The methylation of CpG island in promoter region of E-cad gene which usually leads to inactivation of E-cad gene is closely related to the occurrence, development, invasion, metastasis and prognosis of a variety of gastrointestinal cancer, such as esophageal carcinoma, gastric carcinoma, hepatocellular carcinoma,cholangiocellular carcinoma, carcinoma of gallbladder, pancreatic carcinoma and colorectal cancer. Further study would probably guide the early diagnosis, gene therapy, prophylaxis and predicting recurrence of tumor metastasis of gastrointestinal cancer.

Objective To study the changes of CDH1 gene promoter CpG island methylation and its clinical significance in patients with esophagus and stomach double primary carcinoma(ESDC).Methods The expression of CDH1 gene methylation in cancerous tissues and adjacent cancerous tissues in 18 cases of ESDC were detected using methylation-specific PCR method. Results Eighteen patients were endoscopically diagnosed as ESDC between Jan. 2007 and Sep. 2009 in the 4th Hospital Affiliated to Hebei Medical University. The positive methylation of CDH1 gene in tissues of esophageal squamous cell carcinoma (ESCC)and adjacent cancer were 66.7% and 33. 3%, respectively, with significant difference (χ2= 4. 167, P = 0. 031). Whereas the positive methylation of CDH1 gene in tissues of gastric carcinoma (GA) and adjacent cancer were 77.8% and 44.4%, respectively, without statistical difference (χ2=1.786, P= 0. 180). There was no significant difference (P=0. 500) in positive rate of CDH1 gene methylation between ESCC tissues and GA tissues in same individual with ESDC. For 18 patients with ESDC, consistent change of CDH1 methylation in tissues of two kinds of cancers was found in 16 patients with a total agreement of 88.9 % (positive agreement of 66.7 % and negative agreement of 22. 2%). Statistical analysis showed a significant correlation between two groups (P = 0. 005). Conclusion In patients with ESDC, there is a high consistency of CDH1 methylation change, between ESCC and GA,which suggests that two kinds of cancer may have similar risk factors and molecular mechanisms.

To investigate the role of CD4+, CD25+, and CD127low regulatory T cells (Tregs) in multiple myeloma (MM). Methods:Levels of CD4+T cells and Tregs, as well as expression of CTLA-4 and apoptosis-related proteins, such as CD95, bcl-2, and Caspase3 of Tregs in peripheral blood of 30 patients with newly diagnosed cases, 27 patients under of complete remission (CR) from multiple myeloma patients, and 25 healthy adults were analyzed by flow cytometry. Results:The percentage of CD4+T cells in the untreated group was significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was significantly higher than that of the CR group and control group (P<0.05), which in ISSⅢpatients of the untreated group was significantly higher than that in I/II(P<0.05). No significant difference of CD95 expression in Tregs was observed among the three groups. The expression of CTLA-4 in Tregs from the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls (P<0.05). The expression of bcl-2 in Tregs in the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls(P<0.05). The expression of Caspase3 in Tregs from the untreated group and CR group were all significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was positively correlated with the proportion of bone marrow plasma cells (P<0.05). The percentage of Tregs in CD4+T cells from 15 MM patients who received bortezamib and dexamethasone (VD) chemotherapy was negatively correlated to the ratio of plasma cell reduction after the first VD chemotherapy (r=0.735, P<0.01). Conclusion:The level of Tregs in the peripheral blood of MM patients was positively correlated with tumor burden and progression of disease, but was negatively correlated with curative effect. The increased level of Tregs was associated with their strengthened anti-apoptosis function.

Objective:To investigate the clinical effect and safety of reduction and fixation for the anterior glenoid fracture through an axillary approach.Methods:Two autopsy specimens (a total of 4 shoulder joints) were used to simulate the surgery of open reduction and internal fixation to treat anterior glenoid fracture through an axillary approach. Specimens were placed in the lateral decubitus position. An incision was made in the posterior axillary line to expose the lateral side of the scapula through the interval ahead of the latissimus dorsi muscle. Attended to the separation and protection of the axillary nerve, posterior humerus artery, thoracic dorsal nerve, and scapular artery. The surgical area of the axillary approach was divided into the upper "quadrilateral area" and the lower "trilateral area", which exposed the anteroinferior glenoid, neck, and the full length of lateral border of the scapula. Thirteen cases (7 males and 6 females) were involved in this study, all patients were diagnosed with anterior glenoid fracture and treated by open reduction and internal fixation through the axillary approach between April 2018 and December 2020. Constant-Murley score and Disabilities of the Arm, Shoulder and Hand (DASH) were used to evaluate the clinical efficacy.Results:Thirteen patients were enrolled for final analysis, which included 12 right cases and 1 left case. The average age was 50.38±13.74 years (range 24-67 years). All 13 patients were classified as Ideberg type Ia fracture. Anatomical buttress plates were used in 5 cases, cannulated screws combined with metacarpal plates in 7 cases, and distal radius plate in 1 case. The average length of follow-up was 13.00±5.97 months (range 6-26 months). No delayed union or malunion in all cases at the last follow-up visits. The average of Constant-Murley score was 62.46±10.26 points (range 45-83) and the DASH score was 27.56±9.76 points (range 14.14-43.33) at the three months follow up visits. At 6 months postoperatively, the Constant-Murley score was 80.85±8.32 points (range 65 to 90) and the DASH score was 11.47±8.56 points (range 0 to 35.00). Constant-Murley score at the final patient's follow-up visit was 84.54±8.95 points (range 70-95), and the DASH score was 10.94±8.67 points (range 1.67 to 33.33 points). The joint function gradually recovered with time. The functional scores at 3 months, 6 months after surgery, and final follow-up visit had significant differences ( P<0.05). Conclusion:The axillary approach is a safe and feasible approach for the treatment of the anterior glenoid fracture. This approach exposes the anteroinferior glenoid and the lateral border of the scapula, which reduces rotator-cuff injury and achieves anatomic reduction and strong fixation of the fracture. The axillary approach surgery also allows early functional physiotherapy after surgery.

OBJECTIVETo investigate the changes of relative telomere length (RTL) of peripheral blood (PB) CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺T lymphocytes, CD19⁺B lymphocytes and bone marrow (BM) CD34⁺ cells and its association with disease severity in untreated patients with immuno-related pancytopenia (IRP).

METHODSThe PB CD3⁺ , CD3⁺ CD4⁺ , CD3⁺ CD8⁺ T lymphocytes, CD19⁺ B lymphocytes, and BM CD34⁺ cells were purified by magnetic activated cell sorting (MACS), and RTL were measured with flow-fluorescence in situ hybridization (FLOW-FISH).

RESULTSThe RTL of CD3⁺, CD3⁺CD4⁺ , and CD3⁺CD8⁺T lymphocytes in untreated IRP patients were (27.754 ± 16.323)%, (7.526 ± 3.745)% and (25.854 ± 14.789)%, respectivly, which were significantly shorter than those in healthy-controls (54.555 ± 19.782)%, (12.096 ± 2.805)%, and (38.367 ± 4.626)% (P<0.05). The RTL of CD19⁺ lymphocytes in untreated IRP patients was (22.136 ± 16.142)%, which was significantly shorter than that in healthy controls (42.846 ± 16.353)% (P<0.01). There was no significant difference of BM CD34⁺ cells RTL between the untreated IRP patients (22.528 ± 21.601)% and the healthy controls (23.936 ± 19.822)% (P>0.05). There were significantly positive correlations between the RTL of B lymphocytes and the count of white blood cell (r=0.706, P=0.015). There were negative correlations between RTL of B lymphocytes and the clinical symptoms (r=-0.613, P=0.045) and positive correlations with therapeutic effect (r=0.775, P=0.005).

CONCLUSIONThe shorter RTL of CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD19⁺ lymphocytes, and the normal RTL of BM CD34⁺ cells in untreated IRP patients were identified, which might imply that IRP is a type of acquired autoimmune diseases.

Adolescent ; Adult ; B-Lymphocyte Subsets ; immunology ; Child ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; T-Lymphocyte Subsets ; immunology ; Telomere ; ultrastructure ; Young Adult

OBJECTIVETo explore the pathogenesis of abnormal WT1 expression in paroxysmal nocturnal hemoglobinuria (PNH).

METHODSThe expression of WT1 mRNA in CD59⁻ and CD59⁺ bone marrow mononuclear cells (BMMNC) were measured by semi-quantitative reverse transcription PCR. After WT1 gene silence by RNA interference (RNAi) technology, biological characteristics of BMMNC were investigated by flow cytometry.

RESULTSThe relative expression of WT1 mRNA in PNH CD59⁻ BMMNC (1.06 ± 0.12) was significantly higher than that in PNH CD59⁺ BMMNC (0.90 ± 0.12) and normal BMMNC (0.86 ± 0.05, P<0.05), but there was no significant difference between PNH CD59⁺ BMMNC and normal BMMNC (P>0.05). WT1 mRNA expression in PNH was positively correlated with the proportion of CD59⁻ cells (r²=0.490, P=0.016), but had no relationship with the proportion of CD59⁺ cells. After WT1 gene silence by siRNA in PNH CD59⁻ BMMNC, WT1 mRNA expression was decreased. The proportions of G0/G1 phase in PNH CD59⁻ cell blank control group and siRNA-scr transfected group were (92.73 ± 3.71)% and (93.06 ± 4.14)%, and the proportions of S phase were (6.99 ± 3.61)% and (6.73 ± 4.08)%, respectively. The proportions of G0/G1 and S phase in siRNA-WT1 transfected group was (94.46 ± 3.71)% and (5.40 ± 3.55)%, respectively. There were significant differences in the proportions of G0/G1 phase and S phase among the controls, siRNA-WT1 transfected group and siRNA-scr transfected group (P<0.05). The rate of apoptosis in siRNA-WT1 transfected group [(35.91 ± 22.36)%] was significantly higher than those in controls [(26.12 ± 17.10)%] and siRNA-scr transfected group [(27.39 ± 18.99)%] (P<0.05).

CONCLUSIONsiRNA-WT1 could effectively suppress the WT1 gene expression of CD59⁻ clone in PNH patients, inhibit its proliferation, and promote its apoptosis. WT1 gene expression might contribute to PNH clone proliferation.

Adolescent ; Adult ; Aged ; Apoptosis ; Bone Marrow Cells ; metabolism ; Cell Cycle ; Female ; Hemoglobinuria, Paroxysmal ; metabolism ; pathology ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; RNA Interference ; RNA, Messenger ; genetics ; WT1 Proteins ; genetics ; metabolism ; Young Adult

OBJECTIVETo explore the expression levels of terminal complement complex (C5b-9) and CD62p on platelets and the soluble C5b-9 (sC5b-9) level in serum in patients with PNH or PNH-aplastic anemia (AA).

METHODSSerum levels of sC5b-9, complement C3 and C4 were detected by using ELISA in 25 patients with PNH/PNH-AA. The quantities of C5b-9 and CD62p on the membrane of platelets were detected by flow cytometry.

RESULTS①In PNH/PNH-AA group, the serum sC5b-9 level [390.27(265.73-676.87) μg/L] was lower than that in control group [540.39(344.20-1 576.78) μg/L] (P<0.01). ②The platelet PNH clone (CD59⁻CD61⁺/CD61⁺) size [50.58(23.29-81.60)%] was bigger in the PNH/PNH-AA group than that [23.57(15.58-29.02)%] in control group (P<0.01). The percentages of C5b-9 deposition (C5b-9⁺CD61⁺/CD61⁺) were higher on the PNH clone platelets (CD59⁻CD61⁺) in the PNH/PNH-AA group [(17.53 ± 6.27)%] than those on the normal platelets (CD59⁺CD61⁺) in PNH patients 11.33±5.03）%］ and control [(10.88±3.58)%] group (P<0.01). ③ The expression of CD62p (CD62p⁺CD61⁺/CD61⁺) on PNH clone platelets in PNH patients [(61.98 ± 11.71)%] was higher than that on the normal platelets in PNH patients [(43.76±11.30)%] and control group [(38.23±18.07)%] (P<0.01). In addition, the expression of CD62p on normal platelets was higher in PNH patients than control (P<0.05). ④The deposition of C5b-9 positively correlated with the expression of CD62p on the platelets (r=0.559, P=0.002).

CONCLUSIONDeficiency of CD59 antigen on platelets in PNH patients may lead to the deposition of C5b-9 on its membrane and its dysfunction, which may contribute to thrombosis events in PNH.

Anemia, Aplastic ; Blood Platelets ; Clone Cells ; Complement Membrane Attack Complex ; Flow Cytometry ; Hemoglobinuria, Paroxysmal ; Humans ; P-Selectin ; Thrombosis