Objective To construct a novel tissue?engineered bone matrix gelatin (BMG)/poly[butylene succinate?co?tere?phthalate] (PBST) biphasic scaffold for annulus fibrosus. Methods The PBST spinning fibers were prepared by electrospinning and the porosity and water absorption rate were tested. Rabbit annulus fibrosus cells were isolated, cultured and identified through SafraninOstaining, and collagenⅡimmunohistochemical staining in vitro. And then annulus fibrosus cells were implanted on the PBST fiber, whose growth situation was observed by scanning electron microscope (SEM). Then the BMG/PBST biphasic scaf?fold was constructed by BMG as the outer annular fibrosus and PBST fiber as the inner annular fibrosus. The annulus fibrosus cells were implanted on the biphasic scafflod and cultured for 3, 7 and 21 days in vitro. The biomechanical and biological property was observed at the predetermined time point. Results The porosity of the fiber was 61.83%±7.33%and its water absorption rate was 297.34%± 57.13%. The identified result of annulus fibrosus cells were positive, suggesting that the cells have still kept their annulus fibrosus cells characteristics. The cells growth could be observed through SEM at 3rd and 7th day after implanted on the fi?bers. After cultured on the BMG/PBST scaffold, HE staining proved that the cells could ingress into the inner of fiber with time. SafraninOstaining and collagenⅡimmunohistochemical staining proved that the cells can secreted abundant proteoglycan and collagenⅡ, the special annulus fibrosus cell extracellular matrix. Compared with the BMG/PBST scaffold without cells, the elastic modulus of biphasic scaffold was increased from 14.83±1.02 MPa to 17.56±1.47 MPa after cultured with cells for 21 days in vitro. Conclusion The novel tissue?engineered biphasic scaffold for annulus fibrosus constructed with BMG and PBST fiber spinning has good cytocompatibility and biomechanical characteristics, which provide a basis for the complete tissue engineered interverte?bral disc.

The purpose of this study was to evaluate the effect of adenosine A2A receptor antagonist ZM241385 on amygdala-kindled seizures and its roles in epileptogenesis. Electrodes were implanted into the right amygdala of male adult Wistar rats. Kindling was accomplished by using stimulus strength of 500 μA applied daily to the amygdala until 10 consecutive stage 5 seizues were induced. Then effect of ZM241385 was studied in fully kindled rats after intracerebroventricular administration of the drug. In addition, the effect on kindling progression was evaluated through ZM241385 injection before daily stimulation. In all experiments, behavioral changes in the rats in response to ZM241385 were monitored closely. The results showed that, in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) decreased afterdischage duration (ADD), motor seizure duration (MSD), stage 5 duration (S5D) and seizure duration (SD), but only the effect on ADD was dose-dependent. The doses of 0.001-0.1 nmol/L had no influence on stage 4 latency (S4L) and seizure stage (SS). The dosages of 0.0001 and 1 nmol/L of ZM241385 did not exert any effect on all seizure parameters. In contrast to the results in fully amygdala-kindled rats, ZM241385 (0.001-0.1 nmol/L) had minimal or no effects on the progression of amygdala-kindled seizures. We are led to the conclusion that although ZM241385 had no influence on the progression of amygdala-kindled seizures, it had potent anti-convulsant profile and little adverse effects at the dosage of 0.001-0.1 nmol/L, suggesting that the agent is effective against the amygdala-kindled seizures.

Objective To investigate the dynamic changes of adenosine receptors, A1 (A1R) and A2a (A2aR) in the brain from the acute to chronic phase after kindling and to explore the correlation between seizure and expression level of A1R and A2aR. MethodsRats were randomly selected into the testing model, reference and normal control groups.Testing rats were kindled by lithium choride-pilocarpine, reference rats were treated with saline, and no treatment was given in normal control group. The dynamic expression of A1R and A2aR were detected by RT-PCR, immunofluorescence staining and Western blot at time-points of 24 hours, 1 month and 6 months post-kindling. Results In the acute phase of 24 hours after kindling, the A1R expression level (mRNA level was (1. 1483 ±0. 1182); Western blot result was ( 0. 7872± 0. 0621 ) ; immunofluorescence staining count was ( 76. 17 ± 4. 62 )/HP) was increased and A2aR (mRNA level was (0. 8338±0. 0572) ; Western blot result was (0. 2098 ±0. 0257) ; immunofluorescence staining count was (43. 83 ± 5. 12 )/HP) was decreased.The results showed statistically difference compared with the reference and normal groups (P< 0. 05 ). In the later chronic phase of 1 month and 6 months after kindling, the expression level of A1R was decreased and A2aR was increased. These data revealed statistically significant difference (P <0. 01 ). Furthermore, the comparison of the results in 1 month and 6 months after kindling found that the expression of AIR was lower in 6 months (mRNA level was (0. 5682 ±0. 0443) ; Western blot result was (0. 7749 ±0. 0262) ; immunofluorescence staining count was (38. 50 ±4. 81 )/HP) than in 1 month and that of A2aR was higher in 6 months (mRNA level was (1. 2169±0. 0332) ; Western blot result was (0. 7080 ±0. 0371 ); immunofluorescence staining count was (114. 50 ± 4. 04)/HP). The differences were statistical significant (t = - 19. 02--13.28, P < 0. 05). ConclusionsThe expressions of A1R and A2AR during and after kindling presents a bidirectional change. In the acute phas the expression of AR is regulated to suppress seizures. While in the chronic phase, the repeated seizures result in the change of A1R and A2aR expression in the opposite direction. This mechanism plays an important role in refractory seizures.

Objective To assess the clinical value of laparoscopic colorectal resection combined with simultaneous radiofrequency ablation for liver metastases.Methods Twenty-two patients with colorectal cancer and synchronous liver metastases were treated by laparoscopie colorectal resection and simultaneous radiofrequency ablation(RFA)in Third People's Hospital of Chengdu from December 2001 to July 2006.Areas of colliquative necrosis were detected by contrast-enhanced computed tomography postoperatively.The outcomes were analyzed by chi-square test.Results Of all patients,8 had muhiple liver metastases,16 had complications.Thirty-one liver metastases were treated by RFA,and no complication oecurred.The mean postoperative hospital stay was(14±5)days,and there was no perioperative death.Five of the 22 patients were identified as incomplete ablation of the tumor,and the RFA was reperformed.The recurrence of liver metastases in the RFA necrotic zone was observed in 4 patients,and RFA was reperformed on 2 of them.Six patients died,and 2 of them died of liver metastases recurrence in the RFA necrotic zone.The recurrence rate of liver metastases and mortality was 18%(4/22)and 27%(6/22),respectively.The recurrence rate of patients with the diameter of the metastatic lession≥2.0 cm was significantly higher than those with the diameter of the metastatic lession＜2.0 cm(x2=5.867,P＜0.05).Conclusions Laparoseopie colorectal resection combined with simultaneous RFA provide a curative opportunity for colorectal cancer patients who are old and have multiple liver metastases,complications,poor tolerance of operation,and for patients whose tumors are difficult to resect.

Objective To clinically analyze the frequent stroke localizations of moyamoya disease in order to improve our cognition toward it and reduce missed diagnosis. Methods All 32 patients were prospectively analyzed over the past 10 years in our hospital. Results The ratio of female to male was 1.28 and their age of onset ranged from 7 to 47 years old. Mean age of 5 ischemic stroke patients (15.6%) was 24 and mean age of 21 haemorrhagic stroke patients (65.6%) was 33 while mean age of 6 mixed type stroke patients (18. 8% ) was 32. The frequent ischemic stroke localizations were frontoparietal lobe (60%),temporo-occipital lobe (20%), and periventricular zone (20%). The frequent haemorrhagic stroke iocalizations were periventricular zone (42.8%), ventricle (39.2%), temporo-occipital lobe (10. 8%) and subarachnoid space (7.2%). No cerebellum and brain stem stroke occurred. Conclusion Adult patients usually develop intracranial hemorrhage. Moyamoya disease should be considered severely when adult patients develop periventricular and ventricular intracranial hemorrhage, frontoparietal cerebral infarction or adolescent patients develop ischemic stroke, especially companied with epilepsy.

Objective To investigate the effects of testosterone (T) replacement therapy (TRT) on carotid artery intima-media thickness (IMT) in middle aged and elderly male patients.Methods A total of 80 middle-aged and elderly male patients with testosterone deficiency and increased carotid artery IMT were selected and randomly divided into two groups:the treatment group (n=38,treated with testosterone for 1 year) and the control group (n=42,without any treatment).The serum T level,IMT and prostate-specific antigen (PSA) before and after treatment were determined.The correlation between the testosterone level and carotid artery IMT was analyzed.Results There were no significant differences in the serum T level and IMT between the control group and the treatment group before treatment [(10.39 ± 1.44) nmol/L vs.(10.88 ± 1.87) nmol/L,(1.25 ±0.11) mm vs.(1.24±0.13) mm,t=1.32,-0.26,P=0.191,0.794].Compared with pretreatment,the serum T level was significantly increased and the IMT was significantly decreased in the treatment group afterTRT [(10.88±1.87) nmol/L vs.(22.83±1.56) nmol/L,(1.24±0.13) mmvs.(1.18±0.16) mm,t=-29.14,2.55,P=0.000,0.015],while no significant differences in the serum T level and IMT were found in the control group before and after treatment [(10.39± 1.44)nmol/L vs.(9.99±1.72) nmol/L,(1.25±0.11) mm vs.(1.27±0.11) mm,t=1.24,-1.00,P =0.219,0.323].Linear correlation analysis showed that the serum T level was negatively correlated with IMT (r-0.605,P=0.000) and multiple regression analysis showed that the T level was an independent factor for IMT.Conclusions Testosterone replacement therapy is an effective treatment to alleviate IMT in middle-aged and elderly male patients,which may play an important role in preventing cardiovascular diseases in middle-aged and elderly male patients.

Objective To fabricate a novel tissue-engineered cartilage with adipose-derived stem cells (ADSCs) seeded on the chitosan hydrogel scaffold to repair articular cartilage defect.Methods Adipose tissue and costal cartilage were harvested from New Zealand rabbits,and ADSCs in passage one and chondrocytes were obtained after the samples were digested and cultured in vitro.ADSCs were digested,suspended,seeded onto the sterile chitosan gel,and cultured in vitro for 1 week to fabricate the tissue-engineered cartilage.The defects were respectively filled with the tissue-engineered cartilage (composite group),chondrocyte suspension (cell group),chitosan gel (material group) and nothing at all (control group).At postoperative 12 weeks,cartilage repair was evaluated using the gross examination,histological staining,immunohistochemical staining and international cartilage repair society (ICRS) histological score.Results Effect of cartilage repair in composite group was significantly better compared to other groups.The regenerated tissue in composite group seemed tightly bound in normal tissue,with similar structure and extracellular matrix secretion.ICRS histological score in composite group was (13.89 ± 0.14) points,which differed significantly from (7.06 ± 0.19) points in control group,(7.14 ± 0.22) points in cell group and (7.46 ± 0.26) points in material group (P ＜0.01).Conclusion The tissue-engineered cartilage with ADSCs seeded onto the chitosan hydrogel is effective for repair of articular cartilage defect.

BACKGROUND: A study combined medical CT technology and digital three-dimensional (3D) interactive control system Mimics software and computer simulation software tools for 3D reconstruction and simulation reconstruction of the femur. Customized individualized femoral prosthesis has high matching degree with human body structure, and has high application value in clinical research. OBJECTIVE: To investigate the application value of 3D simulation of femoral prosthesis in artificial femoral prosthesis replacement. METHODS: Totally 82 cases, who received femoral prosthesis replacement from July 2014 to June 2016 in Taihe Hospital of Shiyan, were enrolled and randomly assigned to three groups: personalized group (n=28), cementless group (n=27) and cemented group (n=27). Different types of femoral prosthesis were designed in each group. 3D model reconstruction in femur was designed using medical CT technology combined with computer. Prosthesis implantation was simulated in computer, and the relevant information was recorded. Simulated operation was conducted on the mechanical experiment of femoral prosthesis in the biped and single foot loads so as to provide the most suitable operation plan. The possible risk was forecasted and assessed for preoperative preparation. RESULTS AND CONCLUSION: (1) In the simulation of mechanical experiment, under double foot and single foot loads, after prosthetic replacement, stress in the femur was significantly better in the personalized group than in the cementless group and cemented group (P <0.05). (2) After replacement, normal stress and shear stress were significantly better in the personalized group than in the cementless group and cemented group (P < 0.05). (3) The initial, horizontal and vertical micromovement was significantly better in the personalized group than in the cementless group and cemented group (P < 0.05). (4) In summary, the use of computer technology and 3D CT software can make individualized femoral prosthesis for patients. Compared with the cementless and cemented prostheses, personalized prosthesis has obvious advantages in stress distribution, interface stress and initial micromovement.

From the process of human immunodeficiency virus-1 (HIV-1) invading cells, the combination of gp120 and CD4 is the first step for HIV-1 to invade cells. Interfering with this process can prevent HIV from recognizing target cells and inhibit virus replication. Therefore, HIV-1 gp120 is an important part of the HIV-1 life cycle. Fostesavir, a phosphatate prodrug derived from the gp120 inhibitor BMS-626529 modified by the prodrug strategy, was approved for the treatment of adult patients with multidrug resistant HIV-1 infection by the US FDA and the European Medicines Agency in 2020 and 2021, respectively. In this review, we focus on the research progress of small molecule inhibitors targeting the interaction of gp120-CD4 from the perspective of medicinal chemistry, in order to provide reference for the subsequent research of gp120 inhibitors.

OBJECTIVETo study the human myxovirus resistant protein A (MxA), a specifically induced peptide by interferon I, and to use its level as a diagnostic criterion for viral infections.

METHODSAnti-MxA antisera from immunized mice were prepared with the expressed MxA protein of pET32a-MxA in E. coli BL-21(DE3). To confirm the antiserum activity and specificity, the expression product of BL21, wild type MxA pEGFP-C1-wMxA and site-directed mutant MxA pEGFP-C1-mMxA(N589S) stably transfected 3T3 cells and induced A549 cells were detected by Western blot with the antisera using non-MxA transfected or non-IFN-beta induced cells, intact A549, NIH 3T3 cells transfected with pEGFP-C1 and pET32a (+)-transformed BL-21 as controls.

RESULTSThe antisera had specific positive immunoreactivity to the NIH3T3 cells transformed with pEGFP-C1-wMxA and pEGFP-C1-mMxA, INF-beta induced A549 cells and BL21 proteins expressed with pET32a (+)-MxA. The hybridization signals from IFN-beta induced A549 cells depended on the IFN-beta inducing concentrations. Meanwhile, immunohistochemical assay showed that NIH 3T3 cells with pEGFP-C1-wMxA and pEGFP-C1-mMxA had > 98% of positive cells at 1:50 dilution of the serum and A549 cells induced by 20 ng/mL IFN-beta for 48 h showed 95% positive cells. pEGFP-C1-transfected NIH 3T3 cells were all negative.

CONCLUSIONAnti-sera are highly specific to diversified MxAs. The antibody is detectable by Western blot, immunocytochemistry and immunofluorescence assay.

Animals ; Antibody Specificity ; Cell Line, Tumor ; GTP-Binding Proteins ; genetics ; immunology ; metabolism ; Gene Expression Regulation ; Humans ; Mice ; Myxovirus Resistance Proteins ; NIH 3T3 Cells ; Species Specificity