ABSTRACT
As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Subject(s)
Animals , Swine , Induced Pluripotent Stem Cells/metabolism , Receptors, Cell Surface/genetics , Antigens, CD/metabolism , Porcine respiratory and reproductive syndrome virus/geneticsABSTRACT
Objective To investigate the efficacy of acupuncture and rehabilitation therapy on lower limb motor function, and to explore a cortical mechanism using functional near infrared spectroscopy (fNIRS). Methods From December, 2020 to July, 2021, 24 stroke patients with lower limb motor dysfunction in our hospital were randomly divided into rehabilitation group (n = 12) and acupuncture-rehabilitation group (n = 12), and received routine rehabilitation training and acupuncture-rehabilitation intervention for four weeks, respectively. The control group included ten healthy subjects matched the patients. Before and after intervention, the lower limb motor function of the patients was assessed with Fugl-Meyer Assessment-Lower Extremities (FMA-LE), and all the subjects accepted fNIRS examination. The functional intensity and lateralization index (LI) of supplementary motor area (SMA), premotor cortex (PMC) and sensory motor cortex (SMC) were calculated based on oxygenated hemoglobin (HbO2). Results There was no significant difference in FMA-LE score between the rehabilitation group and the acupuncture-rehabilitation group before the intervention (P > 0.05). After four weeks of intervention, FMA-LE scores improved in both groups (t > 3.770, P 0.05). After intervention, the average functional connection increased in both groups (t > 2.178, P < 0.05), and the functional connection of the affected PMC of acupuncture-rehabilitation group increased (P < 0.05). The LI in SMC increased in the acupuncture-rehabilitation group (P < 0.05). There was a significant positive correlation between the change of functional connection of the affected PMC and the change of FMA-LE scores in the acupuncture-rehabilitation group (r = 0.579, P < 0.05). Conclusion Acupuncture with rehabilitation therapy can significantly improve the lower limb motor function and asymmetrical activation of SMC in stroke patients. The recovery of lower limb motor function may be related to the enhanced activation of affected PMC.
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Squamous cell carcinoma, in the clinical manifestation, is a form of cancer derived from lesions of keratinocytes of epidermis or accessories such as sebaceous ducts, hair follicles, sweat glands, etc. The disease is more common in older men, with prone locations at patients' scalp, face, neck and arms and other exposed parts. Head and neck squamous cell carcinoma [HNSCC] causes a serious impact on patients' daily life, making patients suffer from double blow in mental and physical aspects and reducing patients' life quality. To find effective treatment method for HNSCC, our hospital studies clinical effects of combination therapy of tegafur gimer, docetaxel and carboplatin for the disease. By way of grouping research, therapeutic effect of such treatment and adverse reactions were assessed and analyzed. The study clearly and fully confirms effectiveness of combination therapy of tegafur gimer, docetaxel and carboplatin for HNSCC
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Objective To explore the feasibility and safety of systemic chemotherapy concurrent whole brain radiotherapy (WBRT) for non-small cell lung cancer (NSCLC) with brain metastases.Methods Eighty cases of NSCLC patients with brain metastases were divided into observation group (40 cases) and the control group (40 cases) according to random number table.Patients were given systemic chemotherapy synchronous WBRT or sequential WBRT.Results The Ⅰ-Ⅳ degree leukocytopenia incidences of the two groups were 12.5％,25.0％,25.0％,0.0 and 30.0％,25.0％,12.5％,15.0％,and there was statistical significance (x2 =12.12,P ＜ 0.05).In the observation group,the total remission rate was 20％ (8/40),and it was 22.5％ (9/40) in the control group,with no significant difference (x2 =1.79,P ＞ 0.05).The median progression free survival of the observation group and of the control group were (3.5 ± 2.3) months and (3.6 ±1.1) months,respec-tively,with no significant difference (t =5.23,P ＞ 0.05).But the 1-years survival rate in the observation group was 37.5％ (15/40),significantly higher than that in the control group (17.5％,7/40),which had statistical significance (x2 =9.11,P ＜ 0.05).Conclusion The safety of systemic chemotherapy synchronous WBRT for NSCLC patients with brain metastases is higher,with good curative effect and strong application feasibility.
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Objective To investigate the expression of Annexin A1 proteins in colorectal cancer tissues and the relationship be-tween it and clinicopathologic features of colorectal cancer.Methods The surgical specimens from 48 cases of colorectal cancer,36 cases of adjacent tissues,15 cases of adenoma polyp were used to be examined expression of Annexin A1 immunohistochemical method.Results The positive expression rate of Annexin A1 in cancer tissue(64.6%)was significantly higher than those in adja-cent tissues(38.8%)and adenoma polyp tissues(40.0%)(P<0.05).The adjacent tissues and adenoma polyp tissues with positive Annexin A1 expression showed mild to severe atypical hyperplasia.The positive rates of Annexin A1 were significantly lower in the well-differentiated adenocarcinoma,tumor size of <5 cm,with no metastasis to lymphnodes and no invasion to surrounding tissues of colorectal cancer than those in the moderately or poorly differentiated adenocarcinoma,tumor size of ≥5 cm,with metastasis to lymphnodes and invasion in surrounding tissues(P<0.05).Conclusion The expression levels of Annexin A1 may be an important index of occurrence,progression and biological behaviors of colorectal carcinoma.
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Objective To evaluate the correlation of chemosensitivity of cancer cells and peripheral blood lymphocytes to 8 chemotherapy agents in vitro,which was helpful to guide clinical chemotherapy for patients with lung cancer. Methods The chemosensitivity of the peripheral blood lymphocytes and tumor cells to 8 clinical routine che-motherapeutic agents was tested in 40 cases of lung cancer by using MTT method. SPSS 10. 0 was used to analyze the above indicators. Results There was no significant difference in the sensitivity of 8 clinical routine chemotherapeutic agents between peripheral blood lymphocytes and tumor cells of patients with lung cancer(P > 0. 05) ,which was posi-tive correlated to chemotherapeutic drugs between peripheral blood lymphocytes and tumor cells. Conclusion The chemosensitive test of the peripheral blood lymphocytes with MTT method in vitro was valuable for reference of selec-ting anticancer drugs in clinic for patients with lung cancer.
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Objective Research on variable substitution to non-linear regression forecast model precision's influence, and seek the modelling method that can improve the forecast precision. Methods Based on the data mining,the transform in space and the weighted processing combined method, make full use of information that the primary data provide. Results Given modelling method of combination forecast model based on the data mining. Conclusion Based on data mining's combination forecast model's modelling method can reduce the serious influence that the variable substitution brings and has fully used useful information in the primary data. It obviously improved the accuracy of the prediction model.
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Objective:To study the disturbance creatd by the variable transformation within regression parameters of nonlinear biology models and find a new method removing or reducing the disturbance.Methods:To use the space transformation,Taylor series and weight method to remove or reduce the serious disturbance created by the variable transformation within regression parameters of nonlinear biology models.Results:Giving a new method calculating regression parameters of nonlinear biology models.Conclusion:The new method can remove or reduce the serious disturbance created by the variable transformation and remain the advantage of variable transformation.It can not only improve regression precision of the nonlinear biology models but also can find problems hidden in the original data.
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Exogenous gene suture was used to achieve peripheral nerve anastomoses to probe into the feasibility that the sites of anastomoses of nerves directly transfer gene and thus enable gene to be expressed at the sites of anastomoses under the condition that perfect nerve anastomoses are ensured. PCMVβ plasmid containing cytomegalovirus promoter (CMV promoter) and Escherichia coli (E.coli) β-Galactosidase (β-Gal) structural gene (lacZ gene) was conducted. A soaked medical 8-0nylon suture was used to perform epineurial repair of rabbit sciatic nerve. In the control group a suture soaked in sucrose PBS was used, while in the experimental group a suture soaked in PCMVβ plasmid solution was applied. The sites of anastomoses of nerves by stages were taken out, and β-Gal histochemical staining was performed and β-Gal enzyme activity was assayed with 5-bromo-4-chloro-3-indolyl-β-D-galactoside. Results showed that the sites of anastomoses of nerves were taken out 2 days, 7 days, 14 days and 30 days respectively after the operation. The β-Gal histochemical stains at the sites of anastomoses showed no indigo positive cells at different stages in the control group, whereas displayed indigo positive cells in the experimental group. In the control group, no β-Gal enzyme activity was detected at different stages after operation, but in the experimental group, β-Gal enzyme activity could be detected from the 3rd day to the 30th day after operation. It was concluded that by using exogenous gene suture, exogenous gene could be transferred to the sites of peripheral nerve and expressed the exogenous gene expression products with bioactivity, which provided the feasibility of using gene therapy to accelerate the recovery of nerve function.
ABSTRACT
Exogenous gene suture was used to achieve peripheral nerve anastomoses to probe into the feasibility that the sites of anastomoses of nerves directly transfer gene and thus enable gene to be expressed at the sites of anastomoses under the condition that perfect nerve anastomoses are ensured. PCMVβ plasmid containing cytomegalovirus promoter (CMV promoter) and Escherichia coli (E.coli) β-Galactosidase (β-Gal) structural gene (lacZ gene) was conducted. A soaked medical 8-0nylon suture was used to perform epineurial repair of rabbit sciatic nerve. In the control group a suture soaked in sucrose PBS was used, while in the experimental group a suture soaked in PCMVβ plasmid solution was applied. The sites of anastomoses of nerves by stages were taken out, and β-Gal histochemical staining was performed and β-Gal enzyme activity was assayed with 5-bromo-4-chloro-3-indolyl-β-D-galactoside. Results showed that the sites of anastomoses of nerves were taken out 2 days, 7 days, 14 days and 30 days respectively after the operation. The β-Gal histochemical stains at the sites of anastomoses showed no indigo positive cells at different stages in the control group, whereas displayed indigo positive cells in the experimental group. In the control group, no β-Gal enzyme activity was detected at different stages after operation, but in the experimental group, β-Gal enzyme activity could be detected from the 3rd day to the 30th day after operation. It was concluded that by using exogenous gene suture, exogenous gene could be transferred to the sites of peripheral nerve and expressed the exogenous gene expression products with bioactivity, which provided the feasibility of using gene therapy to accelerate the recovery of nerve function.