Objective To construct the prostate-specific double gene expression vector pIRESPSMAe/p-TK-Cx43 and establish the foundation for experimental prostate cancer gene therapy research. Methods Cx43 gene was amplified and cloned into pMD19-T Simple vector. HSV-TK gene was then synthesized and cloned into multiple clone site (MCS) A of the eukaryotie vector plRES. The new plasmid was named plRES-TK: PSMAe/p was obtained and cloned into plRES-TK by replacing CMV promoter. The new plasmid was named plRES-PSMAe/p-TK; Fourth, Cx43 gene was cloned into the MCS B of pIRES-PSMAe/p-TK and the new plasmid was named pIRES-PSMAe/p-TK-Cx43.This plasmid was identified by double digestion with Sal Ⅰ/Not Ⅰ and sequenced; Finally, LNCaP cells were transfected by the plasmid plRES-PSMAe/p-TK-Cx43 and the mRNAs expression of HSV-TK gene and Cx43 gene was tested by RT-PCR. Results The plasmids synthesized in this experiment were double digested respectively and the specific bands of the inserted genes were confirmed by RTPCR. plRES-PSMAe/p-TK-Cx43 was in line with the expected design by DNA sequencing. The mRNAs of TK gene and Cx43 gene were expressed and successfully confirmed by RT-PCR after LNCaP cells transfected with pIRES-PSMAe/p-TK-Cx43. Conclusion Double gene expression vector pIRES-PSMAe/p-TK-Cx43 containing HSV-TK gene and Cx43 gene is constructed successfully.

BACKGROUND:Percutaneous vertebroplasty and percutaneous kyphoplasty have become the mainstream clinical methods for the treatment of vertebral compression fractures. However, both of them have several shortcomings such as bone cement leakage, spinal stenosis, nerve compression, pulmonary embolism and other issues. OBJECTIVE:To verify the possibility of bone filing mesh container prepared by polyethylene terephthalate for the treatment of vertebral compression fractures. METHODS:The biological properties of bone filing mesh container were examined according to GB/T16886. After sample aging test, the tensile properties of the aged samples and the fresh prepared samples were compared. The expansion and bone cement leakage were evaluated by injecting bone cement into the bone filing mesh container and measuring the pressure. The initial strength and stiffness of the fresh pig vertebrae with calcium phosphate cement injection or with bone filing mesh container filed with calcium phosphate cement were compared. The in vivo bone tissue growth was periodicaly observed after the lumbar vertebra of 4-month-old pigs was implanted with the bone filing mesh container that was then ful of bone cement. RESULTS AND CONCLUSION: The bone filing mesh container had good biocompatibility. Bone filing mesh containers after 2-year storage had the same tensile strength to the fresh bone filing mesh containers. At ambient conditions, after bone cement injection, bone filing mesh containers could be expanded at 5-10 atm and therefore could play the role of uplift; at 7-10 atm, bone cement could leak out from the bone filing mesh container and enter into the interspace between surrounding bone tissues, thus playing the role of adhesion and fixation. The vertebrae after bone cement injection with or without bone filing mesh containers had the same initial strength and stiffness and exhibited bigger initial strength and stiffness than untreated vertebrae. Thein vivo animal experiments proved that bone filing mesh container had no obvious effect on the vertebrae. These findings indicate that the bone filing mesh container can be used to restore the height and strength of the fractured vertebrae. Moreover, it may eliminate bone cement leakage and therefore increase the surgery safety.

OBJECTIVE: To compare the influence of traditional Chinese compound recipes (TCCRs) with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. METHODS: H(22) cancer-bearing mice were chosen to observe the effects of TCCRs with different efficacy on tumor growth inhibition and detect the proliferation function of T lymphocytes, the activity of natural killer (NK) cells, the changes of T lymphocytes and the content of interferon-gamma (IFN-gamma)and interleukin-4 (IL-4). RESULTS: Tumor weight of H(22) cancer-bearing mice in Yidu Gongdu Recipe (YDGDR, a compound traditional Chinese herbal medicine using poison as an antidote for poison)-treated group was obviously lighter than that in the other TCCR-treated groups and the tumor inhibition rate in YDGDR-treated group was 65.76% (P<0.01). The tumor inhibition rates in other TCCR-treated groups were ranged from 10.1% to 17.1% . Body weight of mice in YDGDR-treated group was obviously decreased and depilation was observed at the same time. Pelage of mice in Fuzheng Peiben Recipe (FZPBR, a compound traditional Chinese herbal medicine for supporting the healthy energy)-treated group grew well, and behavior of the mice was active. Stimulation index (SI) of T lymphocyte transformation in YDGDR-treated group was obviously increased (SI=4.34, P<0.01), which showed the proliferation function of T lymphocyte was very strong. The SI of T lymphocyte transformation in the other groups was less than three, which showed the proliferation function of T lymphocytes was not significant. Compared with normal saline (NS)-treated group, percentages of NK cells in Qinre Jiedu Recipe (QRJDR, a compound traditional Chinese herbal medicine for clearing away heat and toxic substances)-treated, Huxue Huayu Recipe (HXHYR, a compound traditional Chinese herbal medicine for activating blood circulation to dissipate blood stasis)-treated and YDGDR-treated groups were obviously increased and 5.05, 4.07 and 5.17 times more than the NS-treated group, respectively (P<0.01). The activity of NK cells wasn't increased in the FZPBR-treated and HXHYR-treated groups. The production of IFN-gamma induced by T cells in YDGDR-treated group was obviously raised (P<0.05), and the production of IL-4 induced by T cells in QRJDR-treated, HXHYR-treated, Huatan Sanjie Recipe (a compound traditional Chinese herbal medicine for eliminating phlegm and resolving masses)-treated and YDGDR-treated groups was also raised obviously (P<0.01). CONCLUSION: YDGDR has a good effect of inhibiting tumor growth and can reinforce cellular and humoral immune function in tumor-bearing mice. FZPBR can strengthen the body.

OBJECTIVE: To investigate the mechanisms of tumor inhibiting and immunoloregulation of Mylabris Mixture on H22 cancer-bearing mice. METHODS: H22 cancer-bearing mice were chosen to observe the effects of tumor inhibiting and detect the proliferation function of T lymphocytes, the toxicity function of NK cells, the changes of T lymphocytes and the contents of interferon-gamma and interleukin-4. RESULTS: Mylabris Mixture could obviously inhibit the growth of H22 cancer in mice, and the tumor inhibition rat was 65.76%. The stimulation index of T lymphocyte transformation and percentage of NK cells in Mylabris Mixture-treated group were obviously higher than those in the normal control group. The subpopulation proportion of T lymphocytes in Mylabris Mixture-treated group was changed more than the normal control group. The production of interferon-gamma and interleukin-4 by T lymphocytes obviously increased in Mylabris Mixture-treated group (P<0.05, P<0.001). CONCLUSION: Mylabris Mixture has the effect of inhibiting the growth of tumor constitution, and regulating immunological function on mice with tumor. Its mechanisms include the reinforcement of T lymphocyte immune function, NK cell killing function and humoral immune function.

Objectives: . Otitis media is a common infection worldwide. Owing to the limited number of ear specialists and rapid development of telemedicine, several trials have been conducted to develop novel diagnostic strategies to improve the diagnostic accuracy and screening of patients with otologic diseases based on abnormal otoscopic findings. Although these strategies have demonstrated high diagnostic accuracy for the tympanic membrane (TM), the insufficient explainability of these techniques limits their deployment in clinical practice. Methods: . We used a deep convolutional neural network (CNN) model based on the segmentation of a normal TM into five substructures (malleus, umbo, cone of light, pars flaccida, and annulus) to identify abnormalities in otoscopic ear images. The mask R-CNN algorithm learned the labeled images. Subsequently, we evaluated the diagnostic performance of combinations of the five substructures using a three-layer fully connected neural network to determine whether ear disease was present. Results: . We obtained the receiver operating characteristic (ROC) curve of the optimal conditions for the presence or absence of eardrum diseases according to each substructure separately or combinations of substructures. The highest area under the curve (0.911) was found for a combination of the malleus, cone of light, and umbo, compared with the corresponding areas under the curve of 0.737–0.873 for each substructure. Thus, an algorithm using these five important normal anatomical structures could prove to be explainable and effective in screening abnormal TMs. Conclusion . This automated algorithm can improve diagnostic accuracy by discriminating between normal and abnormal TMs and can facilitate appropriate and timely referral consultations to improve patients’ quality of life in the context of primary care.

High degree atrioventricular block (HDAVB) is a serious complication of transcatheter closure of a perimembranous ventricular septal defect (PMVSD). We report one patient who developed transient HDAVB seven days after transcathter closure of PMVSD and had recurrent HDAVB 42 months after the procedure.
Atrioventricular Block ; etiology ; Heart Septal Defects, Ventricular ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Recurrence ; Septal Occluder Device

High degree atrioventricular block (HDAVB) is a serious complication of transcatheter closure of a perimembranous ventricular septal defect (PMVSD). We report one patient who developed transient HDAVB seven days after transcathter closure of PMVSD and had recurrent HDAVB 42 months after the procedure.

OBJECTIVETo investigate the correlation between expression of A-kinase anchoring protein 95 (AKAP95) and protein expression of cyclin E1 and cyclin D1 in lung cancer tissue.

METHODSFifty-one cases of lung cancer were included in the study. The protein expression of AKAP95, cyclin E1, and cyclin D1 were measured by immunohistochemistry.

RESULTSThe protein expression of cyclin E1 in lung cancer tissues was significantly higher than that in para-cancerous tissues (positive rate: 75.56%vs 20%, P < 0.01); its expression showed no relationship with histopathological type, lymph node metastasis, and cellular differentiation (P > 0.05). The protein expression of cyclin D1 in lung cancer tissues was higher than that in para-cancerous tissues (positive rate: 69.39% vs 14.29%); its expression showed a significant relationship with histopathological type (P < 0.05). The expression of AKAP95 was correlated with the protein expression of cyclin E1 and cyclin D1 in lung cancer tissues (P < 0.01).

CONCLUSIONCyclin E1 and cyclin D1 are highly expressed in lung cancer tissue, suggesting that they play an important role in the development and progression of lung cancer. The protein expression of cyclin E1 has no relationship with cellular differentiation, lymph node metastasis, and histopathological type of lung cancer, and the protein expression of cyclin D1 has a significant relationship with histopathological type. The expression of AKAP95 is correlated with the protein expression of cyclin E1 and cyclin D1 in lung cancer tissue.

A Kinase Anchor Proteins ; metabolism ; Adult ; Aged ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Middle Aged ; Oncogene Proteins ; metabolism

OBJECTIVETo investigate the changes of endoplasmic reticulum stress-induced apoptosis in pulmonary tissue of rats with hypoxic pulmonary hypertension.

METHODSTwenty two male SD rats were randomly divided into control group and 4-week hypoxia-hypercapnia group (n=11). The mean pulmonary arterial pressure (mPAP) and the mean carotid arterial pressure (mCAP) were monitored, and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV + S) were measured. The rattish pathological model were assessed by mPAP, mCAP, RV/(LV+ S), vessel wall area/total area (WA/TA), vessel cavity area/total area (CA/TA) and media thickness of pulmonary arteriole (PAMT). The pulmonary apoptotic cells were detected by Hoechst staining. RT-PCR was used to study the genetic expression of caspasel2, glucose regulated protein 78 (GRP78) and GRP94 in pulmonary tissue. The expression of GRP94 and GRP78 proteins in pulmonary tissue were determined by using immunohistochemistry.

RESULTS(1) (The mPAP, RV/(LV + S), WA/TA and PAMT were respectively higher by 50.5%, 37.3%, 72.5% and 137% in hypoxic group than those in control group, while CA/TA was lower by 41.9% (all P < 0.01). There was not significant difference of mCAP between the two groups. (2) Hoechst staining showed that the pulmonary apoptotic cells in hypoxic group outnumbered markedly than those in control group, and the apoptotic cells were mainly in pulmonary tissue, while they were rare in pulmonary vascular smooth muscle cell. (3) Compared with control group, the expression of pulmonary caspasel2, GRP78 and GRP94 mRNA in hypoxic group were higher by 144%, 137% and 80.7% (all P < 0.05), respectively. (4) The expression of pulmonary GRP78 and GRP94 proteins were up-regulated in hypoxic group, and these proteins mainly localized in pulmonary vascular endothelial cell.

CONCLUSIONThe endoplasmic reticulum stress-induced apoptosis may be one of the mechanism of hypoxic pulmonary hypertension and pulmonary vascular wall remodeling.

Animals ; Apoptosis ; physiology ; Caspase 12 ; metabolism ; Endoplasmic Reticulum Stress ; physiology ; Heat-Shock Proteins ; metabolism ; Hypercapnia ; physiopathology ; Hypertension, Pulmonary ; etiology ; pathology ; physiopathology ; Hypoxia ; complications ; physiopathology ; Lung ; pathology ; Male ; Membrane Glycoproteins ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the expression of Rab11 in children with sepsis at different sta-ges and severe sepsis and its relationship with the occurrence and development of sepsis in children. Methods A prospective control study was performed. All cases were enrolled from Bao′an Maternal and Children Health Care Hospital, and they were divided into sepsis group(40 cases) who were diagnosed as sepsis, severe sepsis group(20 cases) with diagnosis of severe sepsis,and healthy control group(40 healthy chil-dren) . Venous blood samples were collected at admission,and the expression level of blood leukocyte Rab11 was determined by Western blot. In the sepsis group,the expression levels of Rab11 were evaluated at the initial,the extreme and the recovery stages of sepsis,and were compared with those in severe sepsis group and healthy control group, respectively. Spearman correlation analysis was used to evaluate the relationship between the expressions of Rab11 and the levels of some parameters in blood,including white blood cell,neu-trophils,lymphocytes,monocytes,eosinophilic granulocyte,C-reactive protein and procalcitonin in blood,at the extreme stage. Meanwhile,the levels of Rab11 at extreme stage of sepsis,caused by different diseases, such as severe pneumonia,bronchiectasis complicated pulmonary infection,biliary tract infection,urinary tract infection, necrotizing enterocolitis and severe enteric viruses infection, were compared with each other. Results At the initial and the extreme stages of sepsis, as well as in severe sepsis group,the levels of Rab11 were significantly lower than that in the healthy control group(0. 54 times,0. 23 times and 0. 07 times, P<0. 05,respectively). There were no significant differences in the expression levels of Rab11 at the recovery stages of sepsis compared with that in the healthy control group(P>0. 05). There was no relation-ship between the level of Rab11 and the number of white blood cell, neutrophils, lymphocytes, monocytes, eosinophilic granulocyte, while the level of Rab11 was negatively correlated with C-reactive protein ( r =-0. 58,P=0. 014) and procalcitonin(r= -0. 63,P=0. 003) at the extreme stage of sepsis. There was no significant difference in the expression level of Rab11,at the extreme stage of sepsis,among those patients with severe pneumonia,bronchiectasis and pulmonary infection,biliary tract infection,urinary tract infection, necrotizing enterocolitis and severe enterovirus infection(P>0. 05). Conclusion The level of Rab11 is differently expressed at different stages of sepsis,and could be used as a predictor of the severity of sepsis in children.