Inhibitory effect of 5-Fluorouracil on the proliferation of human osteosarcoma cells in vitro.

PURPOSE: To evaluate the inhibitory effect of 5-fluorouracil (5-FU) in combination with cisplatin on the proliferation of human osteosarcoma cells in vitro. METHODS: Three groups of human osteosarcoma U2OS cells were cultured in DMEM medium supplemented with the following drugs: 20 µg/mL 5-FU, 50 µg/mL 5-FU and 50 µg/mL 5-FU in combination with 0.5 mg/L cisplatin. After culture at 24, 48 and 72 h, the inhibition of proliferation rate of U2OS cells was calculated by CCK-8 cell kit. Cell invasiveness was assessed by Transwell assay. Flow cytometry was used for monitoring the cell apoptosis. RESULTS: 5-FU inhibited the growth of osteosarcoma cells, and the results of different concentrations of 5-FU were significantly different. The growth of U2OS cells decreased significantly within 24-72 h, and the concentration of 5-FU increased with time. The inhibition of the shift was more obvious, and the combined drug inhibition was significantly higher than the 20µg/ml 5-FU Group, 0.5mg/L Cisplatin Group and 50µg/ml 5-FU Group. After 72 h, the mean inhibitory rates of 20 µg/mL 5-FU, 50 µg/mL 5-FU, 50 µg/ml 5-FU in combination with 0.5 mg/L cisplatin, and 0.5 mg/L cisplatin were 12.54±1.26%, 22.17±0.59%, 32.54±1.25%, 20.84±0.83% respectively, and the difference was significant (p<0.05). Results of cell invasion assay showed that after culturing for 48 h, the mean number of cells penetrating the membrane was 22.84±5.27 in the culture group of 0.5 mg/L cisplatin, 30.57±5.68 in the culture group of 20 µg/mL 5-FU, 18.68±4.88 in the culture group of 50 µg/mL 5-FU, and 9.84±3.64 in the culture group of 50 µg/mL 5-FU in combination with 0.5 mg/L cisplatin, respectively, and 72.00±7.52 in the control group, showing statistical differences in each group (p<0.05). The apoptosis of the control group was significantly lower than that of the other groups (p<0.05). Apoptosis rate of the 20µg/mL 5-FU group was significantly lower than that of the 0.5mg/L cisplatin group, the 50µg/mL 5-FU group and the 50 µg/ml 5-FU group in combination with 0.5mg/L cisplatin (p<0.05). There was no difference in apoptosis between 0.5mg/L cisplatin and 50µg/mL 5-FU group (p>0.05). CONCLUSION: 5-FU in combination with cisplatin exerts an inhibitory effect on the proliferation and invasion of human osteosarcoma cells in vitro, and can promote cell apoptosis.

Prognostic value of the Epstein-Barr virus and tumor suppressor gene p53 gene in nasopharyngeal squamous cell carcinoma.

AIMS AND METHODS: Retrospectively, this paper compared the differences of the Epstein-Barr virus (EBV)-encoded small RNAs (EBERs), protein expression and gene mutations of tumor suppressor gene p53 (TP53) in keratinized nasopharyngeal squamous cell carcinoma (KNSCC) and nonKNSCC, and the relationships between pathological features and the prognosis of patients were analyzed. RESULTS: The positive rate of EBERs hybridization and TP53 expressions was 76.3% and 52.2%, respectively, while the mutation rate of TP53 gene was 39.6%. Logistic regression analysis showed direct relationships between the subtypes of nasopharyngeal squamous cell carcinoma (NPSCC) and EBERs-positive, or frequent consumption of pickled food. Overall survival rates of patients with positive TP53 expression, the TP53 gene mutations, vascular invasions, organ metastases, lymph node metastasis, and clinical recurrence were significantly lower than those of patients without those symptoms. The poorer prognosis was related to regularly drinking and the advanced age. According to the Cox regression analysis, we found that the main prognostic factors of NPSCC patients were the aging, recurrence, TP53 gene mutations, especially exon 7 or 8 mutations. CONCLUSIONS: We concluded that there were the correlations between NPSCC subtypes with EBV infection and frequent intaking of pickled food, while aging, clinical recurrence, and TP53 gene mutations were independent predictors for the poor prognosis of nasopharyngeal carcinoma.

Chondroid gastrointestinal stromal tumor in the stomach with early adenocarcinoma.

OBJECTIVE: To explore the pathologic features of gastric chondroid gastrointestinal stromal tumors. METHODS: The clinicopathologic data of one case of gastric chondroid gastrointestinal stromal tumor were collected and the features were analyzed by literature review. RESULTS: The male patient was 64 years old and had suffered from upper abdominal fullness discomfort without obvious cause for 5 years. Gastroscopic examination showed a rough area located in the lesser curvature of the gastric antrum, measuring 6 cm × 4 cm. CT scan showed the stomach wall was unevenly thick at the gastric antrum and stomach outlet. Multiple enlarged lymph nodes were seen nearby. The biopsy pathology showed adenocarcinoma of gastric antrum. The patient underwent laparoscopic gastrectomy and gastric chondroid gastrointestinal stromal tumor was found with adenocarcinoma of the stomach. Asp842Val mutation was found in the PDGFRα 18 exon. CONCLUSION: Gastric chondroid gastrointestinal stromal tumors are rare and low risk. Tumor cells express CD117 and Asp842Val mutation in the PDGFRα 18 exon revealed by genetic sequencing suggesting this kind of tumor might be resistant to imatinib.

Comparison of 2 femoral tunnel drilling techniques in anterior cruciate ligament reconstruction. A prospective randomized comparative study.

BACKGROUND: To evaluate the length and position of femoral tunnel,and exam whether knee stability and clinical functional outcomes are superior in AMP method. METHODS: From August 2014 to February 2015, we prospectively recruited 104 patients undergoing anterior cruciate ligament reconstruction. They were randomized to anteromedial portal or transtibial method. All patients underwent Lysholm score, International Knee Documentation Committee score,Tegner score at pre-operative and last follow-up point as subjective assessment of clinical function. The Lachman test, the Pivot-shift test and KT-1000 were performed at the last follow-up as a evaluation of knee joint stability. We measured the length of femoral tunnel intraoperatively and at 1 week post-operatively, CT-based three-dimensional reconstruction was used to assess femoral tunnel location. RESULTS: The average follow-up time of anteromedial portal group was 25.7 ± 6.8 months (range:12-36.5 months), and the average follow-up time of the transtibial group was 24.9 ± 6.0 months (range:12-37 months). There was no significant difference between the groups pre-operative Lysholm score, IKDC score and Tegner scores. Both groups showed significantly improvement in these clinical function scores at follow up for their ACL reconstruction. However, there was no significant difference in the function scores between the two groups at last follow up. However, the mean femoral tunnel length in the anteromedial portal group was significantly shorter than that in the transtibial group. And tunnel location was significantly lower and deeper with the anteromedial portal technique than with the transtibial technique. CONCLUSION: The use of anteromedial portal method resulted in a significantly lower and deeper placement of the femoral tunnel, and a shorter tunnel length compared to the transtibial method. However, there was no statistical difference in terms of clinical function and knee joint stability between the anteromedial portal method and the transtibial method. TRIAL REGISTRATION: Name of the registry: Chinese Clinical Trial Registry. The registration number: ChiCTR1800014874 . The date of registration: 12 February, 2018. The study is retrospectively registered.

Analyzing EGFR mutations and their association with clinicopathological characteristics and prognosis of patients with lung adenocarcinoma.

Epidermal growth factor receptor (EGFR) is an important gene in the development of lung adenocarcinoma. However, there is controversy regarding the association between EGFR mutations and survival time of patients with lung adenocarcinoma. In the present study, tissue specimens and clinical data were collected from 219 patients with lung adenocarcinoma who had not undergone prior radiotherapy or chemotherapy. EGFR mutations were detected using a fluorescence polymerase chain reaction method, and the association between EGFR mutations and clinicopathological characteristics was analyzed. Overall survival (OS) curves were constructed using the Kaplan-Meier method and the influence of clinicopathological characteristics on OS was analyzed using the Cox regression model. The EGFR mutation rate was 50.7%, and the most common mutations were the L858R substitution mutation in exon 21 (L858R; 54.9%) and the deletion mutation in exon 19 (19-Del; 36%). The presence of EGFR mutations varied significantly with sex, smoking history, T stage, vascular invasion and adenocarcinoma subtypes (P<0.05). The survival time was significantly longer for female, young (<60 years-old), non-smokers or patients exhibiting EGFR mutations (G719X, 19-Del, L858R and L861Q). The survival time was also significantly longer for patients with a 19-Del mutation, early stage tumors, tyrosine kinase inhibitors targeted therapy-treated patients, for those not exhibiting nerve or vascular invasion, and for those without disease recurrence (P<0.05). Multivariate analysis revealed that tumor pathological Tumor-Node-Metastasis (pTNM) stage, nerve invasion, vascular invasion, EGFR mutation and the 19-Del mutation were independent predictors (P<0.05). Therefore, tumor pTNM stage, nerve invasion, vascular invasion and EGFR mutation status, particularly that of 19-Del, were independent prognostic factors for patients with lung adenocarcinoma.

A study on role of triiodothyronine (T3) hormone on the improvement of articular cartilage surface architecture.

The present study was aimed to investigate the effect of triiodothyronine (T3) on the improvement of articular cartilage surface architecture at in vitro level. The T3 hormone was applied to neo-tissues in the range of 50, 100, 150 and 200ng/ml for 5 weeks. At the end of the treatment, biochemical and histological evaluation was carried out in the neo-tissues. T3 hormone application significantly increased the collagen production in neo-cartilage tissues. The properties of tensile and compressive were significantly increased compared to the controls. However, T3 hormone application also induced hypertrophy. At the higher dose concentration of T3 hormone application, tensile and compressive properties were tremendously increased 4.3 and 4.6 fold respectively. Taking all these data together, it suggested that the T3 hormone application could be a potential agent to increase the functional properties such tensile and compressive in neo-tissues.

miR-22 promotes apoptosis of osteosarcoma cells via inducing cell cycle arrest.

To study the effects of miR-22 on the proliferation and the apoptosis of osteosarcoma MG-63 cell line and to explore the potential molecular mechanism that miR-22 regulates this biological process. Quantitive real-time polymerase chain reaction (RT-qPCR) was performed to explore the miRNA level of miR-22. The MG-63 cell line was infected with miR-22 mimics for establishment of miR-22 overexpression. Non-infected cells were in blank group and cells infected with empty vector were served as negative control (NC group). MTT assay was conducted to measure cell viability. The cell cycle and apoptosis were explored using flow cytometry and the apoptosis-related markers were detected by western blotting. RT-qPCR results revealed that the miR-22 miRNA level in the MG-63 cells was significantly lower than that in osteoblasts (P<0.05). MTT assay showed that the MG-63 cells infected with miR-22 mimics exhibited markedly decreased proliferation ability compared with blank and empty vector (NC) groups. Next, we found that overexpression of miR-22 remarkably increased the apoptosis of the MG-63 cells, evidenced from the flow cytometry results and elevated Bax and reduced Bcl-2. Furthermore, results revealed that percentage of the cells at G0/G1 phase in miR-22 mimic group (66.75±3.67%) was significantly higher than blank (52.9±2.58%) and NC (50.5±2.45%) groups. miR-22 attenuated the proliferation and induced the apoptosis of the MG-63 cells via promoting G0/G1 cell cycle arrest. Thus, miR-22 may have the potential to be a novel therapeutic in treatment of osteosarcoma.

Hypocalcaemia After Total Knee Arthroplasty and its Clinical Significance.

BACKGROUND: Transient hypocalcaemia is a frequent complication after total knee arthroplasty (TKA). In this study, we investigated the factors associated with the development of hypocalcaemia after TKA in order to explore its clinical significance and treatment. PATIENTS AND METHODS: A retrospective analysis of the change of serum calcium levels for 40 patients after TKA was performed. We investigated the patients prospectively for age, gender, and amount of bleeding at operation. At 24 hours following the operation, serum calcium of the patients was evaluated and a t-test was performed to analyze the categorical variables. Pearson's correlation analysis was used to determine the risk of hypocalcaemia in univariate analysis. RESULTS: After TKA, the serum calcium level was significantly lower than that before the operation (p<0.01); the incidence of postoperative hypocalcaemia was 77.5%, the decline was positively correlated with intraoperative blood loss (Pearson's r=0.405, p=0.01). CONCLUSION: Hypocalcaemia occurs frequently after TKA, however, clinical symptoms associated with hypocalcaemia are rare. The calcium ion is an important electrolyte, neurotransmitter and blood coagulation factor. It is suggested that we should routinely monitor calcium ion levels during the perioperative period and deal with hypocalcaemia in a timely fashion.

Shikonin protects chondrocytes from interleukin-1beta-induced apoptosis by regulating PI3K/Akt signaling pathway.

Chondrocyte apoptosis is mostly responsible for the development and progression of osteoarthritis. IL-1ß is generally served as an agent that induces chondrocyte apoptosis. Shikonin exerts its anti-inflammatory effect on cartilage protection in vivo. We aimed to explore the protective effect of shikonin on interleukin-1beta (IL-1ß)-induced chondrocyte apoptosis and the potential molecular mechanisms. Chondrocytes were isolated from the joints of newborn Sprague-Dawley rats. The MTT assay and LDH cell death assay were used to determine the cell viability and chondrocyte apoptosis was detected by Annexin-V/PI staining and nucleosomal degradation. The contents of phosphorylated-PI3K (p-PI3k), phosphorylated-Akt (p-Akt), Bcl-2, Bax, and cytochrome c were detected by Western blotting. A quantitative colorimetric assay was used to detect the caspase-3 activity. Our results showed that pretreatment with shikonin (4 µM) inhibited cytotoxicity and apoptosis induced by IL-1ß (10 ng/ml) in chondrocytes. Shikonin pretreatment also decreased the activity of IL-1ß that decreased Bcl-2 expression and levels of p-PI3K and p-Akt, and increased Bax expression, cytochrome c release, and caspase-3 activation. It also reversed the activity of IL-1ß that promoted the synthesis of matrix metalloproteinase-13 and inhibited the expression of tissue inhibitor of metalloproteinase-1 expression, with the net effect of suppressing extracellular matrix degradation. These data suggested that shikonin may protect chondrocytes from apoptosis induced by IL-1ß through the PI3K/Akt signaling pathway, by deactivating caspase-3.

[A study of three different procedures for avulsion fracture of tibial intercondylar eminence].

OBJECTIVE: To observe the clinical outcomes and differences among three surgical procedures for avulsion fracture of tibial intercondylar eminence. METHODS: From October 1995 to October 2005, 3 different procedures had been performed on 49 patients, which included open reduction and internal fixation (Group A, n = 17), arthroscopic reduction and internal fixation( Group B, n = 19) and limited open reduction and internal fixation assisted with arthroscopy (Group C, n = 13). All patients were followed up for 1 to 10 years (average 4. 6 years). RESULTS: For the 3 groups, normal extension function were 35.5%, 16.0% and 38.0%, mild abnormal 35.5%, 11.0% and 23.0%, moderate abnormal 29.0%, 47.0% and 31.0%; and severe abnormal were 0,26.0% and 8.0%. Normal flexion were 82.0%, 78.0% and 84.0%, mild abnormal 12.0%, 11.0% and 8.0%, moderate abnormal 6.0%, 11.0% and 8.0%. The positive rate of Lachman's or anterior drawer test were 35.0%, 45.0% and 38.0%; McIntoshi test were 11.0%, 16.0% and 13.0% respectively for 3 groups. Lysholm' scale were average 98.6, 97.3 and 98.2; Tegner' scale were 6.6, 6.4 and 6.7. KT-2000 showed that anterior translation of tibial were 6.9, 7.1 and 6.6 mm; side to side difference were 11.4, 1.7 and 1.5 mm, except that statistically significant differences were found in extension function between group A and group B (P = 0.02). There were no any statistically significant differences in other aspects. CONCLUSION: Limited open and properly over reduction and three dimensional as well as strong internal fixation assisted with arthroscopy should been attempted for the treatment of avulsion fracture of tibial intercondylar eminence.