Objective:To observe the effects of Etifoxine on proliferation and migration of RSC96 (Schwann cells of rat) and its potential molecular mechanisms.Methods:From March, 2020 to October, 2020, cultured RSC96 were treated with 20 μmol/L Etifoxine and saline respectively for 48 h. Cell proliferation was tested by EdU assay using Cell-Light EdU DNA Cell Proliferation Kit and the capability of migration was determined by wound healing assay and a transwell system. To investigate the effects of Etifoxine on CELSR2 protein expression, after treated with different concentrations of Etifoxine at 0-20 μmol/L for 48 hours, cells were subject to Western blot analysis to verify the expression of CELSR2 protein. To explore whether CELSR2 would be a potential target of Etifoxine, siRNA targeting CELSR2 and control siRNA groups were transfected into 20 μmol/L Etifoxine-treated RSC96 using Lipo3000. Again, the cell proliferation and migration of were investigated after 48 hours with the same procedures. The two-tailed Mann-Whitney U test was employed in statistical assessment.Results:EdU results showed a significant higher percentage of Edu-positive (proliferating) cells in the 20 μmol/L Etifoxine-treated group than the control group[(36.30±3.09)% vs (19.40±2.50)%, P<0.05]. Transwell migration assay demonstrated that the number of 20 μmol/L Etifoxine-treated RSC96 which migrated through the transwell membrane was higher than saline group, with significant statistical difference [(132.30±6.77) vs(65.33±7.37), P<0.05]. The percentage of reduction of wound area measured at 24 hours and 36 hours after the scratch also showed the similar results [(30.67±2.16)% vs (23.00±2.61)%; (86.00±2.19)% vs (49.67±2.81)%, respectively, P<0.05]. Besides, with increase of the concentration of etifoxine, the expression of CELSR2 showed an trend of increase in RSC96 ( P<0.05), but no significant statistical difference was found between 10 μmol/L and 20 μmol/L groups ( P>0.05). Interestingly, the rate of cell proliferation, the number of migrating cells and the percentage of wound area reduction of RSC96 in which were treated by Etifoxine and transfected with CELSR2 siRNA were significantly decreased compared with the control siRNA treatment ( P<0.05). Conclusion:Etifoxine could promote proliferation and migration of RSC96. Upregulation of CELSR2 protein expression in RSC96 is associated with the Etifoxine-induced enhancement of cell proliferation and migration.

Objective To prepare a peptide monoclonal antibody (McAb)against human papillomavirus 18E6 separately,and identify its specificity and pathogenicity.Metheds The advantage epitope peptide was designed and synthesized by ABCpred and Bcepred,and then used to immunize BALB/c mice after coupling with bovine serum albumin (BSA).And the McAb was prepared by hybridoma technique.HPV18E6 gene was amplified from cervical swab specimen containing HPV18 and insert-ed into expression vector pET-28a.The constructed recombinant plasmid was transformed to E.coli BL21(DE3)for expres-sion under induction of isopropyl thio-β-D-galactoside.The expressed protein was used to identified the McAb had been pre-pared.Results The hybridoma cell lines could constantly produce MAbs against HPV18E6 peptides.Sequencing proved that recombinant plasmid pET-28a-HPV18E6 was constructed correctly.Western blotting showed that the anti-HPV18E6 pep-tides antibody could specifically recognize HPV18E6.Conclusion A monoclonal antibody against the advantage epitope pep-tide of human papillomavirus 18E6 prepared could specifically recognize HPV18E6 specifically.

BACKGROUND: Compared with single proprioceptive training and simple neuromuscular training in the trunk and hip, the integrative neuromuscular training possesses comprehensive effect, which not only prevents sport-induced knee injury, but also improves the athletes' sport ability.OBJECTIVE: To validate whether the integrative neuromuscular training can enhance sport quality of juvenile female athletes and improve biomechanical pattern of the lower limb when jumping.METHODS: Eighteen juvenile female tennis athletes were selected and randomized into experimental and control groups. The athletes in the experimental group received 8-week integrative neuromuscular training and technical training involving the strength of the lower limb, core stability, plyometrice training, dynamic equilibrium, agility and speed. Those controls were subjected to 8-week traditional physical training and technical training. The vertical jump dynamic equilibrium, agility and biomechanics of the lower limb were measured at 1 week before intervention and 7 weeks after intervention to quantize the effect of integrative neuromuscular training on the sport ability and prevention of knee injury.RESULTS AND CONCLUSION: The vertical jump, agility, overall stability index, the knee extension angle as well as knee adduction and abduction moment in the experimental group were significantly improved (P < 0.05). Integrative neuromuscular training cannot only enhance the explosive strength of the lower limb, dynamic equilibrium, and agility when juvenile female athletes exercising, but also improve the biomechanics of the lower limb when jumping, increase the knee extension angle, and reduce knee adduction and abduction moment

We reported a 23-year-old man who was involved in a high-speed motorcycle accident. He sustained a closed fracture at the right distal femur. The primary fracture happened on February 2008. He underwent open reduction and internal fixation with cloverleaf plate. And one hundred days after the surgery, the proximal screws were pulled-out, but the bone union was not achieved. Treatment consisted of exchanging the cloverleaf plate with a locking compression plate and using an auto-iliac bone graft to fill the nonunion gap. In July 2009, the patient had a sharp pain in the right lower limb. The X-ray revealed that the plate implanted last year was broken, causing a nonunion at the fracture site. Immediately the plate and screws were removed and an intramedullary nail was inserted reversely from the distal femur as well as a 7 cm long bone from the right fibula was extracted and longitudinally split into two pieces to construct cortical bone plates. Then we placed them laterally and medially to fracture site, drilled two holes respectively, and fastened them with suture. We carried on auto-iliac bone grafting with the nonunion bone grafts. The follow-up at 15 months after operation showed that the treatment was successful, X-ray confirmed that there was no rotation and no angular or short deformity. We briefly reviewed the literature regarding such an unusual presentation and discussed in details the possible etiology and the advantages of autologous double-strut fibular grafts to cope with such an intractable situation.
Bone Plates ; Femur ; Fibula ; Fracture Fixation, Internal ; Fractures, Bone ; Humans

OBJECTIVETo study the effect of dihydroartemisinin (DHA) combined irradiation on the apoptosis of human lung cancer GLC-82 cells and to study its mechanism.

METHODSThe growth inhibition rate of GLC-82 cells acted by different concentrations DHA was detected using MTT assay at 24, 48, and 72 h, respectively. Clone forming test was used. With multi-target single-hit model, the radiosensitization effect was assessed by calculating sensitizing enhancement ratio (SER).The effect of DHA combined irradiation on the apoptosis of GLC-82 cell cycle distribution and apoptosis were measured by flow cytometry. The protein expression of p53, p21, Bcl-2, and Bax were detected by Western blot.

RESULTSDifferent concentrations DHA (4, 8, 16, 32, 64, and 128 μg/mL) had cytotoxicity on GLC-82 cells. The IC50 for 24, 48, and 72 h was 38.25,20.58, and 10.36 μg/mL, respectively, in obvious dose- and time-dependent manner. The growth inhibition rate was more significantly increased than that of the blank control group (P < 0.01, P<0.05). DHA had sensitization enhancement effect on GLC-82 cells, with SER of 1.4. DHA combined irradiation could obviously change the structure of GLC-82 cells cell cycle and induce apoptosis (with the apoptosis rate of 21.5%), which was significantly different from that of the blank control group (P < 0.05). Western blot showed the expression of p53 and p21 protein could be increased by DHA combined irradiation, and the expression of Bcl-2 protein down-regulated (P <0.01, P <0. 05).

CONCLUSIONSDHA had stronger cytotoxicity and radiosensitization on GLC-82 cells. Its mechanisms might lie in making the arrest of GLC-82 cells' growth at G0/G1 phase, decreasing the ratio of cells at S phase, restoring the function of p53, decreasing the expression of Bcl-2 protein, and inducing apoptosis in GLC-82 cells.

Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Flow Cytometry ; Humans ; Lung Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; Radiation-Sensitizing Agents ; pharmacology ; Tumor Cells, Cultured ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo evaluate the feasibility and safety of nickel-titanium compression anastomosis ring (CAR27) in colorectal anastomosis after low anterior rectal resection in animal models.

METHODSEnd-to-end colorectal anastomosis was performed using CAR27 in 6 experimental pigs after resection of the middle and lower third of the rectum. The animals were observed postoperatively for up to 56 days. Five pigs were sacrificed at day 14 and the other at day 56. Distance from anal verge to anastomosis and anastomotic circumference were measured. Histopathologic examination was performed.

RESULTSThe median distance from anal verge was 5.3(4-6) cm. No anastomotic leak or other complications were observed. All the pigs recovered and gained weight. In 5 animals sacrificed at day 14, the mean circumference of the anastomosis was 6.8(6.5-7.0) cm, and histopathological examination showed mild inflammatory reaction and fibrosis. In the one sacrificed at day 56, the circumference expanded to 9.3 cm, and no inflammation and fibrosis were observed. Minor adhesion was noticed in only one pig, while smooth and intact serosa in the anastomosis was seen in the rest of the animals.

CONCLUSIONCAR27 is a promising device for mid and low colorectal anastomosis.

Anastomosis, Surgical ; instrumentation ; Animals ; Female ; Male ; Models, Animal ; Nickel ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Swine ; Swine, Miniature ; Titanium

OBJECTIVETo study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).

METHODSThe effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.

RESULTS(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.

CONCLUSIONMSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogenes ; Receptor Protein-Tyrosine Kinases ; metabolism

Sorafenib is a multi -targeted tyrosinekinase inhibitor , and mainly used for the treatment of advanced renal cell carcinoma and hepa -tocellular carcinoma.The pharmacokinetic changes of sorafenib caused by drug -drug interactions may affect its clinical efficacy and safety, so individualized treatment drug monitoring could control the occurrence of potential drug interactions.In this paper, the clinical applications and drug -drug interaction of sorafenib was reviewed .

Objective To explore the surgical skills and efficacy of microsurgical excision of large acoustic neurinoma via the suboccipital retrosigmoid keyhole approach. Methods Fifty-nine patients with large acoustic neurinoma (≥3 cm) underwent microsurgical resection via suboccipital retrosigrmoid keyhole approach. The intraoperative position of the operative bed and angle of the microscope were adjusted to expose the tumors sufficiently. The duramater was sutured tightly and the bone flap was replaced and fixed. Results Of the 59 patients, 53 (89.8%) received a total resection of the tumors, and 6 (10.2%) subtotal resection. No patient died. The facial nerve was anatomically preserved in all the patients. Forty-five patients (76.3%) suffered from mild or moderate facial palsy after the surgery, and the symptoms were improved significantly after expectant treatment. Fifteen patients preserved partial hearing. No subcutaneous water accumulation or cerebrospinal fluid leakage occurred.Conclusion Microsurgery via suboccipital retrosigmoid keyhole approach is a favorable treatment for large acoustic neurinomas with low morbidity and mortality,which can effectively protect the function of the acoustic and facial nerves.

BACKGROUNDIt remains a challenge to inhibit the local recurrence or distant metastasis of localized or locally advanced renal cell carcinoma (RCC) after surgical resection. We investigated the feasibility, safety and efficacy of immunotherapy using autologous tumor lysate (TL)-pulsed dendritic cells (DCs) and cytokine-induced killer (CIK) cells in patients with localized or locally advanced RCC.

METHODSFrom January 2001 to July 2009, we collected 137 patients that met the selection criteria and randomly divided them into three groups. After surgery, immunotherapy with TL-pulsed DCs-CIK cells (DC-CIK group) and interferon (IFN)-α (IFN-α group) was performed in 46 patients, respectively. The other 45 patients received no postoperative adjuvant therapy (the control group). The changes in the numbers of T lymphocyte subsets, including CD4(+)CD25(high) regulatory T cells (Treg), were determined before the operation and after immunotherapy. The overall survival was compared among the three groups.

RESULTSAn increase of the CD4(+)/CD8(+) ratio and a decrease of CD4(+)CD25(high) cells were observed after TL-pulsed DC-CIK cells or IFN-a immunotherapy. All patients tolerated the TL-pulsed DC-CIK cells immunotherapy very well, and side effects in the DC-CIK group were less than in the IFN-α group. The metastasis and recurrence rates were significantly decreased after TL-pulsed DC-CIK cells or IFN-α immunotherapy compared with the control group (P < 0.01). The Log-rank test showed that the overall survival rates were significantly higher in the DC-CIK group and IFN-α group than that in the control group (P < 0.01), but there was no difference between the DC-CIK group and IFN-α group (P > 0.05).

CONCLUSIONPostoperative immunotherapy with TL-pulsed DC-CIK cells may prevent recurrence/metastasis and increase the overall survival rate after surgery in localized or locally advanced RCC.

Adult ; Aged ; Carcinoma, Renal Cell ; immunology ; mortality ; therapy ; Cytokine-Induced Killer Cells ; immunology ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy ; Kidney Neoplasms ; immunology ; mortality ; therapy ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology