Aim To identify the molecular target of gabapentin in the treatment of postherpetic neuralgia(PHN). Methods The molecular target of gabapentin for PHN was analyzed by network pharmacology and molecular docking and confirmed by coprecipitation test. Rats were randomly divided into control group, model group, model+50 mg·kg-1 gabapentin group, model+100 mg·kg-1 gabapentin group, and model+200 mg·kg-1 gabapentin group, with nine rats in each group. The pain-related behaviors of the rats were measured at different time points. The mRNA and protein expressions of CACNA2D1, Bax, and Bcl-2 in rat spinal cord were determined by immunofluorescence, Western blot, and qPCR. Results CACNA2D1 was the target gene of gabapentin that determined via network pharmacology, molecular docking, and co-precipitation tests. After modeling, mechanical pain threshold and thermal pain threshold significantly decreased, and the number of apoptotic GABA cells significantly increased. However, after intraperitoneal injection of 50, 100, and 200 mg·kg-1 gabapentin, mechanical pain threshold and thermal pain threshold significantly increased(P<0.05), and the number of apoptotic GABA cells significantly decreased(P<0.01). Immunofluorescence and Western blot results showed that compared with the model group, with the increase of gabapentin concentration, the positive expression rate of Bax significantly decreased, and the positive expression rate of Bcl-2 and CACNA2D1 significantly increased. The mRNA expression levels of Bax, Bcl-2 and CACNA2D1 detected by qPCR were consistent with the results of immunofluorescence and Western blot. Conclusions Gabapentin up-regulates the expression of target protein CACNA2D1, inhibits the proapoptotic protein Bax, and promotes the expression of apoptotic inhibitor Bcl-2.

Objective To investigate the predictive value of quantitative flow ratio(QFR)on the occurrence of major adverse cardiovascular events(MACE)3 years after percutaneous coronary intervention(PCI)in patients with non-acute myocardial infarction.Methods This study included 139 patients with non-acute myocardial infarction who underwent PCI from January 2020 to June 2020 in the cardiac catheterization room of our hospital,all of them underwent post-PCI target vessel QFR measurements,and the incidence of MACE was followed up 3 years after PCI.The cut-off value of QFR was calculated by receiver operating characteristic(ROC)curve,according to which patients were divided into QFR＞0.95 group and QFR≤0.95 group,and patients were divided into MACE group and non-MACE group depending on whether MACE occurs.The independent influencing factors of post-PCI QFR in patients with non-acute myocardial infarction were investigated by univariate and multifactorial linear regression analysis.Univariate and multivariate Cox regression analysis was used to investigate the predictive value of post-PCI QFR in the occurrence of MACE 3 years after PCI in patients with non-acute myocardial infarction.The long-term prognosis of patients with QFR＞0.95 and those with QFR≤0.95 was compared by drawing the survival curve of no-MACE event.Results ROC curve analysis showed that post-PCI QFR predicted the occurrence of MACE 3 years after surgery in non-acute myocardial infarction patients with statistical significance(AUC 0.666,95％CI 0.556-0.777,P=0.003),and the cut-off value of MACE was 0.95.The sensitivity and specificity were 75.00％and 51.30％for the diagnosis of MACE with QFR≤0.95.Patients were divided into QFR≤0.95 group(n=74)and QFR＞0.95 group(n=65)according to the cut-off value.Multivariate linear regression analysis showed that the maximum area stenosis rate after PCI was an independent factor for post-PCI QFR(P＜0.001).Multivariate Cox regression analysis showed that body mass index(BMI),post-PCI QFR,three-vessel lesions and post-PCI QFR groups were independent influencing factors of no-MACE lifetime.The no-MACE survival curve showed that the long-term prognosis of patients in the QFR＞0.95 group was significantly better than that in the QFR≤0.95 group(x2=5.272,P=0.022).Conclusions The optimal cut-off value of post-PCI QFR for predicting the occurrence of MACE 3 years after PCI in non-acute myocardial infarction patients was 0.95,and patients with QFR≤0.95 had worse long-term prognosis,and BMI,three-vessel lesions,and post-PCI QFR≤0.95 were independent risk factors for the occurrence of MACE 3 years in non-acute myocardial infarction patients after PCI.

OBJECTIVE: To investigate the clinical features, accompanying gene mutation characteristics and prognostic factors of adult patients with acute myeloid leukemia with mutated NPM1 (NPM1AML). METHODS: Seventy-three patients with newly diagnosed adult NPM1AML were selected. The mutations of 22 genes were detected by second generation sequencing and 43 fusion genes of AML were detected by real-time fluorescent quantitative PCR. The Kaplan-Meier survival curve and Cox multivariate regression analysis were used to study the prognostic factors. RESULTS: A total of 74 NPM1 site mutations were detected in 73 patients with NPM1AML. The incidence rates were 92.0% L287fs, 2.7% Q289fs and W288fs, 1.4% L258fs and Q289H, among which 1 patient had 2 NPM1 mutations; the different mutation sites had no effect on the prognosis of NPM1AML. The median value of NPM1 variant allele frequency (VAF) was 35.4% (1.8%-56.6%). Based on the uppermost quartile of 38.4%, the patients were classified as NPM1 VAF>38.4% (NPM1AML) and NPM1 VAF≤38.4% (NPM1AML). Compared with NPM1AML, the early mortality rate was statistically significantly higher (33.3% vs 7.3%, P<0.05), and median EFS (148 d，95%CI 58-238 d vs 372 d，95%CI 264-480 d) (P<0.01) and median OS (179 d 95%CI 6-352 d vs 444 d) (P<0.01) were significantly shorter in NPM1 AML. A total of 126 accompanying gene mutation sites were detected in 87.7% of patients with NPM1AML. The patients with NRAS gene mutation displayed a higher rate of complete remission (100% vs 58%) (P<0.05) and longer median OS (not reached to 320 d, 95%CI 150-490 d) (P<0.05). The 43 fusion genes were examined in 65 out of 73 cases of NPM1AML, and in all the patients the fusion gene test was negative. Multivariate analysis showed that NPM1 VAF>38.4% was an independent prognostic factor for EFS (HR=3.1, 95% CI 1.6-6.4, P<0.01) and OS (HR=3.0, 95% CI 1.4-6.2, P<0.01). CONCLUSION The NPM1 gene mutation in AML patients often is accompanied by other gene mutations, while the coexistence of fusion genes is rare; high NPM1 mutant allele burden is an independent prognostic factor for adult AML patients with mutated NPM1.
Alleles ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; fms-Like Tyrosine Kinase 3

Objective To investigate the effect of cysteine protease inhibitor derived from S chistosoma japonicum(SjCys-tatin)on dextran sodium sulfate(DSS)-induced acute ulcerative colitis in mice.Methods Eighteen C57BL/6 mice were ran-domly divided into three groups:a control group treated with PBS(Group A),a DSS-induced-colitis group treated with PBS(Group B),and a DSS-induced-colitis group treated with SjCystatin(Group C).Colitis was induced in mice by giving 3％DSS orally for 7 days.During this period,the mice were daily injected with 10μg of SjCystatin or PBS only as a control intraperitone-ally.The mice were monitored daily for their clinical manifestations and given scores based on disease activity index(DAI).The severity of colonic inflammation was monitored by the macroscopic score and pathological change.The cytokine profile including TNF-α,IL-4,IL-6 and IL-10 in the supernatants of colon homogenate was detected by ELISA.Results Compared with Group A(0.50 ± 0.28),the DAI score increased significantly in Group B(9.30 ± 1.30)(F=86.86,P<0.01),with remarkable path-ological damages seen in colon tissues.and the levels of TNF-α and IL-6 were(321.33±67.01)and(403.58 ±180.51)pg/mL.The DAI score significantly reduced in Group C(6.67±1.57)as compared to Group B(F=86.86,P<0.01),with improve-ments in the macroscopic and microscopic pathology in mouse colon specimens.As compared to Group B,the levels of TNF-α [(188.14 ± 40.14)pg/mL] and IL-6 [(209.71 ± 48.47)pg/mL] significantly decreased(F=17.46 and 9.89,both P<0.01).Con-clusion SjCystatin has a significantly inhibitory effect for alleviating DSS-induced acute ulcerative colitis in C57BL/6 mice.

Objective To evaluate the accuracy of CT imaging of inferior vena cava and hepatic vein in patients with hepatic cystic echinococcosis. Methods A total of 100 patients with hepatic cystic echinococcosis were given triple-phase abdominal CT scan, and the reconstructed images of the inferior vena cava and hepatic vein were obtained and compared to the real situation during the surgical operation. The reliability of the CT evaluation of the echinococcus cyst lesion invading inferior vena cava and hepatic vein was analyzed. Results The compression displacement, half-globular, no clear demarcation between the lesion and blood vessel and narrowing of the vessel detected by CT evaluation were in accordance with the real situation under the surgical sight. However, four cases of clear demarcation between the lesion and blood vessel were affirmed wrong under the surgical sight. There were three cases of over diagnosis of vascular stenosis judgments. Conclusion The CT images of hepatic vein and inferior vena cava can clearly show the relationship between the hepatic echinococcus cyst and hepatic vein and inferior vena cava.

The agronomic traits (plant height, root diameter, root length, first lateral root height, lateral root amount, root weight) of 18 Polygala tenuifolia samples with different agronomic traits were analyzed, respectively. HPLC was used to analyze three main characteristic components including tenuifolin, polygalaxanthone Ⅲ, and 3,6'-disinapoyl sucrose. At last, the correlation between six agronomic traits and three main characteristic components were analyzed by scatter plot. We found no significant correlation between root diameter and three main characteristic components. There were no obvious correlations between tenuifolin and the remaining five agronomic traits. Short root length and first lateral root height as well as high lateral root amount resulted in high levels of polygalaxanthone Ⅲ in P. tenuifolia samples. High levels of 3,6'-disinapoyl sucrose were observed in P. tenuifolia samples with longer root. So, the current commodity criteria and traditional breeding of P. tenuifolia did not conform to pharmacopoeia standards, which excellent medicinal materials should have high contents of the main characteristic components. It was urgent to revise the current commodity criteria and breeding methods.

OBJECTIVETo establish the primary myelofibrosis (PMF)-induced pluripotent stem cell line (iPSC) by means of iPSC techinique so as to provide a experimental model for studying the blood disease mechanisms.

METHODSInduced pluripotent stem cells were generated from mononuclear cells isolated from a PMF patient with JAK2(V617F) mutation by using episomal vectors.

RESULTSPMF-derived iPSC was established from the patient with JAK2(V617F) gene mutation. The PMF-iPSC could be stably passaged, highly expressed pluripotent genes as human embryonic stem (ES) cells, and were able to form teratoma in NOD/SCID mice in vivo. H & E staining of the teratoma showed the presence of tissue type derived from all three embryonic germ layers. Sanger sequencing confirmed that PMF-derived iPSC carried different allele burdens of JAK2(V617F) gene mutation.

CONCLUSIONThe interation-free iPSC from primary myelofibrosis patient in vitro has been established. This PMF-derived iPSC line provides a valuable tool for studying the pathogenesis, screening of chimical drugs and realizing the standard therapy of PMF.

Alleles ; Animals ; Cell Culture Techniques ; Humans ; Induced Pluripotent Stem Cells ; Janus Kinase 2 ; genetics ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Mutation ; Primary Myelofibrosis

The study was aimed to investigate the effect of anti-mouse CD122 antibody on the hematopoietic repopulating capacity of cord blood CD34⁺ cells in a humanized murine model-non obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. After sublethal irradiation with γ-ray, NOD/SCID mice were intraperitoneally injected with 200 µg mouse isotype control antibody or anti-mouse CD122 antibody. Human cord blood CD34⁺ cells or phosphate-buffered saline (PBS) were injected via the tail vein at 6-8 hours later. Cohort of the mice injected with anti-mice CD122 antibody or control antibody alone were sacrificed at different time point (at week 2, 3, and 4 weeks) after the injection, and the percentage of NK cells in the peripheral blood was analyzed by flow cytometry. To evaluate the effect of anti-mouse CD122 antibody on the repopulating capacity of cord blood CD34⁺ cells in the recipient mice, phenotype analysis was performed in the bone marrow at 6 and 8 weeks after the transplantation. The results showed that the proportion of NK cells in the peripheral blood were (4.6 ± 0.6)% and (5.7 ± 1.7)% at week 2 and 3 after anti-CD122 antibody injection respectively,which decreased by 60%, compared with the mice injected with isotype control antibody. After 6 and 8 weeks of cord blood CD34⁺ cell transplantation,the percentage of human CD45⁺ in the bone marrow of the recipient mice treated with anti-mice CD122 antibody was (63.0 ± 12.2)% and (53.2 ± 16.3)%,respectively,which were dramatically higher than that in the mice treated with isotype control antibody (7.7 ± 3.6)% and (6.1 ± 2.4)%. Moreover,at 8 weeks after transplantation,human CD34⁺ cells appeared significantly in the recipients treated with anti-CD122 antibody. It is concluded that the anti-mouse CD122 antibody enhances the hematopoietic repopulating capacity of cord blood CD34⁺ cells in the NOD/SCID mice through decreasing the proportion of NK cells.
Animals ; Antibodies ; immunology ; Antigens, CD34 ; Bone Marrow ; Cord Blood Stem Cell Transplantation ; Fetal Blood ; immunology ; Hematopoietic Stem Cell Transplantation ; Hematopoietic System ; cytology ; immunology ; Humans ; Interleukin-2 Receptor beta Subunit ; immunology ; Killer Cells, Natural ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Transplantation, Heterologous

Objective To screen mutations in genes including ASXL1, TET2, IDH1, IDH2, SETBP1, MPL515, JAK2 exon 12 and JAK2V617 in 135 polycythemia vera (PV) patients. To assess progreasson and genomics characteristics post polycythemic myelofibrosis. Methods DNA sequencing of ASXL1(Exon12),TET2 (Exons 3-11),IDH1(Exon4),IDH2(Ex-on4),SEPBP1(Exon4),JAK2 exon 12 and MPL515 (Exon 10) genes were carried out using Sanger method. JAK2V617 muta-tion was detected by allele-specific PCR in patients with PV. In the mean time, the mutation load of JAK2V617F allele (V617F%) was evaluated by real-time PCR using Tagman MGB probe. Then, the significant of gene mutations and clinical outcomes of post-PV Myelofibrosis(PPMF)was analyzed. To study risk factors of PPMF, logistic regression were employed. Results ASXL1, TET2, IDH1, IDH2 were mutated in 7.69%(8/104), 5.26%(1/19) , 0.08%(1/120) and 0.08%(1/121) of all PV patient respectively. JAK2 was mutated in 82.22%(111/135) of PV patients with mutation rate of exon12 of 2.96%(4/135) and there were no mutation of MPL515 and SETBP1 in PV patients. ASXL1 mutation was found in 31.82%(7/22) PPMF patients. Spearman analysis showed that ASXL1 is correlated with JAK2V617F (V617F%) (rs=0.298,P=0.002). The hemo-globin was lower in patients with ASXL1 mutation than patient without mutation (wild type). Leukocyte count, V617F%>50%rate, thrombosis and PPMF were higher in patients with ASXL1 mutation than that of ASXL1 wild type(P＜0.05). ASXL1 mu-tation, V617F%>50% rate and splenomegaly were all risk factors of PPMF. Conclusion ASXL1 mutation is the risk-fac-tor of PPMF and may promote V617F%by some mechanism.

OBJECTIVETo evaluate the application of side-to-side anastomosis of the lesser curvature of stomach and jejunum in laparoscopic Roux-en-Y gastric bypass (LRYGB).

METHODSClinical data of 29 patients with type 2 diabetes mellitus (T2DM) undergoing side to side anastomosis of the lesser curvature of stomach and jejunum in LRYGB from May 2012 to November 2012 in Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University were analyzed retrospectively.

RESULTSAll the procedures were successfully completed without conversion to laparotomy. The side-to-side anastomosis of the lesser curvature of stomach and jejunum avoided the laparoscopic suture. No gastrojejunostomy anastomotic bleeding, fistula, obstruction and other complications occurred after operation and no complications of gastrojejunostomy anastomosis were found during a follow up of 1 to 7 months.

CONCLUSIONSSide-to-side anastomosis of the lesser curvature of stomach and jejunum in LRYGB can manipulate the size of anastomosis accurately and avoid the laparoscopic suturing. It is simple and easy to learn.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Follow-Up Studies ; Gastric Bypass ; methods ; Humans ; Jejunum ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach ; surgery ; Treatment Outcome