-pig hearts underwent ischemia, then followed by K-H perfusion (I/R group), AST (60 mg/L),AST (60 mg/L), TFA (60 mg/L), TSA (60 mg/L) and APS (60 mg/L) perfusion (n = 6 each).Isolated hearts without ischemia serve as control group (n = 6). Activity of lactate dehydrogenas (LDH) and creatine kinase (CK) in effluent were measured.Infarct size, myocardial superoxide dismutase (SOD) activity and malondiadehyde (MDA) contents were also determined.</p><p><b>RESULTS</b>Compared to control hearts, heart rate, coronary flow and myocardial superoxide dismutase (SOD) activity were significantly reduced, while LDH and CK in effluent as well as myocardial MDA were significantly increased in the I/R hearts during reperfusion (all P < 0.05), these changes could be partly reversed by AST and TFA perfusion.Infarct size was also significantly reduced in AST (11.9 ± 2.03) % and TFA (13.31 ± 1.17) % treated hearts compared to that in I/R group (18.9 ± 2.27) % (all P < 0.01).</p><p><b>CONCLUSIONS</b>The findings indicate that AST and TFA could attenuate I/R injury in isolated guinea-pig heart possibly through enhancing the activity of SOD and reducing lipid peroxidation.</p>
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Animals , Astragalus Plant , Guinea Pigs , Heart , Heart Rate , Myocardial Ischemia , Myocardial Reperfusion Injury , Drug Therapy , Myocardium , Plant Extracts , Pharmacology , Reperfusion Injury , Superoxide DismutaseABSTRACT
Objective To investigate the effects of aggrecanase-2 knockdown in chondrocytes from rheumatoid arthritis patient by shRNA infection.Methods Cartilage harvested from rheumatoid arthritis patients who underwent total knee arthroplasty was digested by pancreatin and type Ⅱ collagen enzyme to obtain chondrocytes.Then chondrocytes were cultured and passaged to second or third generation.After aggrecanase-2 shRNA5,aggrecanase-2 shRNA6,aggrecanase-2 shRNA7,aggrecanase-2 shRNA8 infection,growth and morphological changes of the chondrocytes were examined.To select the best target sequence,mRNA expression of aggrecanase-2 and aggrecan was detected by RT-qPCR assay on day 2,5,10,represented with Ct (Cycle threshold) value.Expression of aggrecan protein was detected by immunocytochemistry,represented with IOD (integral optical density) value.Results aggrecanase-2 knockdown had no obvious effects on the morphology and growth of the chondrocytes.MOI (multiplicity of infection) was 100,50,25,and infection efficiency was 90％,60％,30％ with the corresponding viral load of 200 μl,100 μl,50 μl after transfection.mRNA expression of aggrecanase-2 was suppressed significantly,especially the group of shRNA5 which suppressed aggrecanase-2 expression from 0.876 3±0.115 6 to 0.069 9±0.015 1 (P ＜ 0.05).mRNA and protein expression of aggrecan were significantly upregulated after infection.mRNA expression of aggrecan increased from 0.992 1±0.201 3 to 3.049 2±0.278 2 (P ＜ 0.05) and protein expression of aggrecan increased from 496.160 5± 225.673 7 to 4 525.433 0±1 131.813 0 (P ＜ 0.05).Conclusion aggrecanase-2 suppression in chondrocytes by lentivirius infection is an effective method to protect the expression of aggrecan.
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Objective To establish a model of full-thickness avascular meniscal defect to assess outcome of bone-marrow mesenchymal stem cells (BMSCs) modified with human insulin-like growth factor Ⅰ (hIGF-Ⅰ) gene and compounded with injectable calcium alginate gel in repair of meniscal defect.Methods Models of full-thickness defect were created in the anterior comer of meniscus in goats,an area lacking of blood supply.The trial categorized the models to four groups:gene-ehanced tissue engineering (GETE) group (hIGF-Ⅰ transfected BMSCs were mixed with calcium alginate gel),BMSCs group (BMSCs were mixed with calcium alginate gel),empty group (calcium alginate gel was used alone) and control group (the defect was excluded from repair).Macroscopy was done at 4,8,and 16 weeks after operation.Variation of repair tissue was observed by light and scanning electric microscopy and aggrecan in repair tissue was determined as well.Results Meniscal defect was on the mend at 4-16 weeks after operation in GETE group,with the defect area being thoroughly filled with the white,elastic and tight repair tissue similar to normal meniscal tissue.Macroscopic examination showed a better result in GETE group than that in other groups.Light microscopy showed that repair tissue which was mainly fibrochondrocytes was arranged in line with calcium alginate fibers and that space between the fibers was mostly crammed with the matrix secreted by those cells.At the same time,those cells were tightly arranged and the matrix secreted by those cells was equally distributed according to light microscopy.Electroscopy demonstrated neat and tight arrangement of fibers and tight extracellular matrix in fiber space in GETE group.Aggrecan concentration in GETE group was relatively higher than in other groups,but still had difference from the normal meniscus.Conclusion hIGF-Ⅰ gene-transfected BMSCs combined with injectable calcium alginate gel can improve the effect in repair of full-thickness meniscal defect.
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BACKGROUND: Bone morphogenetic protein (BMP) and its derived scaffold or vector here been widely used in experiment and gradually applied in the clinic. However, the tracing method challenges the therapeutic effect, The discovery and application of enhanced green fluorescent protein (EGFP) can solve this problem.OBJECTIVE: To construct the bicistronic eukaryotic vector pIRES2-EGFP-hBMP-2 and to observe its expression in human embryo kidney (HEK) 293 cells.METHODS: hMBP-2 gene was extracted from human osteosarcoma by RT-PCR and inserted into PMD18-T vector. Following the DNA sequence verification, it was then sub-cloned into the eukaryotic vector pIRES2-EGFP. After restriction enzyme analysis the pIRES2-EGFP-BMP-2 was transfected into HEK 293 cells. Then its expression was observed using fluorescence microscopy and Western blot.RESULTS AND CONCLUSION: hBMP-2 gene was amplified and the eukaryotic vector plRES2-EGFP-hBMP-2 was constructed successfully. At 48 hours after tranfection of pIRES2-EGFP-hBMP-2 into HEK 239 cells, approximately 30% of cells emitted green fluorescence under a fluorescence microscope. Western blot results demonstrated that there was a specific BMP strap at Mr46×10s side, which indicated that the target gene could be expressed in constructed pIRES2-EGFP-hBMP-2 vector.
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BACKGROUND: The existed repair method for cartilage defects has shortcomings of insufficient repairing tissue numbers, poor biomechanical properties, as well as donor site complication. Thus it is deficient to repair large-sized osteochondral defects using one method.OBJECTIVE: To investigate the therapeutic effect of tissue engineering modified Mosaicplasty on repairing large-sized osteochondral defects.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Center Laboratory of Qingdao University Medical College from January to September 2009.MATERIALS: The hircine bone mesenchymal stem cells (BMSCs) were in vitro cultured, and resuspended with algin solution to obtain BMSCs-calcium alginate gel.METHODS: Totally 12 goats were prepared for osteochondral defects models and were divided into 3 groups.BMSCs-Mosaicplasty group, BMSCs compound with injectable alginate calcium gel was then applied to fill the "dead space" after Mosaicplasty. In the Mosaicplasty group, the defects were repaired by Mosaicplasty. There was no treatment in the control group.From 4 to 16 weeks postoperatively, the animals were sacrificed and the in gross and under electromicroscopy.MAIN OUTCOME MEASURES: ①Gross observation: the joint was exposed to observe the repair effect at weeks 4, 8,16 after operation. ②Histological examination: specimens were harvested at 16 weeks after operation and observed by haematoxylin-eosin staining, toluidine blue staining under light microscopy. ③Transmission electron microscope was used at 16 weeks after operation.RESULTS: The transplanted subchondral bone and superficial cartilage was integrated hardly with each other or with recipient sites in tissue engineering modified Mosaicplasty groups at 16 weeks after operation. The quality and appearance of the transplanted and regenerated cartilage was similar to normal hyaline cartilage. Under microscopy, the regenerated cartilage was integrated with neighbor tightly in regular arrange. ECM distributed evenly and deeply stained by alcian blue. There was no obviously repaired in the control group.CONCLUSION: Tissue engineering can ameliorate the outcome of Mosaicplasty to repair the osteochondral defects.
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[Objective]To discuss the results of the GIEBEL system in high tibial osteotomy to treat varus knee.[Method]Nineteen patients with medium varus knee were selected to be treated with osteotomy.Preoperatively,the mean varus angle was 12.29?,without deformity in other department of the knee joint.The main symptom was the pain in medial department.Accurate high tibial osteotomy with GIEBEL system fixation was performed in 31 knees of these patients.Weight-bearing extremity X-ray testing,varus angle,mechanical axle and anatomical axle were marked preoperatively and postoperatively.Joint space discrepancy between lateral and medial joint,the range of motion,Lysholm assessment and objective satisfactory survey were also performed to determine the outcome of the treatment.[Result]The mean follow-up of nineteen patients was 22 months.During this period,one case of temporary common fibular nerve paralysis occurred and healed at several days.All of the osteomy interface achieved bony healing.The corrected abnormality was 12.32?.The mechanism axle of the extremity maintained as the level just after operation during the follow-up and no degeneration of the joint space aggravated.The excellent and good rate of the treatment was 89.5%,and 84.2% of the patients expressed satisfactory for the operation.No difference of the range of motion was found before and after operation,but the Lysholm score,joint space discrepancy between lateral and medial joint and varus angle were improved after the osteotomy.[Conclusion]The GIEBEL system can fixate the bony interface of the high tibial osteotomy effectively,and possess the advantages of minimal invasion,stronger intensity and higher healing rate.
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0.05).SK & F96365 at the concentration of 5～20 ?mol?L-1 inhibited the Ca2+ influx induced by 1.0 ?mol?L-1 Thapsigargin in a concentration-dependent manner.The inhibitory effect of SK & F96365 on Ca2+ influx was decreased by overexpression of ClC-3 protein.Conclusion ClC-3 chloride channel was involved in the regulation of store-operated Ca2+ entry(SOCE).