Aim To amplify P30 gene and express P30 fusion with GST Methods P30 gene was smplified from T. gondii chromosomal DNA and ligated to pGEM-T and pGEX-4T-1. Screening-positive recombinants were induced for expression, which was subsequently detected by WB Results P30 gene was amplified and GST-fusion was confirmed by rabbit antiT. gondii serum. Conclusions The construction of pGEM-T-P30 and pGEX-4T-1-P30, together with the recombinant protein would lay a base for further investigation of P30 at a molecule-level and application to diagnosis and vaccination

Adult ; Breast Neoplasms ; metabolism ; pathology ; secondary ; Carcinoid Tumor ; metabolism ; pathology ; surgery ; Carcinoma, Adenoid Cystic ; pathology ; Carcinoma, Lobular ; metabolism ; pathology ; secondary ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; surgery ; Chromogranin A ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Hysterectomy ; Keratins ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Ovarian Neoplasms ; metabolism ; pathology ; Sex Cord-Gonadal Stromal Tumors ; metabolism ; pathology ; Synaptophysin ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology ; surgery

Cochlear Implantation ; Cochlear Implants ; Guidelines as Topic

Objective To investigate the efficacy of medial plantar flaps with cutaneous nerves on fixing palm skin defects for patients injured while working.Methods All cases with palm skin defects who received medial plantar flaps grafting from June 2003 to January 2013,were analyzed retrospectively.The data included 13 males and 10 females,with the mean age of 36 years old (range,18-61 years old).There were 16 cases with right hand injury and 7 cases with left,whose skin defects ranged from 3.6 cm×2.8 cm to 7.0 cru×5.5 cm.All cases had skin and soft tissue injury while 5 cases with vascular and nerve injury,3 cases with tendon involvement and being exposed,6 cases with open metacarpal fractures.15 patients were treated with emergency surgery within 8 hours after injury,while 8 patients were treated 10 days after debridement surgery.Results Altogether,23 patients who received medial plantar flaps grafting operations,with palm skin defects,were included in this study.All cases were followed up for 3 to 36 months with a average period of 18 month.The grafted flaps range from 4.6 cm×3.8 cm to 8.0 cm×6.5 cm.Among the patients,19 of them recovered excellently with healing by first intention,with 7-14 d healing time,fully-retained hand functions and palm appearance,and 4-8 mm for between two points distinguish test.Flaps survived fairly well in 3 patients because partial necrosis happened at plantar skin graft donor site.After being debrided and dressed,the wound healed at second phase with a healing time of 12-21 d and 5-9 mm for between two points distinguish test.The operated hands could flex and extend functionally in the follow-up period.Only one patient did not recovered from medial plantar flap grafting and the flap did not survive operationally.The patient recovered after another operation with groin skin grafting later.According to Gu Yudong et al' evaluation criteria,such as improved hand function,19 cases were scored as excellent recovery,3 cases as good,and 1 as poor.The excellent rate was 95.7％ (22/23).Conclusion Medial plantar flaps with cutaneous nerves are ideal flaps for palm skin defects restoration,which characterized by the advantages of fixed anatomical position,high survival rate and being good to the functional recovery and palm appearance.

A novel drug delivery system combining oral fast dissolving film with sodium deoxycholate/phospholipid mixed micelles was prepared to increase the absorption of cucurbitacin B that is a poor aqueous solubility substance. Encapsulation efficiency, particle size, zeta potential, polydispersity coefficient, investigated the morphology, disintegration time of oral fast dissolving film and the pharmacodynamic properties of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles before and after solidified in mice were evaluated and compared. The oral fast dissolving film prepared in this study showed a homogeneous pale yellow and could completely disintegrated in the 30 s. It could meet the requirements of rapidly disintegrating fully. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles loaded in oral fast dissolving film were (43.36 +/- 2.12)%, (108.82 +/- 5.2) nm, (-34.18 +/- 1.07) mV, 0.088 +/- 0.012, respectively. The encapsulation efficiency, particle size, zeta potential, polydispersity coefficient of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles in solution were (41.26 +/- 2.22)%, (181.82 +/- 4.48) nm, (-30.67 +/- 0.81) mV, 0.092 +/- 0.012, respectively. The difference of pharmacodynamics among film of cucurbitacin B-loaded micelles, cucurbitacin B-loaded micelles and free cucurbitacin B in vivo was compared. Solubility of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles has also been greatly improved. The tumor inhibition rate of cucurbitacin B loaded in sodium deoxycholate/phospholipid-mixed micelles was significantly improved and did not change significantly before and after solidified. These showed that the sodium deoxycholate/phospholipid-mixed micelles could enhance the antitumor activities of cucurbitacin B and the stability of cucurbitacin B sodium deoxycholate/phospholipid-mixed micelles was improved significantly after solidified by oral fast dissolving film technology without pharmacodynamic properties changed significantly.
Animals ; Antineoplastic Agents ; administration & dosage ; chemistry ; Cell Line, Tumor ; Deoxycholic Acid ; chemistry ; Drug Carriers ; chemistry ; Humans ; Male ; Mice ; Neoplasms ; drug therapy ; Phospholipids ; chemistry ; Solubility ; Triterpenes ; administration & dosage ; chemistry

Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.

Wnt signaling has been shown to inhibit adipogenic differentiation while inducing the osteogenic pathway of bone marrow stromal cells (BMSC). Patients with aplastic anemia (AA) often show excess fat accumulation in the bone marrow, possibly due to overactivation of the adipogenic pathway. Therefore, an activator of the Wnt signaling may alleviate the symptoms by enhancing the inhibition on the differentiation of BMSC towards adipocytes. To judge this hypothesis, the therapeutic effects of Wnt signaling activator lithium chloride (LiCl) combined with the currently used immunosuppressor cyclosporine A (CsA) on mice with AA in vivo was investigated. Mouse model with AA was established and the disease was confirmed by increased fat cell counts and decreased hematopoietic cell counts in the bone marrow of these animals. These mice treated with CsA 50 mg/(kg·d) alone or together with LiCl 20 mg/(kg·d), once daily for 5 d, then at day 14, 21 and 28 after establishment of mouse model with AA, the treatment effects were observed, including peripheral blood cells, bone marrow mononuclear cells (BMMNC) count and bone marrow biopsy examination in each group. The results showed that compared with the AA group, Hb content, WBC and BMMNC counts of CsA group and the combination group significantly increased. HE staining of bone marrow biopsy sample showed that the fat cells were significantly reduced in the bone marrow cavity (P < 0.05). Compared with the CsA group, Hb content, WBC and BMMNC counts of the combination group significantly increased (P < 0.05); HE staining of bone marrow biopsy sample showed that fat cells were reduced, the hematopoiesis of bone marrow was close to the normal. It is concluded that Wnt signal activator (LiCl) combined with CsA displayed a better treatment effect on AA in mouse models than the effect of using CsA only. They can promote the hematopoietic function of bone marrow, which may correlate with inhibiting differentiation of bone marrow mesenchymal stem cells into the fat cells by Wnt signaling.
Anemia, Aplastic ; drug therapy ; metabolism ; Animals ; Bone Marrow Examination ; Cyclosporine ; therapeutic use ; Disease Models, Animal ; Drug Therapy, Combination ; Female ; Lithium Chloride ; therapeutic use ; Mice ; Mice, Inbred BALB C ; Wnt Signaling Pathway

Objective To investigate the influence of nerve growth factor (NGF) on the ability of differentiation and proliferation of regulatory T cells (Tregs) induced by mesenchymal stem cells (MSCs).Methods The MSCs were stimulated directly by NGF.IL-10,TGF-β and HLA-G were tested.The expression of CD4 and CD25 was detected by flow cytometry after co-culture.The expression of CD4,CD25 and Foxp3 was detected by flow cytometry after Transwell co-culture.Results As compared with control group,the expression of IL-10,TGF-β and HLA-G in NGF group was increased (P＜0.05 for all).The number of Tregs was increased after the co-culture (P＜0.05).The reduction in IL-10 and TGF-β could block the inducing function of NGF (P＜0.05).Conclusion NGF can enhance the ability of differentiation and proliferation of Tregs induced by MSC,which is possibly associated with the increases in the expression of IL-10 and TGF-β.

OBJECTIVETo evaluate the value of the clinical use of CT perfusion imaging (CTPI) and CT subtraction angiography (CTSA) for diagnosing acute ischemic cerebrovascular disease (AICVD).

METHODSTwenty-four patients with AICVD onset within 24 hours were examined with regular CT, CTPI, and CTSA. Some cases received CTPI, magnetic resonance imaging (MRI), magnetic resonance angiography (MRA), digital subtraction angiography (DSA) or single photon emission computer tomography (SPECT) during follow-up examinations.

RESULTSOf the 24 cases, 11 had negative results from regular CT scans 3 - 6 hours after onset of stroke in 6 cases, 6 - 12 hours in 3 cases, and 12 - 24 hours in 2 cases. Ten of these cases were then confirmed by CTPI as having ischemic lesions, 2 with middle cerebral artery occlusion (MCAO), and 1 case with transient ischemic attack (TIA) with CTPI negative. Of the 24 cases, 13 had positive results from regular CT, 9 were diagnosed with ischemic lesions larger by using CTPI than regular CT, 1 case had MCAO and 1 had internal carotid artery occlusion (ICAO). There were 4 cases with ischemic lesions observed with regular CT having nearly the same range as that of lacunar infarctions using CTPI. Another 4 cases had more than 2 lesion areas. The peak time (PT), mean transit time (MTT) and relative flow (RF) of 24 cases were markedly different. The sides of ischemic lesions compared to each other and the core of the lesion compared to peripheral zones were also altered significantly (P < 0.01).

CONCLUSIONSCombined CTPI with CTSA can detect acute ischemic lesions at early and hyper-early stages and could distinguish between TIA, lacunar infarction and a larger area of infarction. Using semiquantitative blood perfusion analysis status, CTPI with CTSA could define position, area and range of the ischemic lesion and penumbra. These scans can also analyze the brain blood perfusion status. It is important to early diagnose the occlusion of the entire division of the internal carotid artery or middle cerebral artery and it is meaningful to assess prognosis and assignment of therapy.

Adult ; Aged ; Angiography, Digital Subtraction ; Brain Ischemia ; diagnostic imaging ; Cerebral Angiography ; Cerebrovascular Circulation ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

Objective To observe the effects of different energy of extracorporeal shock waves (ECSWs) on diabetic neuralgia in rats.Methods Fifty clean-grade healthy male Sprague-Dawley rats of both sexes,aged 8 weeks,weighing 180-200 g,were divided into 5 groups (n=10 each) using a random number table method:control group (group C),diabetic neuralgia group (group DN),low-energy ECSW group (group L + DN),medium-energy ECSW group (group M + DN),and high-energy ECSW group (group H+DN).Diabetic neuralgia models were established by intraperitoneally injecting streptozotocin (60 mg/kg) in DN,L+DN,M+DN and H+DN groups.ECSWs at 1,2 and 3 bar were applied during 4 consecutive weeks after successful establishment of the model once a week (T1-T4) in L+DN,M+DN and H+ DN groups,respectively.The mechanical paw withdrawal threshold (MWT),thermal paw withdrawal latency (TWL) and motor nerve conduction velocity (MNCV) were measured at T1-T4.Animals were sacrificed after the last measurement,and the sciatic nerve samples were obtained for determination of the expression of tumor necrosis factor-alpha (TNF-α) and interluekin-6 (IL-6) (by Western blot) and expression of TNF-α and IL-6 mRNA (by real-time polymerase chain reaction).Results Compared with group C,MWT,TWL and MNCV were significantly decreased at T1-T4,and the expression of TNF-α and IL-6 protein and mRNA was up-regulated in the other groups (P＜0.05).Compared with group DN,MWT at T2-4 and TWL and MNCV at T3,4 were significantly increased,and the expression of TNF-α and IL-6 protein and mRNA was down-regulated in L+DN,M+DN and H+DN groups (P＜0.05).Compared with group H+ DN,MWT at T2-4 and TWL and MNCV at T3,4 were significantly increased,and the expression of TNF-α and IL-6 protein and mRNA was down-regulated in L+DN and M+DN groups,and the expression of IL-6 mRNA was significantly down-regulated in group L+DN (P＜0.05).Conclusion ECSWs can mitigate diabetic neuralgia in rats,and the low-and medium-energy ECSWs produce better efficacy,and the mechanism is related to inhibiting inflammatory responses.