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Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.
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Objective:To investigate the clinical efficacy of carbomer hemorrhoid gel in the treatment of second-degree internal hemorrhoid bleeding.Methods:A total of 160 anorectal outpatients with second-degree internal hemorrhoid bleeding who received treatment in Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine from January 2017 to January 2021 were included in this study. They were randomly divided into an observation group and a control group ( n = 80/group). In the observation group, carbomer hemorrhoid gel was used to plug the anus, while in the control group, a hemorrhoid suppository was used to plug the anus. All patients were treated for 7 days. Clinical efficacy was compared between the two groups. Results:After 4 days of treatment, the bleeding score in the observation group was lower than that in the control group [1(0) point vs. 2(1) points, Z = -6.70, P < 0.05). After 7 days of treatment, the bleeding score in the observation group was significantly lower than that in the control group [0(1) point vs. 1(1) point, Z = -4.73, P < 0.05]. After 4 days of treatment, there was no significant difference in the size score of the hemorrhoids between the two groups ( P > 0.05). After 4 days of treatment, the size score of hemorrhoids in the control group did not differ significantly compared with before treatment ( P > 0.05). After 4 days of treatment, the size score of hemorrhoids in the observation group differed significantly compared with before treatment ( Z = -3.16, P < 0.05). After 7 days of treatment, the size score of the hemorrhoids in the observation group was lower than that in the control group [1(1) point vs. 1(0) point, Z = -4.48, P < 0.05]. The total response rate in the observation group was significantly higher than that in the control group [97.5% (78/80) vs. 75% (60/80), Z = -4.50, P < 0.05]. Conclusion:Carbomer hemorrhoid gel is a new drug used to treat hemorrhoids. It has a new dosage form, has no stimulation to the rectum, and is safe to use. Carbomer hemorrhoid gel is highly effective on second-degree internal hemorrhoid bleeding and deserves clinical popularization.
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BACKGROUND: The Id2 gene is an endogenous negative regulator of basic helix-loop-helix factor, which is involved in the cell proliferation, differentiation and existence. Id2 also shows functional diversity in the progression and infiltration of different types of tumors OBJECTIVE: To observe the changes of proliferation and invasiveness of PC-3 human prostate cancer stem cells after shRNA-Id2 transfection. METHODS: PC-3 human prostate cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of CD44+CD24-on the surface of tumor stem cells was detected by flow cytometry. The shRNA-Id2 expression vector was constructed and transfected into PC-3 human prostate cancer stem cells. Untransfected PC-3 human prostate cancer stem cells were used as control. At 48 hours after transfection, the expression of Id2 gene and protein in shRNA-Id2 transfected prostate cancer stem cells, NC-shRNA empty vector transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by RT-PCR and western blot, respectively. The proliferation and invasion of shRNA-Id2 transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by MTT assay and Transwell chamber, respectively. The expressions of E-cadherin, vimentin and Twist were detected by western blot and RT-PCR. RESULTS AND CONCLUSION: Tumor stem cell spheres were successfully isolated by the serum-free suspension culture. The expression rate of CD44+CD24-on the surface of the third-generation PC-3 human prostate cancer stem cells was (85.69±8.96) ％, indicating that the cultured tumor stem cell spheres overexpressed the phenotype of tumor stem cells. At 48 hours after transfection, the expression of Id2 gene and protein was significantly lower in the shRNA-Id2 transfection group than the non-transfection group (P < 0.05) , indicating that the expression of Id2 was successfully interfered with the expression of PC-3 prostate cancer stem cells. The invasive ability of the cells in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). Western blot and RT-PCR detection showed that the expression of E-cadherin, an epithelial marker of PC-3 prostate cancer stem cells, in the shRNA-Id2 transfection group was significantly higher than that in non-transfection group (P < 0.05) , while the expression of vimentin, a marker of mesenchymal stem cells, and Twist, a transcription factor regulating cell-mesenchymal transformation, in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). These findings indicate that RNA interference with Id2 gene can inhibit the proliferation and invasion of PC-3 prostate cancer stem cells by regulating the expression of E-cadherin, vimentin and Twist.
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BACKGROUND: The Id2 gene is an endogenous negative regulator of basic helix-loop-helix factor, which is involved in the cell proliferation, differentiation and existence. Id2 also shows functional diversity in the progression and infiltration of different types of tumors OBJECTIVE: To observe the changes of proliferation and invasiveness of PC-3 human prostate cancer stem cells after shRNA-Id2 transfection. METHODS: PC-3 human prostate cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of CD44+CD24-on the surface of tumor stem cells was detected by flow cytometry. The shRNA-Id2 expression vector was constructed and transfected into PC-3 human prostate cancer stem cells. Untransfected PC-3 human prostate cancer stem cells were used as control. At 48 hours after transfection, the expression of Id2 gene and protein in shRNA-Id2 transfected prostate cancer stem cells, NC-shRNA empty vector transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by RT-PCR and western blot, respectively. The proliferation and invasion of shRNA-Id2 transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by MTT assay and Transwell chamber, respectively. The expressions of E-cadherin, vimentin and Twist were detected by western blot and RT-PCR. RESULTS AND CONCLUSION: Tumor stem cell spheres were successfully isolated by the serum-free suspension culture. The expression rate of CD44+CD24-on the surface of the third-generation PC-3 human prostate cancer stem cells was (85.69±8.96) ％, indicating that the cultured tumor stem cell spheres overexpressed the phenotype of tumor stem cells. At 48 hours after transfection, the expression of Id2 gene and protein was significantly lower in the shRNA-Id2 transfection group than the non-transfection group (P < 0.05), indicating that the expression of Id2 was successfully interfered with the expression of PC-3 prostate cancer stem cells. The invasive ability of the cells in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). Western blot and RT-PCR detection showed that the expression of E-cadherin, an epithelial marker of PC-3 prostate cancer stem cells, in the shRNA-Id2 transfection group was significantly higher than that in non-transfection group (P < 0.05), while the expression of vimentin, a marker of mesenchymal stem cells, and Twist, a transcription factor regulating cell-mesenchymal transformation, in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). These findings indicate that RNA interference with Id2 gene can inhibit the proliferation and invasion of PC-3 prostate cancer stem cells by regulating the expression of E-cadherin, vimentin and Twist.
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Objective To discuss the clinical value of magnetic resonance diffusion-weighted imaging (MR-DWI) in distinguishing tumor remnants from tumor necrosis of pancreatic carcinoma after cryoablation treatment.Methods Conventional MRI T1WI,T2WI scan,DWI sequence and dynamic enhanced MRI scan were performed in 26 patients with pancreatic carcinoma who were received cryoablation treatment.The changes in MRI signals after cryoablation treatment were recorded.The apparent diffusion coefficient (ADC) values of the normal pancreas,preoperative tumor tissue,postoperative remnants and necrosis tissue were calculated,and the results were compared.The correlation between the ADC values and the size of the tumor was evaluated,and the differences in ADC values among the tumors that had different diameter,location and staging were statistically analyzed.Results Of the 26 patients,complete necrosis of tumor was obtained in 16.The necrotic tumor tissue displayed low-signal on T1WI,high-signal on T2WI and low-signal on DWI,with no enhancement on dynamic enhanced imaging.Active residual tumor tissue was detected in 9 patients,among them the residual tumor diameter ＞5 cm was seen in 7 patients;the residual rate was 34.6％.ADC values of the following tissue,from low to high in order,were preoperative pancreatic tumor tissue (1.022± 0.126)x10-3 mm2/s,postoperative residual tumor tissue (1.130±0.155)x10-3 mm2/s,normal pancreatic tissue (1.924±-0.124)×10-3 mm2/s and postoperative necrosis tissue (2.312-±0.214)×10-3 mm2/s.No statistically significant difference in ADC values existed between preoperative pancreatic tumor tissue and postoperative residual tumor tissue (P=0.452),while statistically significant difference in ADC values existed between normal pancreatic tissue and postoperative necrosis tissue (P＜0.001).The ADC values of pancreatic tumor tissue bore a negative correlation with the tumor size (R=-0.43,P=0.027 2),while the ADC values lacked the relationship to the tumor location as well as to the tumor staging (P=0.738 8 and P=0.089 5 respectively).Conclusion MR-DWI can effectively distinguish the residual tumor tissue from the necrotic tumor tissue of pancreatic carcinoma after cryoablation treatment,which provides reliable basis for further clinical diagnosis and treatment.
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BACKGROUND:Animal models of osteoporosis play an important role in the research of the pathogenesis, occurrence and development of osteoporosis, as wel as in the clinical diagnosis, prevention and treatment of osteoporosis. OBJECTIVE: To summarize and discuss the establishment and research ideas of osteoporosis models, explore the current situation and advance of osteoporosis models, compare the advantages and disadvantages of various methods, and provide evidence for clinical investigation. METHODS: A computer-based online search was conducted in SinoMed, VIP, Wanfang and PubMed databases by using the key words of “animal model, osteoporosis” from January 1969 to October 2015. The language was limited to both Chinese and English. Relevant articles were screened according to inclusion and exclusion criteria. The documents about the methods of osteoporosis model preparation, method improvement as wel as their advantage and disadvantage were summarized. RESULTS AND CONCLUSION:A total of 576 articles were included. Among them, articles published earlier, duplicated, and similarly were excluded, and 53 articles were finaly included. Various animal models of osteoporosis may only focus on the certain causes, certain stage, some of the main symptoms and some pathophysiological changes of disease. Accordingly, appropriate modeling methods and experimental animals should be selected based on research objective. Rat undergoing castration is the most commonly used model in the modeling of osteoporosis. Among drug methods for constructing osteoporosis model, glucocorticoids is the most commonly used one. Disuse method and nutritional method have limitations, and always combined with castration and drug methods. The effects of gene transfer, gene mutation and brain-derived model deserve
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Objective To explore the safety and short?term efficacy of irreversible electroporation (IRE)ablation which is a novel ablation technology in unresectable hepatic neoplasms. Methods Patients with pathologically diagnosed as liver cancer or liver metastases were prospectively enrolled. The patients were not suitable for surgery with PS score ≤ 2. Exclusion criteria included who was not tolerate general anesthesia, severe liver and kidney dysfunction, and with cardiac pacemaker. A total of 16 patients were included in this study. There was 12 males and 4 females, aged 40 to 86 years with mean age (60 ± 10)y. Ultrasound and CT guided percutaneous IRE ablation was performed. Perioperative hemodynamic changes were reviewed. Liver and kindey function before and 7 d after ablation was compare by t test. The adverse reactions within 30 d after ablation treatment were recorded. CT and MR scans within 1 month were performed and the 30 d curative effect was evaluated by the modified RECIST criteria. Results All patients received IRE treatment successfully, and some patients experienced adverse reactions within 30 days after ablation, including abdominal pain in 7 cases, peritoneal effusion in 5 cases, hydrothorax in 4 cases, fever in 3 cases, cough, nausea and vomiting in 2 cases, biliary tract infection and thrombocytopenia in 1 case. After symptomatic treatment, these symptoms were improved. Severe complications, such as massive haemorrhage and bile leakage didn't occur. At 30 days after ablation, the curative effects were evaluated. Complete response (CR) was achieved in 1 patient , partial response (PR) was achieved in 12 patients, stable disease (SD) was in 2 patients , and progressive disease(PD) was 1 patients . The tumor relief rate (complete response+partial response) was 81.3%. Conclusions IRE ablation in the treatment of unresectable hepatic malignant tumor could have many advantages, including high safety, mild adverse reactions, and short?term efficacy. However, its long?term effect still need further observation.
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BACKGROUND:Surgical treatment is commonly used for decompressing the spinal cord in multilevel cervical spondylotic myelopathy, but the optimum anterior cervical reconstructive method has not been determined. OBJECTIVE:To compare and analyze the biomechanical characteristics of four anterior cervical reconstructive techniques in the surgical management of multilevel cervical spondylotic myelopathy utilizing the three-dimensional finite element model. METHODS:A three-dimensional finite element model of an intact C2–7 segment was developed and validated based on healthy adult male CT images. Four additional models were developed from the fusion model, including anterior cervical discectomy and fusion, anterior cervical corpectomy and fusion, anterior cervical hybrid decompression and fusion and anterior cervical discectomy and fusion with Cage alone. Von Mises stresses on the plate and the disc of adjacent levels (C2/3, C6/7) were comparatively analyzed. RESULTS AND CONCLUSION:(1) The C2/3 disc stress in flexion, extension, lateral bending and rotation condition was significantly higher than the C6/7 disc in 4 anterior cervical reconstructive techniques, and the adjacent disc stress (C2/3, C6/7) was lowest in the anterior cervical discectomy and fusion with Cage alone, and highest in the anterior cervical corpectomy and fusion. (2) The stress at the plate-screw interface was highest in the anterior cervical corpectomy and fusion, and lowest in the anterior cervical discectomy and fusion. (3) In conclusion, among the four anterior cervical reconstructive techniques for multilevel cervical spondylotic myelopathy, the anterior cervical discectomy and fusion with Cage alone makes little effect on the adjacent disc stress, which might reduce the incidence of adjacent segment disease after fusion. However, the accompanying risk of the increased incidence of cage subsidence should never be neglected.
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BACKGROUND:Adjacent segment disease is a long-term complication of anterior cervical decompression and fusion, and has been paid great attention recently. Cause of disease contains increased range of motion in surgery adjacent segment, intervertebral height loss and high intradiscal pressure. In the clinic, compared with fusion surgery, whether cervical non-fusion can effectively decrease the incidence of adjacent segment disease remains poorly understood. OBJECTIVE:To systematicaly assess the effects of cervical fusion and cervical non-fusion on adjacent segment degeneration. METHODS:We retrieved the randomized controled trial concerning cervical fusionversuscervical non-fusion to repair cervical syndrome in the Medline, PubMed, Embase and Cochrane databases from January 2000 to December 2013. This study compared the effects of two repair methods on adjacent segment disease and evaluated methodological quality of the included studies. A meta-analysis was performed using RevMan 5.2 software. Outcome indicators: second surgery was undergone due to postoperative adjacent segment disease. RESULTS AND CONCLUSION: After screening, five randomized controled trials met the inclusion criteria. There were 1 602 patients. Al patients received surgery due to cervical syndrome. 785 cases underwent anterior decompression and fusion, and 817 cases underwent intervertebral disc replacement. 1 066 patients completed al folow-up, with a total folow-up rate of 66.54%. There were 494 patients treated with anterior decompression and fusion and 572 patients with intervertebral disc replacement. A total of 68 patients underwent second surgery due to adjacent segment disease. Total second surgery rate was 6.38% (68/1 066), including 28 cases after intervertebral disc replacement and 40 cases after anterior decompression and fusion. The grade of quality evaluation of the included studies was high, including five studies with grade A. Moreover, heterogeneity was smal (I2 = 14%). Meta-analysis results revealed that the second surgery rate of adjacent segment disease was lower after cervical non-fusion than after cervical fusion, which showed significant differences (odd ratio = 0.58, 95% confidence interval: 0.35, 0.96). These results suggested that the second surgery rate of adjacent segment disease was higher after cervical fusion than after cervical non-fusion. The alterations in cervical biomechanics caused by fusion greatly affected the occurrence of adjacent segment disease.
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Penicillium decumbens T. is an important filamentous fungus for the production of cellulases to effectively degrade lignocellulose for second generation biofuel production. In order to enhance the capability of Penicillium decumbens to produce cellulases, we constructed a creB (a deubiquitinating enzyme encoding gene) deletion cassette, and generated a creB knockout strain with homologous double crossover recombination. This mutation resulted in a detectable decrease of carbon catabolite repression (CCR) effect. The filter paper activity, endoglucanase activity, xylanase activity and exoglucanase activity of the deltacreB strain increased by 1.8, 1.71, 2.06 and 2.04 fold, respectively, when comparing with the parent strain Ku-39. A 2.68 fold increase of extracellular protein concentration was also observed. These results suggest that the deletion of creB results in CCR derepression. These data also suggest that CREB influences cellulase production of Penicillium decumbens. In generation, this study provides information that can be helpful for constructing cellulase hyper-producing strain.
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Celulase , Endopeptidases , Genética , Metabolismo , Técnicas de Inativação de Genes , Lignina , Metabolismo , Proteínas Mutantes , Metabolismo , Penicillium , Genética , Recombinação Genética , Proteínas Ubiquitinadas , Genética , UbiquitinaçãoRESUMO
Objective:To observe the up-regulation of nuclear Clusterin (nCLU)gene on the biological behaviors of human non-small cell lung cancer cell line A549 .Methods:Sense eukaryotic expression vector of nCLU was constructed by cloning the cDNA of nCLU into pIREShyg3 vector. A549 cells were transfected with pIRES-nCLU and pIREShyg3 vectors by lipofectAMINE~(TM) 2000 mediation, respectively. Stable transfected cells were selected by hygromycin B screening. CCK-8 assay was performed to evaluate the effect of nCLU over-expression on cell proliferation in vitro. The expression level of nGLU protein was examined by Western blotting. Cell cycle distribution was detected by FCM with PI staining. The alteration of migration and metastasis potential of A549 cells before and after nCLU gene transfection were assayed by cell chemotactic migration and invasion test. Results:The proliferation speed of the transfected A549 cell clones stably over-expressing nCLU was slowed down. FCM analyses revealed that the percentage of cells in G_0/G_1 phase dramatically increased from (33.54±2.10)% to (63.31±4.30)%. The cell chemotactic migration and invasion potentials were markedly reduced after nCLU gene transfection (P<0.05). Conclusion:Up-regulation of nCLU can greatly inhibited the proliferation and decreased the migration and invasion capabilities of A549 cells.
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Objective To investigate the dynamic expression and clinical significance of toll-like receptor 4(TLR4)/CD80 in patients before and early after renal transplantation. Methods Thirty-two patients who received renal transplantation were enrolled in his study. All of them were primary recipients, and the level of panal reactive antibody less than 1%. The expression of TLR4 and CD80 in CD14, positive monocyte of peripheral blood from atients on the 1st, 4th, 7th, 14th, 21st, 28th and 35 th days after transplantation were measured by three-color fluorescent taining flow cytometry. The pa-tients were divided into rejection roup(7 cases) and non-rejection group(25 cases) according to rejec-tion episode record within 2 weeks. Normal control group(10 cases) enrolled healthy adult volunteers. Diagnosis of acute rejection depended on clinical symptoms, lab test, color doppler onography and re-nal biopsy. Results Before ransplanstation, the ressions of TLR4 and CD80 were (8.03± 0.84)% and (0.85±0.31)% in rejection group, (6.14±0.85)% and (0.84±0.39)% in non-rejec-tion group, (6.37±0.56)% and (0.85±0.35)% in normal control group. The expression of TLR4 of rejection group was higher than those of non-rejection group and normal control group(P<0.01). The expressions of CD80 of 3 groups had no significant difference(P=0.995). After ransplanstation, the expressions of TLR4 and CD80 increased to (16.50±1.02)% and (7.82±1.66)% in rejection group, (11.60±0.98)% and (2.26±0.96)% in non-rejection group at the 4th day, and reached the peak levels at the 7th day: (36.40±4.86)% and (9.53±1.97)% in rejection group, (22.70± 3.45)% and (1.87±0.72)% in non-rejection group, then displayed downtrend, rejection group de-creased to (7.10±0.82)% and (0.87±0.57)% at the 35th day, non-rejection group decreased to (7.20±0.76)% and (0.81±0.37)% at the 21st day. Compared with non-rejection group, rejection group showed higher peak expression value of TLR4/CD80 (P<0.01) and longer lasting time. Con-clusions The high expression of TLR4 may increase the risk of acute rejection. The up-regulated TLR4/CD80 levels early after renal transplantation may ontribute to the happeness of acute rejection.
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The purpose is to obtain optimal cultural conditions for Ganoderma lucidum in the solid medium mainly containing the residual of Pteridium aquilinum,and to provide theoretical foundation for the further use of this material from Pteridium aquilinum.Response surface methodology was applied to optimize the main culture conditions including proportion of the residual compound in medium,water content in medium and culture temperature.The results showed that proportion of residual of Pteridium aquilinum in medium,water content in medium and cultural condition had noticeably significant effects on daily average growth rate of mycelium of Ganoderma lucidum(p