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OBJECTIVE To investigate the effect and mechanism of Panax notoginseng saponins （PNS） on wound healing after anal fistula surgery in rats by regulating the hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor （VEGF）/ vascular endothelial growth factor receptor-2 （VEGFR2） signaling pathway. METHODS SD rats were selected to establish a postoperative rat model of anal fistula by infecting wound with Escherichia coli. The model rats were randomly grouped into model group， PNS low-dose and high-dose groups （15， 30 mg/cm2）， high-dose of PNS+2-methoxyestradiol （2ME2） group （PNS 30 mg/cm2+HIF-1α inhibitor 2ME2 4 mg/kg）， with 10 rats in each group. Another 10 normal rats were selected for back hair removal treatment as the control group. Each drug group was injected with the corresponding drug solution intramuscularly or （and） intraperitoneally， once a day， for 3 weeks. After the last administration， the wound healing rate （excluding the control group）， microvascular density （MVD）， the expression of collagen Ⅰ and fibronectin （FN） in the wound tissue were detected in each group； the levels of angiogenic factors [VEGF， E-mail：842710813@qq.com angiopoietin-Ⅰ （Ang-Ⅰ）， Ang-Ⅱ] in serum， the levels of inflammatory factors [interleukin-6 （IL-6） and IL-2] in serum binggui7183@163.com and wound tissue as well as the expressions of the related proteins of HIF-1α/VEGF/VEGFR2 signaling pathway in the wound tissue of rats were also detected in each group. RESULTS The MVD， the expression of collagen Ⅰ and FN in the wound tissue， and the levels of IL-6 and IL-2 in serum and wound tissue of rats increased significantly in the model group， compared to the control group （P＜0.05）， while the serum levels of VEGF， Ang- Ⅰ and Ang-Ⅱ decreased significantly （P＜0.05）. The wound healing rate， the MVD in wound tissue， the serum levels of VEGF， Ang-Ⅰ and Ang-Ⅱ， the expressions of collagen Ⅰ and FN in the wound tissue， and protein expressions of HIF-1α， VEGF and VEGFR2 in the PNS low-dose and high-dose groups increased significantly， compared to the model group （P＜0.05）， while the levels of IL-6 and IL-2 in serum and wound tissue decreased significantly （P＜0.05）； the high-dose PNS had a stronger effect （P＜ 0.05）. 2ME2 could weaken the effect of PNS on above indicators of rats after anal fistula surgery （P＜0.05）. CONCLUSIONS PNS can promote the production of angiogenic factors and inhibit the production of pro-inflammatory factors， thereby promoting wound healing in rats after anal fistula surgery. The above effects are related to the activation of HIF-1α/VEGF/VEGFR2 signaling pathway.
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BACKGROUND:Traumatic patellar dislocation with medial patellofemoral ligament tearing at femoral attachment or body is usually performed by medial patellofemoral ligament reconstruction surgery.To promote tendon bone healing after medial patellofemoral ligament reconstruction,the researchers used a variety of biological treatment technologies including growth factors,stem cells and platelet-rich plasma. OBJECTIVE:To investigate the clinical effect of medial patellofemoral ligament reconstruction by leukocyte-and platelet-rich fibrin with autologous hamstring tendon for traumatic patellar dislocation. METHODS:Thirty-seven patients with traumatic patellar dislocation in First Hospital of Qinhuangdao from February 2019 to February 2021 were randomly divided into a trial group(n=18)and a control group(n=19).The trial group received medial patellofemoral ligament reconstruction by leukocyte-and platelet-rich fibrin with an autologous hamstring tendon.The control group received medial patellofemoral ligament reconstruction by a simple autologous hamstring tendon.Patients in the two groups were followed up for 12 months.Knee pain and functional status were evaluated by visual analog scale score,Lysholm score,Kujala patellofemoral joint score and knee range of motion.The patellar tilt angle,patellar congruence angle and patellar lateral shift rate of the patellofemoral joint were measured by MRI and CT films to evaluate the stability and improvement of the patellofemoral joint. RESULTS AND CONCLUSION:(1)The visual analog scale scores of the two groups at 6 and 12 months after operation were lower than those before operation(P<0.05).The Lysholm score and Kujala patellofemoral joint score at 6 and 12 months after operation were higher than those before operation(P<0.05).The Lysholm score and Kujala patellofemoral joint score in the trial group were higher than those in the control group 6 months after operation(P<0.05).There was no significant difference between the two groups in the visual analog scale score,Lysholm score and Kujala patellofemoral joint score 12 months after operation(P>0.05).(2)The patellar tilt angle,patellar congruence angle,patellar lateral shift rate and range of motion of the patellofemoral joint were significantly improved in both groups 12 months after operation(P<0.05).The patellar tilt angle was smaller in the trial group than that in the control group 12 months after operation(P<0.05).Patellar congruence angle,patellar lateral shift rate,range of motion and MRI score were not statistically significant between the two groups 12 months after operation(P>0.05).(3)These results confirm that medial patellofemoral ligament reconstruction by leukocyte-and platelet-rich fibrin with autologous hamstring tendon can treat traumatic dislocation effectively,improve the function of the knee joint,and restore the movement track of the patella.
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Herein, the effect of Fushun oil shale minerals on its kerogen has been investigated. The samples were obtained under non-isothermal conditions at different final temperatures. Scanning electron microscope (SEM) analysis revealed that the silicates were layered and the carbonates were tightly bound to each other. The combination of silicates and carbonates led to close combination of minerals and organic matter and the organic matter was contained in the minerals. The Brunner-Emmet-Teller (BET) experiments conclude that during the non-isothermal pyrolysis process, the specific surface area increased, and then, decreased, which proves the adsorption effect of silicates on oil shale pyrolysis products and the adsorption effect of carbonates was weak. The activation energy of four samples was calculated via Flynn-Wall-Ozawa (FWO) and Friedman kinetic analysis under different heating rates in a non-isothermal process, wherein the average activation energy of the sample containing silicate was 177.60 kJ mol-1 at minimum while that of carbonate was 250.45 kJ mol-1 at maximum, which proves that the catalytic promotion effect of silicate was greater than the inhibition effect of carbonate. The pyrolysis products obtained by Flash pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) under isothermal pyrolysis conditions were primarily composed of aliphatic hydrocarbon structures, which had different degrees of impact on the production of heteroatoms. This work provides a reliable theoretical basis for future studies on the influence of minerals on pyrolysis of organic matter in oil shale.
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Objective:To compare the short-term clinical effects of absorbable cross nail, sheath compression nail and suspension fixation on the femoral side and sheath compression nail on the tibial side in anterior cruciate ligament reconstruction (ACLR) with autologous hamstring tendon.Methods:The clinical data of 373 patients with anterior cruciate ligament injuries treated with surgery in the First Hospital of Qinhuangdao Affiliated to Chengde Medical College from December 2008 to December 2018 were collected for a retrospective case-control study. There were 125 cases of absorbable cross nail (absorbable cross nail group), 112 cases of sheath compression nail (sheath compression nail group) and 136 cases of suspension fixation (suspension fixation group). The mean follow-up time was (13.96±1.42) months. The international knee documentation committee (IKDC) score and Lysholm score were used to score the patients, and the differences were statistically analyzed. The measurement data of normal distribution was represented by xˉ± s, and repeated measurement analysis of variance was used to compare the three groups in different time periods, and the operation time of the three groups of samples was compared by one-way analysis of variance. The counting data were compared by χ 2 test. Results:(1) There were no complications such as infection, adhesion, stiffness and neurovascular injury in the three groups at the last follow-up. (2) Comparison of preoperative and postoperative knee scores: Lysholm score: preoperative absorbable cross nail group (65.98±1.37), sheath compression nail group(66.13±2.13), suspension fixation group (65.76±1.55). Three months after operation, absorbable cross nail group (80.14±2.02), sheath compression nail group (79.70±1.98) and suspension fixation group (79.84±1.86). Twelve months after operation, there were (94.56±2.35) points in the absorbable cross nail group, (94.96±1.34) points in the sheath compression nail group and (94.40±3.15) points in the suspension fixation group.There was significant difference in the scores before and after operation in the same group ( Fintra-group=17 584.14, P<0.001), and there was no significant difference among the three groups ( Finter-group=2.65, P=0.072), There was no statistical significance in the trend of scores before and after operation in the three groups ( Finter-action=1.28, P=0.277). IKDC scores: Preoperative (62.02±1.43) in absorbable cross nail group, preoperative (61.95±0.82) in sheath compression nail group, preoperative (62.25±2.05) in the suspension fixation group. Three months after operation, absorbable cross nail group (77.99±2.18), sheath compression nail group (78.13±2.02) and suspension fixation group (77.97±2.24). Twelve months after operation, the absorbable cross nail group (92.68±3.21), the sheath compression nail group (93.25±2.04) and the suspension fixation group (92.96±3.11). There was significant difference in the scores before and after operation in the same group ( Fintra-group=18 338.15, P<0.001), and there was no significant difference among the three groups ( Finter-group=0.91, P=0.402), and there was no significant trend in the scores before and after operation in the three groups ( Finteraction=0.98, P=0.419). (3) Comparison of operation time: absorbable cross nail group (99.04±18.01) min, sheath compression nail group (112.88±19.79) min and suspension fixation group (83.81±16.69) min. there was significant difference among the three groups ( F=79.74, P<0.001). Further comparison between the three groups, the time of suspension fixation group was the shortest than the other two groups (all P<0.001). Conclusion:There is no significant difference in the short-term efficacy of ACLR with autologous hamstring tendon among the three groups, and there were no complications such as infection, adhesion, stiffness and neurovascular injury in the last follow-up. In terms of operation time, the suspension fixation group has the shortest time and is simple and easy to operate.
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Severe acute respiratory syndrome CoV-2 (SARS-CoV-2) caused the Corona Virus Disease 2019 (COVID-19) cases in China has become a public health emergency of international concern (PHEIC). Based on angiotensin converting enzyme 2 (ACE2) as cell entry receptor of SARS-CoV, we used the hACE2 transgenic mice infected with SARS-CoV-2 to study the pathogenicity of the virus. Weight loss and virus replication in lung were observed in hACE2 mice infected with SARS-CoV-2. The typical histopathology was interstitial pneumonia with infiltration of significant lymphocytes and monocytes in alveolar interstitium, and accumulation of macrophages in alveolar cavities. Viral antigens were observed in the bronchial epithelial cells, alveolar macrophages and alveolar epithelia. The phenomenon was not found in wild type mice with SARS-CoV-2 infection. The pathogenicity of SARS-CoV-2 in hACE2 mice was clarified and the Kochs postulates were fulfilled as well, and the mouse model may facilitate the development of therapeutics and vaccines against SARS-CoV-2.
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The damage behaviors of polyimide fiber after 150 keV proton irradiation and the synergistic effect of proton irradiation and strain were investigated. Changes in the mechanical properties, free radicals, element content, and element chemical state of the polyimide fiber before and after 150 keV proton irradiation were investigated. The results showed that the tensile strength and elongation at break of the material decreased significantly after proton irradiation. The synergistic effect of proton irradiation and strain weakened the reduction of mechanical properties caused by single proton irradiation. After proton irradiation and the combination of proton irradiation and strain, pyrolytic carbon free radicals were generated. According to XPS analysis, the proton-irradiated polyimide fiber underwent complex denitrification and deoxygenation reactions, and carbon enrichment appeared on the surface of the material.
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explore the therapeutic effect of modified pedicled nasal septum mucosa flap on cerebrospinal fluid leakage in transsphenoidal pituitary surgery. Methods The clinical data of 28 patients treated with modified pedicled nasal septal mucosa flap during endoscopic transsphenoidal resection of pituitary adenoma in Dalian Central Hospital from August 2017 to December 2018 were analyzed retrospectively. Seven cases of high-flow cerebrospinal fluid leakage occurred during the operation. Skull base reconstruction was performed with modified pedicled nasal septum mucosal flap. Modified pedicled nasal septum mucosal flap was repositioned in 21 cases. Results Seven cases of modified pedicled nasal septum mucosal flap for skull base reconstruction had no postoperative cerebrospinal fluid leakage and 1 case had hypoolusia. Postoperative cerebrospinal fluid leakage occurred in One of the 21 patients with mucosal flap replacement, and stopped after 8 d of continuous lumbar drainage. One case had hypoolusia. Conclusions Modified pedicled nasal septum mucosal flap is a safe and reliable technique. It can not only achieve the effect of pedicled nasal septum mucosal flap in skull base reconstruction, but also avoid nasal complications caused by excessive application of mucosal flap.
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Objective@#To explore the therapeutic effect of modified pedicled nasal septum mucosa flap on cerebrospinal fluid leakage in transsphenoidal pituitary surgery.@*Methods@#The clinical data of 28 patients treated with modified pedicled nasal septal mucosa flap during endoscopic transsphenoidal resection of pituitary adenoma in Dalian Central Hospital from August 2017 to December 2018 were analyzed retrospectively. Seven cases of high-flow cerebrospinal fluid leakage occurred during the operation. Skull base reconstruction was performed with modified pedicled nasal septum mucosal flap. Modified pedicled nasal septum mucosal flap was repositioned in 21 cases.@*Results@#Seven cases of modified pedicled nasal septum mucosal flap for skull base reconstruction had no postoperative cerebrospinal fluid leakage and 1 case had hypoolusia. Postoperative cerebrospinal fluid leakage occurred in One of the 21 patients with mucosal flap replacement, and stopped after 8 d of continuous lumbar drainage. One case had hypoolusia.@*Conclusions@#Modified pedicled nasal septum mucosal flap is a safe and reliable technique. It can not only achieve the effect of pedicled nasal septum mucosal flap in skull base reconstruction, but also avoid nasal complications caused by excessive application of mucosal flap.
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Objective To explore the therapeutic effect of pedicled nasal septum mucosal flap on high-flow cerebrospinal fluid leakage in transsphenoidal approach. Methods The clinical data of 31 patients with high-flow cerebrospinal fluid leakage during neuroendoscope transsphenoidal approach from January 2012 to April 2018 were analyzed retrospectively. Among them, skull base of 18 patients was reconstructed with pedicled nasal septum mucosal flap technique (observation group), and skull base of 13 patients was reconstructed with the'sandwich'method (control group). The postoperative cerebrospinal fluid leakage and complications were compared between 2 groups. Results Postoperative cerebrospinal fluid leakage occurred in 6 cases in control group, and 1 case in observation group, and there was statistical difference between 2 groups (P<0.05). Postoperative olfactory loss occurred in 2 cases in control group, and 3 cases in observation group, and there was no statistical difference between 2 groups (P>0.05). Conclusions Multilayer skull base reconstruction with pedicled nasal septum mucosal flap can significantly reduce the incidence of cerebrospinal fluid leakage after transsphenoidal tumor resection, and is a safe and reliable method to treat the high flow cerebrospinal fluid leakage in operation.
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Objective To evaluate the monitoring effect of microvascular Doppler blood flow detector in intracranial aneurysm surgery. Methods One hundred and twenty patients with intracranial aneurysms from July 2015 to July 2017 who received craniotomy and used microvascular Doppler ultrasound 1 mm microprobe to detect the hemodynamics of the parent artery and related perforating artery change were retrospectively analyzed . According to the blood flow signal, aneurysm clip was not adjusted in 55 cases; when the tumor artery or perforator artery blood flow signal decreased or disappeared after treatment, aneurysm clip clamping position was changed in 65 cases, so that the blood flow of tumor artery or perforator artery returned to normal. The blood flow velocity and pulsation index (PI) of the artery was measured before and after adjustment of the aneurysm clip. The patients underwent CT angiography (CTA) or DSA examination 1 week after surgery. Angiography was performed to evaluate the postoperative results in one month, three months, and six months' follow-up. The data obtained were statistically analyzed. Results Vascular obstruction or neck residual rate of 120 cases was lower than 5％vascular obstruction or residual neck residual rate in large cases. Conclusions The microvascular Doppler blood flow detector is safe, reliable and easy to operate. This technique is useful for finding the clips in the craniotomy of the intracranial aneurysm or the incomplete clamping of the tumor-bearing artery, so as to adjust the position of the aneurysm clip in time and improve the curative effect.
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Objective To investigate the results of arthroscopic anterior cruciate ligament anatomic reconstruction using three-tunnel double-bundle methods with Femoral-Intrafix system.Methods From October 2010 to October 2013,data of 30 cases of arthroscopic anterior cruciate ligament (ACL) reconstruction who were randomly used with Femoral-Intrafix system were retrospectively analyzed.Hamstring tendons autograft were used for all patients.The Femoral-Intrafix system fixation was used for the femoral tunnel and the Milagro interference screw was used for tibial tunnels.KT-1000 arthrometer measurements,Lachman and Pivot-shift tests,and knee function with the Intemational Knee Documentation Committee (IKDC),Lysholm scores were used for clinical evaluation.Results All the patients were followed up for an average time of 24.71 ± 1.60 months (range,18-36 months).No patients had ligament re-rupture,deep vein thrombosis,exfliation of interference screw,joint pain and limited mobility during follow-up period.At the latest follow-up,the Lachman sign was Ⅰ degrees positive in 1 patient,and all patients had normal range of motion.The Lysholm scores increased from preoperative 67.51 ±4.92 to 96.41 ±6.31 at the latest follow up.The IKDC scores increased from preoperative 62.31 ±5.45 to 95.61±6.32 at the latest follow up.There was 93.3％ excellent and good rate,including excellent in 23 cases,good in 5 cases,and fair in 2 cases according to the IKDC.All knee function scores were improved.The results of KT-1000 were improved significantly (7.62± 1.25 mm vs.1.53± 1.02 mm at 30° of flexion,5.43± 1.21 mm vs 1.07± 1.02 mm at 90° of flexion) at the tensile fore of 134 N.Conclusion Arthroscopic ACL reconstruction using three-tunnel double-bundle methods with the Femoral-Intrafix system fixation is a reliable technique,which is easy to operate and has satisfactory short clinical outcome for knee function recovery.
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AIM:To investigate the effect of inhibiting Mcl-1 gene expression on apoptosis of mouse peritoneal macrophages infected with different virulence of Mycobacterium tuberculosis using a technique of RNA interference .METH-ODS:The BALB/c mice were infected with prepared bacterium of the virulence strains of Xinjiang , H37Rv, H37Ra and BCG.Mcl-1-shRNA was applied to the mouse model of infection , and the control groups were set up .On 1 d, 3 d, 5 d and 7 d, the mouse peritoneal macrophages were collected .The expression of Mcl-1 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The apoptotic rate of peritoneal macrophages was analyzed by flow cytometry . RESULTS:The expression of Mcl-1 at mRNA and protein levels was up-regulated in the peritoneal macrophages from the mice infected with different virulence of Mycobacterium tuberculosis, and the cells from the mice infected with virulence strains of Xinjiang and H37Rv expressed higher level of Mcl-1 than the uninfected control cells (P<0.05).The expres-sion of Mcl-1 at mRNA and protein levels was reduced by RNA interference as compared with control group ( P<0.05 ) . Inhibition of Mcl-1 expression induced apoptosis of peritoneal macrophages in the mice .CONCLUSION: The Mcl-1 ex-pression at mRNA and protein levels in mouse peritoneal macrophages infected with different virulence of Mycobacterium tu-berculosis was effectively suppressed by Mcl-1-shRNA, which can induce macrophage apoptosis .
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[ ABSTRACT] AIM: To explore the effects of Mcl-1 signal pathway blockers on Mcl-1 expression, macrophage apoptosis and Mycobacterium tuberculosis in the model of mice infected with Mycobacterium tuberculosis H37Rv.METH-ODS:A mouse infection model was established by intraperitoneal injection of H37Rv suspension.The signaling pathway blockers AG490, PD98059 and LY294002 for JAK/STAT, MAPK and PI3K, respectively, were intraperitoneally injected into the mice infected with H37Rv.Cell acid-fast staining was used to observe whether the mouse peritoneal macrophages infected with H37Rv were successfully established.Immunocytochemical method was employed to detect Mcl-1 expression in the mouse peritoneal macrophages infected with H37Rv.The apoptotic rate in each group was measured by flow cytomer-ty.The scavenging capacity of apoptotic macrophages against H37Rv was determined by Mycobacterium tuberculosis colony counting.RESULTS:The result of cell acid-fast staining revealed the existence of dispersive arrangement of red short anti-acid Mycobacterium tuberculosis within infected macrophages.The result of cell immunocytochemistry showed strongly posi-tive expression of Mcl-1 protein in H37Rv infection group, AG490 treatment group and LY294002 treatment group, weakly positive expression of Mcl-1 protein in PD98059 treatment group, and negative expression of Mcl-1 protein in control group. The result of flow cytometry found that the macrophage apoptotic rate in H37Rv infection group was higher than that in con-trol group, while that in PD98059 treatment group was high than that in other groups with statistically significant differences (P<0.05).The result of Mycobacterium tuberculosis colony counting showed that PD98059 treatment had the most signifi-cant inhibitory effect on H37Rv strain.CONCLUSION: Mcl-1 signaling pathway blockers increase the apoptotic rate of macrophages infected with Mycobacterium tuberculosis H37Rv and inhibit the growth of Mycobacterium tuberculosis by inhibi-ting the signaling pathways of JAK/STAT, MAPK and PI3K, among which the MAPK has the most obvious interfering effect on Mcl-1, and leads to the highest apoptotic rate of infected macrophages and the strongest bacteriostasis.
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Objective:To transfect Mcl-1shRNA into Raw264.7 cells,and screen out specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene to figure out the effect of shRNA on Mcl-1 expression in murine macrophage cell line Raw 264.7.Methods: Specific shRNA was transfected into murine macrophage cell line Raw 264.7 via lipofectamine.Semi-quantitative RT-PCR and Western blot were respectively employed to test the changes in Mcl-1 mRNA level and Mcl-1 protein expressions 24 h and 48 h after the transfection ,and the silencing effects of the three pairs of specific shRNA fragments corresponding to different sites were analyzed.Results: Specific shRNA fragments at 24 h and 48 h could effectively reduce Mcl-1 mRNA and protein level ,with higher silencing effects than those of the normal group ,the lipofectamine group and the negative control group.There were statistically significant differences among them ( P<0.05 ).Among the three pairs of specific shRNA fragments corre-sponding to different sites ,Mcl-1 shRNA3 showed the most significant inhibiting effect on Mcl-1 mRNA and proteins.Conclusion:RNA interference can downregulate the level of Mcl-1 mRNA in murine macrophage cell line Raw 264.7 and greatly downregulate the expression of Mcl-1protein.Specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene have been screened out successfully.
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Aim To study the combined effect Euphor-bia kansui of and Glycyrrhiza uralensis on metabolism of Kansuinine A and Kansuinine B. Methods Control, GU and GU plus EK groups were treated for 10 days, respectively. Then the liver microsomes were pre-pared. KA and KB were incubated with microsomes of each group, and concentrations of KA and KB were measured to reflect the metablism of KA and KB. E-rythromycin, diphenhydramine hydrochloride,benzbro-marone, cimetidine, fluconazole the inhibitors of CYP3 A4 , CYP2 D6 , CYP2 C9 , CYP1 A2 , CYP2 C19 respectively, were incubated with KA and KB. Con-centrations of KA and KB were measured to reveal the cytochrome P450 isoforms which involved in the metab-olism of them. KA and KB were incubated with glycyr-rhizic acid and enoxolone. Then concentrations of KA and KB were measured to reveal the effects of glycyr-rhizic acid and enoxolone to KA and KB. Results Concentrations of KA and KB in GU plus EK group were significantly higher than those in control and EK groups, which showed that the metablisom of KA and KB was inhibited in GU plus EK group. In addition, concentrations of KA and KB were higher in microsomes incubated with erythromycin, diphenhydramine hydrochloride, benzbromarone, cimetidine and fluconazole than in control group, which revealed that the metablism of KA and KB was slowed down when CYP3A4, CYP2D6, CYP2C9, CYP1A2 and CYP2C19 were inhibited. They may participate in the metablisom of KA and KB. After incubation with glycyrrhizic acid and enoxolone, the metablism of KA and KB was slowed down. Conclusions KA and KB may be the substrates of CYP2C19. GU plus EK can inhibit the activity of CYP2C19, which resulted in KA and KB metablism slowing down and accumulation. In addition, glycyrrhizic acid and enoxolone can inhibit the metablism of KA and KB. It may be one of the reasons of increased toxicity of GU plus EK.
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Objective To study the expressions of myeloid cell leukemia-1 (Mcl-1 ) gene in mouse macrophages Raw264.7 and human macrophages THP-1,to screen out the cell lines with high levels of expression as the experimental cells,and based on the screening results to construct the short hairpin RNA(shRNA)eukaryotic expression plasmid targeting mice Mcl-1 gene for transfection and further screen out the shRNA expression plasmid with the most obvious effect in silencing Mcl-1 gene.Methods Semi-quantitative PCR method was used to detect the expression of Mcl-1 mRNA in the two kinds of macrophages.Western blotting was adopted to detect the expressions of Mcl-1 proteins in the two kinds of macrophages.Three different gene loci for Mcl-1 shRNA fragments were designed with small molecules interfering RNA (siRNA)software.Eukaryotic expression plasmid Mcl-1 shRNA 1-3 carrying the shRNA fragments was constructed by a company.And the eukaryotic expression plasmid vector was transfected into scavenger cells Raw264.7 mice via through the liposome.The transfection results 24 h and 48 h after transfection were observed under the inverted fluorescence microscope;Mcl-1 mRNA and protein expression were detected by real time quantitative PCR and Western blot,respectively.Results The relative expression levels of Mcl-1 mRNA and protein in mouse macrophages Raw264.7 were significantly higher than those of human macrophages (P <0.05).The shRNA expression vector constructed within 24 h and 48 h could decrease Mcl-1 mRNA and protein levels in Raw264.7 cells,especially with the most obvious silencing effect at 48 h.The 48-h transfection group differed significantly from normal group,liposome group and negative control group (P <0.05).Compared with Mcl-1shRNA1and Mcl-1shRNA2,Mcl-1shRNA3 had the strongest effect in silencing Mcl-1mRNA and protein.Conclusion We have successfully screened out experimental Raw264.7 cells and Mcl-1 shRNA eukaryotic expression plasmid which has an obvious silencing effect targeting on Mcl-1 in mice macrophages Raw264.7.
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Objective To study the beneficial effect and mechanism of Alcea rosea roots in nephrolithiasis model in-duced by 1%ethylene glycol in rats. Methods We randomly divided 60 male Wistar rats into six groups,including control group, model group, Alcea rosea roots lower-dose preventive group, Alcea rosea roots high-dose preventive group, Alcea ro-sea roots lower-dose curative group, Alcea rosea roots high-dose curative group. Control group was free to access food and water;model group was given 1%ethylene glycol drinking water and was fed with normal diet, preventive group was given 1%ethylene glycol drink and Alcea rosea roots in low (250 mg/kg) or high dose (500 mg/kg) each day, curative group re-ceived 1%ethylene glycol drink each day and Alcea rosea roots in low or high dose from day 15 to day 28. At the end of the experiment, various renal functional and injury markers such as urine volume, calcium, phosphate, magnesium, urea, creati-nine, and oxalate were evaluated using urine, serum, and kidney homogenates. The kidneys were removed and prepared for histological evaluation of calcium oxalate deposits. Results In model groups, urine output, urea, creatinine, 24 h urine Ca2+, and oxalate and MDA were increased compared to those in control group(P＜0.05). GSH and SOD were increased in preventative and curative groups compared to those in the model group(P ＜ 0.05). The urea, creatinine, 24 h urine Ca2+, urine oxalate, MDA were reduced in preventive and curative groups compared to those in the model group(P＜0.05). The number and size of calcium oxalate crystal deposits were also less and smaller, and the kidney damage was less severe in pre-ventive and curative groups compared to in the model group. Conclusion The extract of Alcea rosea roots can prevent and treat calcium oxalate urinary stone formation in rats and protect renal function.
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The aim of this study is to investigate the immune effects of BCG primary immunization and IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immunization on mice .We randomly divided the mice into 7 groups ,namely PBS negative controls ,BCG controls ,pcAg85A controls ,BCG primary immunization combined with Ag85A booster immunization controls ,BCG primary immunization combined with Ag85A and IL-12 booster immunization controls , BCG primary immunization combined with IL-12 booster immunization controls ,and BCG primary immunization combined with pcDNA3 .1 booster immunization controls .Implementing the immune in procedure of BCG primary immunization and cytokine IL-12 booster immunization combined with mycobacterium tuberculosis Ag85A DNA ,we observed the immune effect on mice by detecting the mice serum total IgG , specific lymphocyte proliferation and cytokine levels in 4 ,6 ,8 weeks after the last immunization .Comparing the mice immunized in the strategy of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine strengthening immunization to the mice in other groups by other immune ways ,we found that ,in BCG/Ag85A+ IL-12 groups ,IgG in-creased significantly (P＜ 0 .05) ,specific lymphocyte prolifera-ted significantly ,and after strengthening immunization IFN-γlevels ,IL-2 levels and IL-4 levels in the three periods were 128 .2 ±20.4,190.2±16.51,244.2±39.14 ;146.2±17.29,271.6±16.36and16.36±28.12 ;68.6±6.62,96.6±5.5and5.5± 10 .71 ,respectively ,which were higher than those in other groups (P＜0 .05) .It’s suggested that the immunization way of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immu-nization could significantly enhance the humoral and cellular immunity of the bodies ,and provide the basis for further study on protective effect test in animals .
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Objective To determine vascular endothelial growth factor(VEGF)-460 gene polymorphism in Uyghurs and its relationship to urolithiasis in south Xinjiang. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),gene sequencing and genetic analysis methods were used in 200 urolithiasis patients of Uyghurs, and 200 healthy Uyghurs. Results The distribution of genotype and allele had no significant difference between urolithiasis patients and normal controls (P＞0. 05). The frequencies for the CC,TT and CT genotypes in patients with urolithiasis and normal controls were 1.5 %, 29.0 %, 69.5 % and 0. 5 %, 27.5 %, 72.0 %, respectively. The frequencies for C and T allele were 36.2%,63.7% and 36.9% ,63.1%, respectively. Conclusions The results of VEGF-460 gene polymorphisms indicate no significant relationship between patients with turolithiasis and normal controls in Uyghurs in south Xinjiang,which may not be urolithiasis susceptibility genetic locus.
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<p><b>OBJECTIVE</b>To study the effect of phosphorus on copper tolerance in Achyranthes bidentata.</p><p><b>METHOD</b>A PVC pipe experiment was conducted to study the interactive effects of phosphorus (P) and copper (Cu), on growth, elemental accumulation and chemical constituents of A. bidentata. Two levels of elemental P were applied at 0 (P0) and 100 ( P100) mg x kg(-1) soil with 5 levels of Cu at 0 (Cu0), 100 (Cu100), 200 (Cu200), 200 (Cu400), 200 (Cu600) mg x kg(-1) soil, respectively.</p><p><b>RESULT AND CONCLUSION</b>The biomass production between different Cu treatments, phosphorus treatment showed significant differences. The biomass reached the maximum value as the concentration of Cu and P was 100 mg x kg(-1). Low concentration of Cu improved the growth of A. bidentata. The growth was blocked as Cu concentration reached 200 mg x kg(-1) in soil, however the contents of oleanolic acid and ecdysterone in roots of A. bidentata had not influenced by Cu. P could improved the copper tolerance in A. bidentata and increased root yield. The Cu concentration in soil of the cultivation bases must be below 200 mg x kg(-1) in order to produce good quality of medicinal material.</p>