Objective To establish a stable cell line expressing transmembrane form of human blood group A antigen mimotope vaccine by transfecting malignant melanoma cell line B16, and to detect the cytotoxicity of the vaccine against melanoma cells. Methods Cultured B16 cells were classified into 4 groups, i.e.,P/F-M-pIRES group [transfected with the recombinant plasmid mimotope peptide/Fas-macrophage inflammatory protein (Mip)-pIRES], P/F-pIRES group (transfected with the recombinant plasmid mimotope peptide/FaspIRES), M-pIRES group (transfected with the recombinant plasmid Mip-pIRES), and pIRES group (transfected with the empty plasmid pIRES). B 16 cells were transfected through Lipofectamine 2000. Subsequently, RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of the mimotope peptide/Fas fusion gene and Mip3β in transfected B16 cells. Cell counting kit-8 (CCK-8) was used to evaluate the vaccinemediated complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC)against B16 cells. Results RT-PCR yielded specific DNA fragments with expected size. Western blotting revealed the anti-A antibody-binding activity of the recombinant mimotope peptide/Fas fusion protein. Factor analysis indicated significant differences in CDC (F = 244.522, P ＜ 0.01 ) and ADCC (F = 71.593, P ＜ 0.01 )against B16 cells between the 4 groups. Group comparisons demonstrated more intense CDC and ADCC in P/FM-pIRES and P/F-pIRES groups compared with M-plRES and pIRES groups, stronger ADCC in P/F-M-pIRES group in comparison with P/F-pIRES group (F = 15.42, P ＜ 0.05), but no significant difference in CDC was observed between M-pIRES and pIRES group. Conclusions The transmembrane form of human blood group A antigen mimotope vaccine could be stably expressed in B16 cells, and mediate ADCC and CDC against B16 cells in vitro.

Since the polyjuice potion ingredient is complex, we need to develop an analysis method with well separation and high stability to perform qualitative analysis. After dividing chemical components of Mahuang Decoction into fat-soluble and water-soluble constituents by gradient extraction, GC-MS was used to analyze the chemical components of the ethyl acetate extraction. The results showed that forty compounds had been identified by NIST MS search 2.0 standard mass spectrometry Library and literatures. Next, UPLC-Q-TOF-MS was applied to idendify the chemical components of the water extraction. The results showed that thirty-nine compounds had been identified by MZmine-2.9.1, Isotope Pattern, fragmentation regularity of mass spectrometry and literatures. This experiment will provide evidences for elucidation of the effective substance in Mahuang decoction and can be used as a simple, shortcut method for analysis and identification for the polyjuice potion.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods

ObjectiveTo observe the lung injury of rats caused by PM2.5 induced imbalance of TH17/Treg immune system and the intervention effect of two different TCM treatments.Methods Wistar male rats were randomly divided into normal group, model group, TCM-treated group1 and TCM-treated group 2. PM2.5-induced lung injury model was established by airway instillation. Model group was given normal saline for gavage. TCM- treated group 1 and 2 were given Yupingfeng Powder and Guomin Decoction combined with Zhisou Powder for gavage. The pathological changes of bronchial and lung tissues, the contents of IL-8, IL-10, IL-17, NE, and MUC5AC in serum and BALF were compared, and the expressions of Foxp3 and IL-17 in lung tissue of each group were analyzed.Results Compared with normal group, the contents of IL-8, IL-17, NE and MUC5AC in serum and BALF of model group increased significantly, while IL-10 decreased significantly (P<0.05,P<0.01); the expression of IL-17 increased significantly and the expression of Foxp3 decreased significantly in lung tissue (P<0.01). Compared with model group, the contents of IL-8, IL-17, and NE decreased in TCM-treated group 1 and 2, while the content of BALF IL-10 increased significantly (P<0.05). The content of IL-10 in serum increased significantly in TCM-treated group 2 (P<0.05); the protein expression of IL-17 of lung issue decreased significantly, and the protein expression of Foxp3 increased significantly (P<0.01). The pathological changes were improved significantly.Conclusion PM2.5 can induce lung injury caused by the imbalance of TH17/Treg. Both two treatments can significantly improve the lung injury induced by PM2.5 and the imbalance of TH17/Tregs immune system.

Objective To investigate the clinical application of fast locator in proximal femoral nail antirotation (PFNA) for treating femoral intertrochanter fractures.Methods A retrospective case series study was conducted on the clinical data of 188 patients with femoral intertrochanteric fractures who were treated with closed reduction and PFNA from October 2010 to October 2014.There were 86 males and 102 females,with an average age of 69.5 years (range,55-96 years).Before operation,the patients were divided into two groups according to their will.Group A (98 cases) used the self-made locator to identify the entry point,and Group B (90 cases) used the routine incision operation.The operative preparation time,operation time,fluoroscopy frequency,incision length of major nail,intraoperative blood loss,Harris scores of hip joint function at 4 months after operation,and complications were recorded and compared between the two groups.Results Group A had a longer operative preparation time than Group B[(10.6 ±2.4) minutes vs.(2.5 ±0.9) minutes].However,the operation time [(26.0 ±5.5) minutes vs.(56.6 ± 9.4) minutes],total operation time [(36.6 ± 6.6) minutes vs.(59.6 ± 9.9) minutes],fluoroscopy frequency [(18.9 ± 5.1) times vs.(31.2 ± 9.1) times],blood loss [(50.5 ± 3.7) ml vs.(220.5 ± 2.7) ml] and incision length of major nail [(2.5 ± O.7) cm vs.(5.5 ± 1.7) cm] in Group A were all less than those in Group B (P ＜ 0.05).There was no significant difference in the excellent and good rate of Harris score at 4 months after operation between the two groups (97％ vs.97％) (P ＞0.05).Postoperative incision infection,hypostatic pneumonia,and deep vein thrombosis occurred in some patients.No significant difference in the incidence rates of complications in two groups (14％ vs.10％) was found (P ＞ 0.05).Conclusion Femoral nail needle point locator can quickly identify the entry point,improve the nail placement accuracy and reduce surgical trauma as well as shorten operation time,and hence is worthy of clinical application in the treatment of femoral intertrochanteric fractures.

Objective To investigate the value of intravoxel incoherent motion MR imaging (IVIM-MRI)on monitoring impairment of renal function in hydronephrosis.Methods Left hydronephrosis model of Healthy New-Zealand rabbits made by ureteral incomplete obstruction were scanned by IVIM-MRI at 4, 8, 12, and 16 weeks.The change of IVIM-MRI quantitative parameters (ADC values, D values, D* values, and f values) in left hydronephrosis kidney were observed.The correlation between these parameters and glomerular filtration rate (GFR) were evaluated.Results ADC values, D values, D* values and f values at cortex and medulla in left hydronephrosis model of rabbits decreased gradually in the progression of renal damage after 4, 8, 12, and 16 weeks.For D value, f value and ADC values, there was statistical significant difference among the group of 12 weeks, the group of 16 weeks, the control group, and the group of 4 weeks;statistical significant difference was also observed between the group of 8 weeks and the group of 16 weeks by LSD contrast.ADC values, D values, D* values, and f values were positively correlated to GFR at 4, 8, 12, and 16 weeks for left hydronephrosis model of rabbits.Conclusion IVIM-MRI could monitor impairment of renal function in hydronephrosis dynamically.

Objective To observe the effects of scutellaria barbata extract (ESB) on proliferation, apoptosis and telomerase activity of human malignant glioma U251 cells in vitro.Methods Different concentrations of ESB (50, 25, 12.5, 6.25, 3.125 and 1.5625 mg/mL) were added into the medium cultured human malignant glioma U251 cells for 24, 48 and 72 h, respectively. And blank control group was also established. MTT assay was employed to detect the proliferation of U251cells. AnnexinV/PI staining and low cytometry (FCM) were used to detect the changes of apoptotic rate.And the telomerase activity of the cells was observed under the examination of telomeric repeat amplification protocol-PCR (TRAP-PCR)-ELISA.Results ESB inhibited the proliferation and induced the apoptosis of U251 cells. Interaction effect was found between the concentration of ESB and the treatment time of ESB by MTT assay (F=59.908, P=0.000); 50 mg/mL ESB for 72 h could most significantly inhibit the proliferation of U251 cells. Interaction effect was found between the concentration of ESB and the treatment time of ESB by AnnexinV/PI staining (F=6.548, P=0.000); 25mg/mL ESB for 72 h could most significantly induce the apoptosis of U251 cells. Interaction effect was also found between the concentration of ESB and the treatment time of ESB by TRAP-PCR-ELISA(F=138.433, P=0.000); the telomerase activity of the cells was the lowest by treatment with 50 mg/mL ESB for 72 h; negative correlation was noted between the telomerase activity of the cells and the apoptosis rate (r=-0.785, P=0.037); so as the telomerase activity of the cells and the inhibition effect (r=-0.278, P=0.042) Conclusion ESB may inhibit the proliferation and induces the apoptosis of U251 cells through down-regulating the telomerase activity.

Objective: To understand hepatitis B virus(HBV) infection in high school students in Guangxi, and to provide scientific foundation for hepatitis B prevention and control strategies. Methods: A selfdesigned epidemiological questionnaire was used among 2 632 participants. Information regarding general demographic characteristics, lifestyle habits, HBV infection history, knowledge of viral hepatitis prevention and control, as well as hepatitis B vaccination. Univariate and multivariate logistics regression analysis was used to analyze associated factors for hepatitis B infection. Results: A total of 192 out of 2 632 participants were found HBsAg positive and 2 440 were HBsAg negative. Multivariate analysis showed that family liver disease history and hairdressing injury positively associated with hepatitis B virus infection risk (OR=3.62, 95% CI=2.28-5.73;OR=3.06, 95%CI=1.94-4.83), and hepatitis B vaccination was negatively associated with hepatitis B virus infection risk (OR=0.08, 95%CI=0.05-0.11). Conclusion Family history of liver disease, hairdressing injury experiences, as well as low rate of hepatitis B vaccination may associate with relatively high prevalence of hepatitis B virus infection among high school students in Guangxi.

OBJECTIVETo develop an HPLC method for the determination of serum level of Crebanine (Cre) and study on the pharmacokinetics of Cre injection in rabbits.

METHODTo sample blood serum from the rabbits' ears which were injected the Cre by 2.0 mg x kg(-1) at different time and use HPLC to determine the concentration of Cre in it, the pharmacokinetic parameters were accessed by the DAS software.

RESULTCre was fitted to a two compartment open pharmacokinetic model in rabbits. There was no signifiant difference between the male and female rabbits'pharmacokinetic by t-test. The mainly pharmacokinetic parameters were: t1/2alpha = (3. 246 +/-0.222) min, t1/2beta = (36.67+/-5.52) min, Cmax = (1.401 +/- 0.062) mg x L(-1), Vd = (5.928 +/- 0.877) L x kg(-1), Cl = (0. 051 +/-0.003) L x min(-1) x kg(-1).

CONCLUSIONThis experiment can objectively show the pharmacokinetics regularity of Crebanine injection in rabbits. Crebanine injection was a speeding disposition drug (t1/2 <1 h) and disposed extensively and rapidly in rabbits.

Animals ; Aporphines ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Female ; Injections ; Male ; Metabolic Clearance Rate ; Plants, Medicinal ; chemistry ; Rabbits ; Stephania ; chemistry

OBJECTIVETo study the expression ratio of AML1-ETO9a (AE9a) isoform in t(8;21) acute myeloid leukemia (AML) and its clinical significance.

METHODSBone marrow samples from 44 newly diagnosed t(8;21) AML patients co-expressed AE9a and AE were screened by RT-PCR. The alteration of the AE9a expression ratio was monitored during follow-up by using quantitative real-time RT-PCR (qPCR).

RESULTSThe expression level of AE9a was markedly lower than that of AE in these patients. There was a positive correlation between the expression level of AE9a and AE in most of bone marrow samples. The transcript level of both AE9a and AE was decreased in the 44 patients after one course of standard chemotherapy, but the percentage of AE9a expression level was increased in comparison with that before treatment (P < 0.05). After one course of standard chemotherapy treatment, the percentage of AE9a in incomplete remission (ICR) patients was significantly higher than that in CR patients (P < 0.05). Relapsed patients had a higher AE9a ratio than the unrelapsed patients (P < 0.05). During the remission, the percentage of AE9a in 11/17 relapsed patients obviously elevated even while the expression of AE fusion gene at low level.

CONCLUSIONSAE9a and AE co-expressed in most of AML patients with t(8;21) translocation. The expression level of AE9a was lower than that of AE, and there is a positive correlation between the expression level of these two isoforms. The sensitivity of AE9a gene to the standard chemotherapy is less than that of the AE fusion gene. Monitoring the AE9a to AE ratio during the CR can predict the early relapse of the disease compared to monitoring the AE alone.

Adolescent ; Adult ; Child ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Gene Expression ; Humans ; Leukemia, Myeloid, Acute ; genetics ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Protein Isoforms ; genetics ; RUNX1 Translocation Partner 1 Protein ; Translocation, Genetic ; Young Adult

Objective:To summarize the risk factors, clinical manifestations, diagnosis and treatment of limb thrombosis in neonates.Methods:The clinical data of 14 neonates with limb thrombosis were hospitalized in neonatology department at Shengjing Hospital of China Medical University from February 2012 to February 2022 were retrospectively analyzed.Results:All the 14 cases of limb thrombosis were premature infants, with an average gestational age of 29 weeks and 5 days(27 weeks and 3 days to 33 weeks and 1 day), including eight cases of arterial embolism and six cases of venous embolism.Among them, 13(92.9%) cases were diagnosed with infectious diseases such as septicemia or neonatal necrotizing enterocolitis within 48 hours before embolization, and all had a history of peripheral arterial and venous catheterization.During the early stage of embolization, limb artery embolism was characterized by weakened distal artery pulsation, pale skin, gradual cyanosis and even gangrene.Limb venous embolism was manifested as limb swelling, skin congestion and cyanosis, but the arterial pulsation was normal.Fourteen cases were confirmed by vascular ultrasound.All the eight cases with arterial embolization were treated with heparin anticoagulation, five of which were cured, with an effective rate of 62.5%.The average time of heparin use in five cases was 2.5 days(2-3 days). One patient was not effective after 2 days of heparin treatment, and recovered after thrombectomy.Another two cases had distal limb gangrene, and them were treated with heparin for 5 days and 7 days.All of the six cases with venous embolism were cured, of which four cases were treated with heparin for an average of 8.5 days(4-15 days), and the other two cases were cured after general treatment.There were no bleeding events in the 12 infants treated with heparin.Conclusion:Peripheral arterial and venous catheterization during infection of preterm infants is the most common cause of limb thrombosis.The smaller body weight and gestational age, the thinner blood vessels, the higher risk of occurrence.Vascular ultrasound is the most commonly used examination method for neonatal thrombosis, and heparin anticoagulant therapy is the most commonly used treatment measure.When the treatment effect of heparin is not good, other treatment methods should be sought.