Objective: Proto-oncogene Zbtb7A has been characterized as a molecular switch in the process of cancer initiation and development.Our goal is to obtain the POZ domain of the Zbtb7A protein and prepare its polyclonal antibodies.Methods: We optimized the coding sequence of the POZ domain according to the codon bias of E.coli and synthesized the sequence with two-step PCR,which was then introduced into the pET-26b(+) vector to express the protein.The recombinant protein was analyzed by 15% SDS-PAGE and the corresponding band was cut out from gel.The minced gel slice that contained the POZ domain protein was injected to immunize rabbits.The collected rabbit antiserum was purified using the saturated ammonium sulfate method in combination with protein G antibody purification,and the purified polyclonal antibodies was evaluated by Western blot.Results: The optimized sequence of the POZ domain was correctly obtained and successfully constructed into the pET-26b(+) vector.After induction,an expected protein band about 14 KD was detected on 15% SDS-PAGE,and highly purified polyclonal antibodies were obtained,which were specifically bound to human Zbtb7A.Conclusion: The obtained recombinant protein of the POZ domain and its polyclonal antibodies can be used for further studies of Zbtb7A.

Objective This study is to identify long-term functional recovery and maturity of regenerated nerve fibers after repairing mouse nerve defects with chitosan/polylactide-co-polyglycolide artificial nerve grafts ( CPANGs ) . Methods Mouse sciatic nerve defects, 2mm in length, were bridged by CPANGs (n=6), with nerve autograft (n=6) and nerve defect (n=6) as controls.Plantar test, electrophysiological examination and laser Doppler perfusion imaging following nerve crush were carried out at 1 year after repair to assess nerve function recovery , while muscle wet weight ratio, histological assessment and transmission electron microscopy were performed to evaluate nerve re -innervation and maturity of regenerated nerve fibers .Results When compared to the autograft group , the CPANG group did not show statistically significant difference in functional recovery in terms of paw withdrawal latency , neurogenic vasodilatation , amplitude and latency of compound muscle action potentials ( CMAPs ) , wet weight ratio of gastrocnemius and tibialis cranialis muscles , number of myelinated nerve fibers and density of unmyelinated axons .However , both these two repair groups exhibited significantly longer CMAPs latency , thinner myelin sheath and a lag-behind shift of diameter distribution of myelinated axons as compared to the normal control .Conclusion At 1 year after the mouse sciatic nerve defect was repaired by CPANGs , sensory and autonomic nerve function , number of regenerated axons and muscle re-innervation degree were recovered to the same extent as nerve autografting , but the regenerated nerve fibers were in a state of immaturity .

Objective: To determine the role of cross-talk between calcineurin-dependent signal transduction pathway and protein kinase C (PKC), mitogen-activated protein kinase (MAPK) and protein kinase A (PKA) in airway remodeling in asthma. Methods: Male guinea pigs were sensitized with intraperitoneal injections of ovabumin (OVA), then treated with cyclosporin A (CsA,5 mg/kg), an inhibitor of calcineurin, then inhaled OVA for 2 weeks 14 days later. Activities of calcineurin, PKC, MAPK, and PKA were was analyzed by phosphorylation and dephosphorylation. In primary cultures of rat airway smooth muscle cells (ASMC), activities of calcineurin, PKC, MAPK, and cross-talk induced by urotensin Ⅱ (UⅡ), a recently identified strong mitogen, were measured. Results: (1) The activities of calcineurin, MAPK and PKC increased by 19% (P0.05). (4) CsA 10 -6 mol/L inhibited UⅡ-stimulated PKC activity by 14% (P0.05). Conclusion:The signal transduction pathways between calcineurin and other protein kinases such as PKC, MAPK and PKA have cross-talk in airway remodeling in asthma.

Abstract: Objective　To report a case of suppurative knee arthritis caused by Pasteurella multocida and review relevant literature to improve the awareness of the clinical physicians regarding this bacterium and provide reference for clinical diagnosis and treatment. Methods　A case of right knee suppurative arthritis caused by Pasteurella multocida was retrospectively reported and relevant literatures were reviewed in this article. Results　The infected person was a 76-year-old female patient with a 5-year history of intermittent pain in his right knee and suffered from joint swelling, aggravation pain, and limited flexion and extension activities after intraarticular injection of sodium hyaluronate. After admission and completing all necessary tests, the patient was later confirmed to have been infected with Pasteurella multocida. The patient's right knee was promptly examined and cleared under arthroscopic surgery, synovium and meniscus were excised, a drainage tube was inserted, and continuous joint cavity irrigation was performed after the surgery, and then ceftriaxone was injected and amoxicillin/clavulanate potassium was taken orally for anti-infection and the patient's condition improved significantly after 26 days. Conclusions　Pasteurella multocida infection cases are relatively rare, but the consequences in high-risk groups are relatively serious. Therefore, awareness of Pasteurella multocida and infection caused by it should be improved and high-risk groups should try to avoid contact with infectious sources as well as strengthen the management of pets so as to avoid infection.

Objective To investigate the effect of Ligu Capsules in preventing and treating menopausal osteoporosis in ovariectomized rats.Methods Rat models with osteoporosis were established by ovariectomy. Bone mineral density of femur,dry and wet bone weight and blood biochemical parameters were measured before and after treatment.Results Compared with the model group, Ligu Capsules increased femur bone mineral density,elevated the serum levels of Ca,P,Mg ,Zn and CT, decreased serum BGP level and hydroxyproline/creatinine(Hyp/Crea)ratio,increased dry weight, wet weight and ash weight of femur, as well as the bone length, bone volume, and bone diameter.Conclusion Ligu Capsules can prevent and treat osteoporosis in ovariectomized rats.

This study is to examine the effects of NNIspm-mediated cellular senescence of HepG2 cells and elucidate its potential molecular mechanism. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Cell cycle distribution, intracellular fluorescence intensity and accumulation of intracellular reactive oxygen species (ROS) were detected by high content screening (HCS). Protein expression was detected by Western blotting. Polyamines content was analyzed by high performance liquid chromatography (HPLC). The results demonstrated that NNIspm significantly induced HepG2 cells senescence. This effect was due to the decrease of intracellular polyamines, the arrest at G0/G1 phase and an increase of ROS level. The molecular senescence marker p21 increased significantly after NNIspm treatment. In contrast, the protein expressions of Cyclin E and CDK2 were obvious down-regulation. The results indicated that cellular senescence induced by NNIspm was one of its antitumor mechanisms.

Aim To investigate the apoptotic mechanism and polyamine transporter recognition of 3-nitro-naphthalimide norspermine conjugate (NNINspm),a novel naphthalimide-polyamine conjugate, in HepG2 cells.Methods The cytotoxicity of NNINspm was assessed by MTT assay.Cell cycle distribution and apoptosis were measured by flow cytometry.The protein expression of cytochrome C,14-3-3,Bad,Bcl-xL,mTOR,p70S6K,Cdk4,p27~(kip1),Akt,Caspase-3,Caspase-9 was evaluated by Western blot.The translocation of Akt was detected by high content screening (HCS) analysis.Results NNINspm induced HepG2 cells apoptosis via Akt dephosphorylation and then triggered a series of signal events, such as Bad dephosphorylation, dissociation of 14-3-3 and Bad, and then binding to Bcl-xL,which finally resulted in mitochondrial disruption,cytochrome c release and caspase cascade activation.Furthermore,the NNINspm-mediated cell cycle arrest was due to mTOR and p70S6K dephosphorylation,Cdk4 down-regulation and p27~(kip1) up-regulation.Conclusion NNINspm induces HepG2 cell apoptosis via PI_3K/Akt signal pathway.

Objective To study the diagnosis, treatment and prognosis of the rupture of breast implant. Methods Physical examination, X-ray fluoroscopy, high frequency and color the Doppler ultrasonography were helpful to make a clear diagnosis, in exploring and removing the fiber envelope by inframammary or periareolar incision, repairing the anatomy structure of the breasts and performing augmentation mammaplasty again. Results In 32 patients, the breast shape was basically symmetrical, most of the incisions were healed by first intention except 2 cases with complication of subcutaneous fluid collection. A satisfactory therapeautic result could be achieved by taking out the breast implant which was rupture, exploring and removing the fiber envelope, and extending the lacuna. Conclusion A satisfactory therapeautic result can be achieved by extending the lacuna.

In the present study, the apoptotic mechanism and polyamine transporter recognition of WJH-6, a novel polyamine conjugate, were investigated in K562 and HL-60 cells. The cytotoxicity of WJH-6 was assessed by MTT assay; cell cycle distribution and apoptosis were measured by flow cytometry; the protein expression of Caspase-3, Caspase-8, Caspase-9, Bid and mitochondrial membrane potential (MMP) were evaluated by high content screening (HCS) analysis; the protein expression of cytochrome c was measured by Western blotting. The results showed that WJH-6 could be recognized and transported by polyamine transporter (PAT). Furthermore, WJH-6 was able to inhibit K562 and HL-60 cells proliferation and induce apoptosis. This apoptotic effect was relative to MMP loss, cytochrome c release from mitochondria to cytoplasm and the activation of Caspase-8, Caspase-9, Caspase-3 and Bid. These results suggested that WJH-6-induced K562 and HL-60 cells apoptosis was related with mitochondrial damage.

Objective The objective of this study was to investigate the effect of acidic conditions on epithelial mesenchymal transition(EMT) of HepG2 cells.Methods HepG2 cells were cultured under acidic and alkaline conditions to observe the difference of cell morphology.Wound healing assays were performed to detect the migration ability of the two groups.Matrigel invasion assays were used to detect the invasive ability of the cells.The expression of EMT-related genes at mRNA level was detected by RT-PCR.The expression level of EMT-related gene protein was detected by Western blot.Results The morphology of HepG2 cells was changed from epithelium to interstitial type under acid conditions.The migration ability of HepG2 cells under acidic condition was stronger than that of alkaline conditions.The number of HepG2 cells crossing Matrigel was higher than that of alkaline condition.The mRNA expression of Vimentin,Slug,Snail and Zeb1 related to EMT and the protein expression of Vimentin and MMP9 related to EMT in HepG2 cells were significantly higher than those under alkaline condition.Conclusion The acid conditions can induce the occurrence of EMT in HepG2 cells.