Follow-up study on autogenous osteochondral transplantation for cartilage defect of knee joint / 中国骨伤

OBJECTIVE@#To evaluate clinical effect of autologous osteochondral transplantation in treating localized knee cartilage defects.@*METHODS@#Fifteen patients with knee cartilage defects were treated by autologous osteochondral transplantation from January 2007 to January 2008, including 8 females and 7 males, aged from 23 to 45 years old. Preoperative and postoperative KSS score at 10 years were compared.@*RESULTS@#All patients were followed up for 10.0 to 10.7 years, with an average of(10.2±0.3) years. Clinical score of KSS was improved from 38.86±4.09 to 85.07±2.19 at 10 years after operation(0.05). All patients had no other complications.@*CONCLUSIONS@#Through long-term follow-up of patients with cartilage defect in knee treated by autologous bone cartilage transplantation showed that this method could effectively improve function of knee joint and alleviate pain. So it is an effective method for repair of osteochondral defect.

Role of 8-allyl Garcinol in the Chemoprevention of Oral Squamous Cell Carcinoma / 中国医学科学院学报

Objective To evaluate the chemopreventive effects of 8-allyl garcinol on oral squamous cell carcinoma(OSCC).Methods OSCC cell line CAL27 were cultured and treated with different concentrations of garcinol or 8-allyl garcinol. Their effects on the biological behaviors of OSCC cell line CAL27 were measured by MTT assay,clony formation assay,scratch migration assay,and flow cytometry with Annexin V-FITC/PI staining assay. We established DMBA-induced hamster cheek pouch models of dysplasia. While the negative control group was not treated,the positive group was treated with 0.5% DMBA solution tropically to the left cheek pouch three times per week for three consecutive weeks. The other four groups received 0.5 mmol/L or 1.0 mmol/L garcinol or 8-allyl garcinol respectively three times within the following two weeks after DMBA treatment. Hamsters were sacrificed at the fifth week to obtain tissue samples of the left cheek pouch. The samples were examined by histopathology and BrdU immunohistochemisty.Results MTT assay showed that both garcinol and 8-allyl garcinol inhibited the proliferation of CAL27 cells in a concentration-and time-dependent manner. The half maximal inhibitory concentration(IC)of 8-allyl garcinol[(13.13±2.55)μmol/L] was significantly lower than garcinol[(32.20±3.24)μmol/L;t=8.008,P=0.001]. Comparing the two grougs of medicine in the same concentration,the inhibiting proliferation effects 8-allyl garcinol had significantly stronger effect in inhibiting proliferation than garcinol when the same dose was applied,and the difference was largest at the concentrations of 10(24 h:t=8.012,P=0.001;48 h:t=5.939,P=0.001;72 h:t=12.551,P=0.001)and 20 μmol/L(24 h:t=8.887,P=0.001;48 h:t=9.324,P=0.002;72 h:t=5.361,P=0.002). The clone formation assay showed the clone formation rates after the treatment with 20 μmol/L garcinol and 20 μmol/L 8-allyl garcinol were(44.1±0.4)% and(23.6±0.6)%,respectively,which were significantly lower than those after treatment with 10 μmol/L garcinol[(55.6±2.8)%;t=6.894,P=0.019] and 10 μmol/L 8-allyl garcinol[(31.0±0.6)%;t=15.556,P=0.001]. The inhibiting effects of 8-allyl garcinol at the concentrations of 10 μmol/L(t=14.682,P=0.003)and 20 μmol/L(t=51.514,P=0.001)were significantly stronger than garcinol.Scratch test showed the relative cell migration rates after treatment with 10 and 20 μmol/L garcinol for 12 hours were(16.00±4.55)%(t=3.139,P=0.026)and(3.00±3.16)%(t=6.608,P=0.001),respectively,which were lower than negative control [(30.33±7.64)%]. The relative cell migration rates after treatment with 10 and 20 μmol/L 8-allyl garcinol for 12 hours were(16.25±3.86)%(t=3.245,P=0.023)and(6.00±2.65)%(t=5.214,P=0.006),respectively,which were also lower than negative control[(30.33±7.64)%]. In addition,the relative cell migration rates after treatment with 10 and 20 μmol/L garcinol for 24 hours were(23.75±4.57)%(t=4.718,P=0.005)and(5.75±1.50)%(t=10.432,P=0.001),respectively,which were lower than negative control[(45.33±7.64)%]. The relative cell migration rates after treatment with 10 and 20 μmol/L 8-allyl garcinol for 24 hours were(23.50±2.38)%(t=5.529,P=0.003)and(11.67±2.31)%(t=7.308,P=0.002),respectively,which were also lower than negative control[(45.33±7.64)%]. Furthermore,the relative cell migration rate after treatment with 20 μmol/L garcinol for 24 hours was significantly lower than after treatment with 8-allyl garcinol(t=4.151,P=0.009). The apoptosis experiments showed that the early apoptosis rate of CAL27 cells was(5.00±0.10)% after treatment with 10 μmol/L garcinol,which was significantly higher than negative control[(1.57±0.21)%;F=70.950,P=0.001]. The early and late apoptosis rates of CAL27 cells were(5.90±0.78)%(t=39.384,P=0.001)and(9.73±1.67)%(t=10.101,P=0.001),respectively,after treatment with 20 μmol/L garcinol,which were also significantly higher than negative control. The early apoptosis rate of CAL27 cells was(4.63±1.16)% after treatment with 8-allyl garcinol,which was significantly higher than negative control(t=4.511,P=0.041). The effects of 8-allyl garcinol in promoting cell apoptosis were weaker than garcinol(10 μmol/L:t=5.982,P=0.004;20 μmol/L:t=8.578,P=0.001). The histopathological test also showed that the hyperplastic areas of oral mucosal epithelium in hamsters after treatment with 0.5 mmol/L garcinol(t=2.546,P=0.031),0.5 mmol/L 8-allyl garcinol(t=3.485,P=0.008),1.0 mmol/L garcinol(t=4.556,P=0.001),and 1.0 mmol/L 8-allyl garcinol(t=5.393,P=0.001)were significantly smaller than positive control. The dysplasia areas of oral mucosal epithelium in hamsters after treatment with 0.5 mmol/L 8-allyl garcinol(t=2.130,P=0.046),1.0 mmol/L garcinol(t=3.434,P=0.010),and 1.0 mmol/L 8-allyl garcinol(t=4.518,P=0.004)were also smaller than positive control;1.0 mmol/L garcinol group(t=2.793,P=0.023)and 1.0 mmol/L 8-allyl garcinol group(t=4.997,P=0.001)were smaller than 0.5 mmol/L garcinol treatment group. Immunohistochemical staining of BrdU showed that the BrdU-labeled indicators were significantly lower in negative control group(t=7.563,P=0.001),0.5 mmol/L garcinol(t=2.862,P=0.029),0.5 mmol/L 8-allyl garcinol(t=4.693,P=0.002),1.0 mmol/L garcinol(t=5.071,P=0.002),and 1.0 mmol/L 8-allyl garcinol(t=5.133,P=0.001)when compared with the positive control. The BrdU-labeled indicators in 0.5 mmol/L 8-allyl garcinol(t=3.724,P=0.007),1.0 mmol/L garcinol(t=7.000,P=0.001),and 1.0 mmol/L 8-allyl garcinol(t=4.413,P=0.003)were also significantly lower than in 0.5 mmol/L garcinol group.Conclusions 8-allyl garcinol could inhibit the proliferation and migration of OSCC cell line CAL27 and promotes apoptosis. It also has prominent inhibitory effects on DMBA-induced hamster cheek pouch dysplasia. However,the specific effects are slightly different from garcinol.

SIRT6 regulating IGF2 though recruiting mRNA binding protein IMP1 in 293T cells / 基础医学与临床

Objective To investigate the post-transcriptional regulation mechanism of insulin-like growth factor 2 (IGF2),which is an important tumor regulator,by SIRT6. Methods SIRT6 was overexpressed in 293T cells and IP was used to enrich SIRT6 and mass spectrometry(MS)was used to detect the protein that interacted with SIRT6. Western blot was used to validate the interacted protein and its acetylation/phosphorylation modification status. Results SIRT6 did not change the acetylation modification status of IMP1,but the phosphorylation status of IMP1 was elevated in the presence of SIRT6. Conclusions SIRT6 may regulate IGF2 though promoting the phosphoryla-tion of IMP1 in 293T cells.

Reconstruction of complex proximal tibial defects using the long-stem tibial component combined with metallic wedge / 中国骨伤

<p><b>OBJECTIVE</b>To investigate results of total knee arthroplasty using the long-stem tibial component combined with metallic wedge of knee prosthesis for the treatment of proximal defects.</p><p><b>METHODS</b>From January 2011 to May 2013, 10 patients (11 knees) were treated with total knee arthroplasties using the long-stem tibial component with metallic tibial wedge of knee prosthesis. All the patients were female and the average age was 67 years old (ranged, 60 to 77 years old). All the patients were osteoarthritis. All the patients were classified as T2A style. The patients were evaluated according to knee score system (KSS).</p><p><b>RESULTS</b>All the patients were followed up for 12 months on average (ranged 3 to 29 months). The clinical outcome was assessed using KSS score, including knee pain score, knee stability score, knee range of motion score and knee walking score, knee stairs score. There were significantly differences at 6 weeks, 3 months, 6 months and 12 months between pre-and postoperative KSS score.</p><p><b>CONCLUSION</b>The mechanical stability of tibial fixation in primary TKA is significantly increased by using the long-stem tibial component with metallic wedge of knee prosthesis, even in the presence of poor proximal bone.</p>

Efficacy of Topical Tacrolimus for Erosive Oral Lichen Planus: A Meta-analysis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To assess the efficacy and safety of topical tacrolimus for erosive oral lichen planus (EOLP).</p><p><b>METHODS</b>Literatures published up to December 2013 were searched from PubMed, Embase, CENTRAL, Chinese BioMedical Literature Database (CBM), and System for Information on Grey Literature in Europe (SIGLE). All randomized controlled trials (RCTs) of topical tacrolimus for EOLP which compared with other interventions or a placebo were considered in this Meta-analysis. Two researchers collected data independently. The assessment of methodological quality was based on Cochrane Handbook and the materials were analyzed with the software Revman 5.2.5. The primary outcome measures were the symptoms (e.g. pain, discomfort) complained by patients. The secondary outcome measures included the improvement rate of clinical signs assessed by the investigators and the incidence of adverse effects (e.g. clinical candidiasis).</p><p><b>RESULTS</b>A total of 9 RCTs involving 476 patients were finally included. The pooled odds ratio (OR) of clinical improvement for topical tacrolimus vs. topical corticosteroids was 1.19 [95% confidence interval (CI): 0.64-2.22, I2: 44%]. Regarding to 0.1% tacrolimus and 0.03% tacrolimus, the pooled OR were 1.87 (95% CI: 0.60-5.82) and 1.47 (95% CI: 0.14-16.04) respectively in subgroup analysis. No serious adverse events were reported in topical tacrolimus group.</p><p><b>CONCLUSIONS</b>There was no evidence to support that topical tacrolimus for EOLP was more effective and safer than topical corticosteroids in this Meta-analysis. Clinical assessment criteria should be established and accepted by clinicians and researchers before further RCTs are undertaken.</p>

Application of cDNA array for studying the gene expression profile of mature appressoria of Magnaporthe grisea / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Appressorium is an infection structure of the phytopathogenic fungus Magnaporthe grisea. Analysis of gene expression profiles of appressorium development provides insight into the molecular basis of pathogenicity and control of this fungal plant disease. A cDNA array representing 2927 unique genes based on a large EST (expressed sequence tag) database of M. grisea strain Y34 was constructed and used to profile the gene expression patterns at mycelium and appressorium maturation stages. Compared with mycelia, 55 up-regulated and 22 down-regulated genes were identified in mature appressoria. Among 77 genes, 16 genes showed no similarity to the genome sequences of M. grisea. A novel homologue of peptidyl-prolyl cis-trans isomerase was found to be expressed at low-level in mature appressoria of M. grisea. The results indicated that the genes such as pyruvate carboxylase, phospholipid metabolism-related protein and glyceraldehyde 3-phosphate dehydrogenase involved in gluconeogenesis, lipid metabolism and glycolysis, showed differential expression in mature appressoria. Furthermore, genes such as PTH11, beta subunit of G protein and SGT1 involved in cell signalling, were expressed differentially in mature appressoria. Northern blot analysis was used to confirm the cDNA array results.

Expression of NOS III mRNA in different tissues of spontaneously hypertensive rats using RNA array / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the expression of nitric oxide synthase III (NOS III) mRNA in the heart, aorta, kidney and liver of spontaneously hypertensive rats (SHR).</p><p><b>METHODS</b>Two hundred and ninety-four total RNA samples were obtained from the tissues of ventricle, aortic smooth muscle, kidney and liver of SHR and normotensive rats (Wistar-Kyoto rats, WKY). RNA array was used to determine the mRNA levels of NOS III of the two groups.</p><p><b>RESULTS</b>Compared with WKY, the systolic blood pressure increased significantly in SHR at 6-week-old, 8-week-old, 10-week-old and 12-week-old [(158.50 +/-7.69 vs 108.67 +/-5.89) mmHg, (174.33 +/-4.46 vs 128.50 +/-4.97) mmHg, (198.00 +/-13.45 vs 142.00 +/-3.58) mmHg, (216.67 +/-8.91 vs 141.17 +/-4.92) mmHg, P<0.01], and the ventricle/body weight ratio was significant higher at 10-week-old and 12-week-old [(4.08 +/-0.17 vs 3.59 +/-0.11, 4.05 +/-0.18 vs 3.40 +/-0.19)mg/g, P<0.01]. In the heart tissue and the kidney, the mRNA levels of NOS III were significantly increased at 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.12 +/-0.18 vs 0.90 +/- 0.15, 1.46 +/- 0.34 vs 1.06 +/-0.18, 1.66 +/- 0.31 vs 1.21 +/- 0.30, 1.98 +/- 0.40 vs 1.31 +/-0.38, P <0.05) and at 4-week-old, 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.10 +/- 0.21 vs 0.81 +/-0.11, 1.28 +/-0.18 vs 0.95 +/-0.13,1.31 +/-0.23 vs 0.99 +/-0.23, 1.70 +/-0.30 vs 1.08 +/-0.25, 1.83 +/-0.33 vs 1.15 +/-0.20, P<0.05 or P<0.01), respectively. There was no significant difference of the NOS III expression in the liver and no significant signals were detected in the aortic smooth muscle.</p><p><b>CONCLUSION</b>The results provide the evidence of the increased expression of NOS III in different tissues in SHR and suggests the progressive nature of essential hypertension.</p>