To investigate the role of Gli1 in DDP-resistance in lung cancer.Methods:A DDP-resistant human lung adenocarcinoma cell line A 549/DDP was cultured and transfected with Gli 1 siRNA.The mRNA and protein expression levels of Gli 1 were evaluated by qPT-PCR,Western blot analysis and immunofluorescence microscopy.The expression of Bcl-2、caspases-3、cyclinD1 was evaluated by Western blot analysis.Hoechst 33258 staining and flow cytometry were used to detect spontaneous cell apoptosis and cell cycle.The cell inhibition rate and DDP-induced cell death were examined by MTT and Annexin V-FITC/propidium iodide staining.Results:Gli1-knockdown by using Gli 1-specific siRNA led to a markedly decrease in Gli 1 mRNA and protein expression levels,when compared to negative siRNA transfected cells and untreated control cells (P=0.001).The downstream effectors of Gli1, Bcl-2 and cyclinD1 proteins were also inhibited (P=0.003),the expression of caspase-3 was increased (P=0.001).Hoechst 33258 staining showed that Gli1 depletion by Gli1 siRNA in A549/DDP cells could induce spontaneous apoptosis.The result of cell cycle showed Gli1 siRNA could lead more cells arrest in G1 phase.The IC50 of A549/DDP cells on DDP was (12.63 ±1.11 )μg/ml /(13.81±1.14)μg/ml,which decreased to (2.65±0.85)μg/ml after being transfected with Gli1 siRNA (P=0.000).Annexin V-FITC/propidium iodide staining showed that DDP-induced apoptosis rate in Gli 1-silencing cells was higher than that in negative siRNA or untreated control cells ( 35.19%±3.92% vs 6.43%±0.11%/5.01%±0.77%, P=0.000 ).Conclusion: Application of RNA interference can restrain the expression of Gli 1 mRNA and protein observably in A549/DDP cells,and increase the sensitivity of A549/DDP cells to cisplatin.Maybe Gli1 will become a new target to reverse the cisplatin resistance for lung cancer.

[Objective] To study the effect of arthroscopic reconstruction of anterior cruciate ligament using RIGIDfix and Intraflx system with quadruple-strand hamstring tendons.[Method]Thirty-six cases of anterior cruciate ligament ruptures were reconstructed arthroscopically with four strands of hamstring tendons,and the grafts were fixed with RIGIDfix and Intrafix system.Ptients were treated with systematic rehabilitation therapy and relevant complications were managed post-operatively.The patients were followed up for 6~12 months,ask the patients' subjective symptom,examine the knee range of motion,anterior drawer test,Lachman test and Pivot shift test,were managed to evaluate clinical result according to Lysholm rating.[Result]All patients had no serious complication,and were followed up for 6~12 months post-operatively.All patients had returned his post,knee range of flexion achieved 130?,extention achieved 0?.Three cases sometimes felt his knee soft and instabilization during exercise violently,the other patients had no sensation of soft and instabilization.When patients were followed up,anterior drawer test:negative 35 cases,positive 1 case;Lachman test:negative 33 cases,positive 3 cases;Pivot shift test of all patients were negative.The Lysholm score increased from 51.3?4.2 to 85.6?4.6,the difference was significant(P

Objective To explore the diagnostic value of quantitative contrast‐enhanced ultrasonography(CEUS) for crush injury in the hind limb muscle of rabbits .Methods A totle of 120 New Zealand white rabbits were randomized to receive compression on the left hind limb for either 2 h( n =56) or 4 h( n = 56) to induce muscle crush injury ,another 8 animals were not injured and served as normal controls .CEUS parameters such as peak intensity(PI) ,area under curve(AUC) ,ascending slop(AS) and descending slop(DS) were measured at 0 5. h ,2 h ,6 h ,24 h ,3 d ,7 d ,14 d after decompression .Results Compared with the uninjured muscle ,reperfusion of the injured muscles showed early and high enhancement in CEUS .The time‐intensity curve showed a trend of rapid lift and gradual drop .The PI and AUC values differed significantly among the three groups and were positively correlated with serum and tissue biomarkers .Rabbits of the 4 h compression group showed significantly higher PI and AUC values compared with the 2 h compression group at each time points .Conclusions CEUS can effectively detect muscle crush injury and monitor dynamic changes of the injured muscle in rabbits .PI and AUC are promising diagnostic parameters for this disease .CEUS might play an important role in the pre‐hospital and bedside settings for the diagnosis of muscle crush injury .

Immune checkpoint inhibitors significantly improves the prognosis of patients with advanced malignancy, but it is also associated with off-target toxicity caused by activation of the immune system, known as immune-related adverse events (irAEs). Severe irAEs will lead to temporary or permanent termination of immunotherapy, which greatly affects its clinical application. At present, glucocorticoids are mainly used to treat irAEs clinically. On one hand, severe adverse reactions will cause serious damage to patients' health; on the other hand, the extensive application of glucocorticoids will affect the efficacy of immune checkpoint inhibitors. In recent years, TNF-α inhibitors have shown significant effect in reducing toxic and side effects. This paper reviews the progress of TNF-α in preventing and treating irAEs.

BACKGROUND: A large amount of cell dies of apoptosis in secondary injury stage after spinal cord injury.OBJECTIVE: To probe into the influence of local intraspinal infusion with danshen injection on necrosis and apoptosis of spinal cell after acute spinal cord injury.DESIGN: Randomized controlled experiment was designed.SETTING: Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College.MATERIALS: Forty-four Chinese white rabbits of first grade were employed, aged varied from 4 to 5 months, of either sex, mass weighted varied from 2.0 to 2.5 kg.INTERVENTIONS: The experiment was performed in Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College from June 2002 to July 2003. Two groups were randomized, named danshen group and the control group, 22 rabbits for each. In both groups, modified Allen method was used to prepare the model of incomplete spinal cord injury. In danshen group, danshen injection was infused from subdural catheter for 4times at 0. 3 mL/kg per day of the total amount (once every 6 hours). In the control group, physiological saline of same dose was injected. The animals were sacrificed at the 8th, 24th and 72nd hours successively after injection for pathological and histomorphologic observation, peroxide dismutase and malondialdehyde determination and determination of positive cell count of B-cell lymphoma-2 (Bcl-2), the apoptosis-inhibition gene.MAIN OUTCOME MEASURES: Cell apoptotic index and cell apoptotic rate in the region of spinal cord injury.RESULTS: Forty-four rabbits entered result analysis for all. ① Results of cell apoptosis: Apoptotic index in danshen group was less remarkably than that in the control group(13.10 ± 1. 38, 20.39±2.96, 4.101, P ＜0.01); cell apoptotic rate was lower remarkably than the control group[ (9.67 ± 1.09)%,(14.68±2.81)%, t=4.072, P ＜0.01] and Bcl-2 expression was more than that in the control group[ (19. 12 ± 4.74) /mm2, ( 13.37 ± 3.68) /mm2,t = 2. 347, P ＜ 0.01 ]. ② POD content: The result in danshen group was higher thanthe control group[ (136.20 ± 13.64) NU/mL, (101.70 ± 15.24) NU/mL,t = 4. 132, P ＜ 0.01 ]. ③ Malondialdehyde content: The result in danshen group was lower than the control group[ (1.27 ± 0. 22) nmol/mL,(2.54±0.69) nmol/mL, t=4.309, P ＜0.01] . ④Degeneration and necrosis of neuron and neural fiber: The result in danshen group was milder than the control group.CONCLUSION: After local infusion with danshen(Radix Salviae Miltiorrhizae), cell apoptosis was decreased in local spinal cord injury and cell necrosis was inhibited and alleviated after acute spinal cord injury.

Objective To establish a mini pig model suitable for interventional studies in vivo. Methods The endothelia of unilateral renal arteries in 8 purebred Chinese experimental mini pigs(CEMP)was denuded by inflated balloons after the animals were fed with high cholesterol diet for 13 weeks.The CEMP were fed with h high cholesterol diet continuously till the 40th week.The levels of blood lipid panel and creatinine were tested at week 1,14 and 40.Bilateral renal arteries were examined with intravascular uhrasonography at week 14 and 40.The vessel samples were collected at week 40 and stained with haematoxylin-eosin,Masson trichrome technique, oil O and anti-macrophage immunohistological technique. Results Significant differences of blood lipid panel and creatinine were found between week 1 and week 40.Focal ischemic renal injury could be observed pathologically.Renal arteries of CEMP were suitable for interventional procedure such as angiography and intravascular ultrasonography.Cross-sectional information of vessels could be provided clearly by intravascular ultrasonography and the intimamedian thickness of injured renal arteries was much thicker than that of non-injured ones[(0.89±0.03)mm vs (0.30±0.02)mm,P＜0.05]as evidenced by this diagnostic technique.Pathological findings demonstrated the atheroselerotic profiles of the injured renal arteries.Fibrous and fibro-fatty plaques were the main pathologic types in this CEMP model. Conclusions An animal model with renal arterial atherosclerosis mimicking the progression of atheroselerotic renovaseular disease,which is suitable for interventional procedure is established successfully.Intravascular ultasonography may have potential clinical prospect on the evaluation of atherosclerotic renovaseular disease.

Objective To investigate the expression of glucose transporters 1 (GLUT-1) in gastric cancer and its relation with clinicopathological characteristics and prognosis. Methods PubMed, Web of Science,EMbase,Cochrane Library,WanFang Databases and China National Knowledge Internet(CNKI)were used to search literatures about GLUT-1 and gastric cancer. From the day of establishment to May 15, 2017, according to the inclusion and exclusion criteria, 2 researchers independently screened studies, extracted data and assessed quality of the included studies. Effect value and 95 % CI was calculated respectively. Then Meta-analysis was conducted by using RevMan5.3 software. Results A total of 11 articles were enrolled, including 1 714 cases in the gastric cancer group and 431 cases in the normal gastric mucosa group. The results of Meta-analysis showed that GLUT-1 expression was higher in the gastric cancer group than that in the normal group, and there was a significant difference (OR= 24.23, 95 % CI 11.86-49.51, P< 0.000 01). The expression of GLUT-1 was not related with age, gender, tumor size and invasion depth (all P> 0.05), but related with differentiated degree (OR= 0.41, 95 % CI 0.25-0.67, P= 0.000 4), lymphatic metastasis (OR=5.11, 95 % CI 2.73-9.56, P<0.000 1), and TNM staging (OR= 0.32, 95 % CI 0.20-0.51, P< 0.000 01). Moreover, GLUT-1 had a correlation with the overall survival rate of gastric cancer (HR= 1.61, 95 % CI 1.30-1.99, P < 0.000 1). Conclusions GLUT-1 protein expression is higher in gastric cancer tissues than that in normal gastric mucosa, and it is related to tumor differentiation degree, lymph node metastasis, TNM staging.Besides,GLUT-1 may be correlated with poor prognosis of patients with gastric cancer.

Objective To investigate the correlation of BRAF mutation frequency with cervical lymph node metastasis ,and to compare the ultrasonic characteristics in patients with BRAF mutation in papillary thyroid carcinoma ( PTC) different subtypes . Methods The tumor samples were collected from 139 PTC patients who underwent thyroidectomy . And they were classified by histological subtype into 3 groups:classic variant of papillary thyroid carcinoma (CVPTC) group( 34 cases) ,follicular variant of papillary thyroidcarcinoma (FVPTC) group(36 cases) ,tall cell variant (TCV) group(69 cases) . The BRAF mutation frequency and the correlation with cervical lymph node metastasis among 3 groups were analyzed , then the ultrasonic characteristics with BRAF mutation in PTC different subtypes were compared . Results①The frequency of BRAF mutation was statistically significant different in different subtypes( χ2 =6 .390 , P =0 .041) ,and the frequency in TCV was 86 .9％ . There was also a statistical difference between BRAF mutation frequency and cervical lymph node metastasis among three subtypes ( χ2 = 13 .106 , P =0 .041) .②There was no statistically significant difference among the three groups in nodule number ,echo level , internal structure ,boundary ,crossbar ,morphology and acoustic halo of patients with BRAF mutation ( P >0 .05) . ③ A single factor analysis was performed for the ultrasonographic characteristics of patients with BRAF mutation ,and there were significant statistical differences among the 3 groups in calcification type (χ2 = 21 .7 , P = 0 .001 ) and close to the envelope (χ2 = 7 .726 , P = 0 .021 ) . ④ Multivariate logistic regression showed that BRAF mutation was an independent influence factor affecting the calcification type of different histological subtypes in PTC patients.Conclusions ①BRAF mutation is correlated with cervical lymph node metastasis in different PTC subtypes . ② BRAF mutation is an independent influence factor affecting the morphology type of different calcification subtypes in PTC . The CVPTC group is mainly microcalcification ,and the TCV group is mainly macrocalcification .

Objective: To investigate the situation of hip dislocation with the application of "femur first" principle and "combined anteversion technique" during total hip arthroplasty. Methods: A retrospective analysis has been done on the clinical data about 104 patients(133 hips)who were diagnosed as hip disease and were treated with total hip arthroplasty by the doctors from the Department of Joint Orthorpaedics of Shandong Provincial Hospital Affiliated to Shandong University from June 2014 to June 2016, and all the prostheses applied in the operation were cementless ones.Among them, 65 patients were males, 39 females and their age was 46.6 years (ranging from 23 to 76 years) .And 29 of them underwent bilateral hip operations and 75 unilateral ones.Seventy-six cases of aseptic necrosis of the femoral head in the terminal stage, 28 cases of hip dysplasia and osteoarthritis.Surgical approach: of all the operations, 103 hips were operated on with hardinge approach, 30 with posterolateral approach.During the operation, first of all, the femoral medullary cavity was broached and then the anteversion of intramedullary broacher was measured.After that, the anteversion of the acetabular cup was calculated as 37° minus the anteversion of the broacher, and the acetabular cup was implanted at that angle.The patients′ prosthesis combined anteversion, range of motion of the hip joint, operation time, hemorrhage amount, and complications had been kept record.One, three, and six months respectively after the operation, all the patients received outpatient review, and took anteroposterior and lateral position X-ray examination.Harris hip score had been applied to evaluate their hip function before the operation and six months after the operation. Results: All the patients had been operated on smoothly, with the operation time of(57.6±14.5)minutes(36-115 minutes)and hemorrhage amount of (336.5±50.8)ml(180-620 ml). The operation finding showed that the combined anteversion by employing "femur first" principle and combined anteversion techniqueis was (36.6±6.8)°. Also, range of motion of the patients′ hip conforms to the D′Lima criterion and no patients had experienced dislocation and some other serious complications.The follow-up ended at August 2017, which lasted for 12.2 months(6-18 months), and 88 cases(119 hips) were interviewed. The Harris hip score for the affected sides of the hip joints 6 months after the surgery was 94.6(86-100). Compared with the score of 29.8(12-43) before surgery, the t-test had proven its statistical significance(t=12.82, P=0.003). Conclusion Based upon the "femoral first" principle and "combined anteversion technique" during total hip arthroplasty, these operations has been carried out well enough to meet the patients′ needs of performing normal daily activities.

Objective:To investigate the relationship of HRAS gene mutation and lymph node metastasis and ultrasonographic characteristics in differentiated thyroid carcinoma (DTC).Methods:The clinical data of 162 patients with DTCs who underwent thyroidectomy and confirmed by postoperative pathological examination in Beijing Friendship Hospital from January 2014 to February 2019 were retrospectively analyzed. There were 139 patients with papillary thyroid carcinoma (PTC group) and 23 patients with follicular thyroid carcinoma (FTC group); the PTC were further classified as classic variant of papillary thyroid carcinoma (CVPTC, n=34), follicular variant of papillary thyroid carcinoma (FVPTC, n=36) and tall cell variant (TCV, n=69). Tissue HRAS mutation frequency was detected in 162 DTC patients and 19 patients with follicular adenoma (FA); blood HRAS mutation frequency was detected in 195 healthy subjects. The correlation between HRAS mutation (IVS1-82del gctgggcctggg) and cervical lymph node metastasis was analyzed, and the ultrasonographic characteristics of DTC patients were also analyzed. Results:The frequency of HRAS mutation in DTC patients was higher than that in healthy controls[37.0%(60/162) vs. 26.2%(51/195), χ2=4.538, P=0.03], while there was no significant difference between FTC and FA [39.1%(9/23) vs. 5/19, χ2=0.769, P=0.38]. In DTC patients there was no significant difference in cervical lymph node metastasis between HRAS mutation group and wild type group [57.3% (43/75) vs. 42.6% (32/75), χ2=1.898, P=0.16]. Among CVPTC, FVPTC and TCV patients, the rates of cervical lymph node metastasis were 7/12, 8/14 and 48.0% (12/25) in HRAS mutation group, while those were 50.0% (11/22), 40.9% (9/22) and 43.1% (19/44) in wild-type group, respectively (χ2=1.009, P=0.98).There were 9 patients with HRAS mutation in FTC group, and the cervical lymph node metastasis in mutation group and wild-type group was 5/9 and 4/14, respectively ( P=0.38). The ultrasonographic characteristics of PTC patients with HRAS mutation were more likely to have clear boundaries [66.7%(34/51) vs. 42.0%(37/88); χ 2=7.833, P<0.01] and not close to the membrane[84.3%(43/51) vs. 65.9%(58/88);χ2=5.506, P=0.02]. Conclusion:DTC patients are more likely to have HRAS (IVS1-82del gctgggcctggg) mutation, and the ultrasonic characteristics of DTC patients with HRAS mutation are likely to have clear boundaries and be not close to the membrane.