Efficacy of prophylactic antibiotics for the prevention of neutropenic fever in patients with multiple myeloma receiving high-dose cyclophosphamide for stem cell mobilization.

High-dose cyclophosphamide (HD-Cy) (3 g/m2) plus granulocyte colony-stimulating factor (G-CSF) is a very effective regimen for peripheral blood stem cell (PBSC) mobilization. Unfortunately, it is associated with an increased risk of neutropenic fever (NF). We analyzed the effect of NF on PBSC apheresis results and the efficacy of prophylactic antibiotics for the prevention of NF associated with HD-Cy plus G-CSF for PBSC mobilization in patients with newly diagnosed multiple myeloma (MM). First, patients were divided into NF ( +) and NF ( -) groups according to whether they suffered from NF during mobilization. Second, we divided patients into an antibiotic prophylaxis group and a nonantibiotic prophylaxis group according to whether antibiotic prophylaxis was used during the mobilization period. Our study showed that NF( +) patients (n = 44) had lower CD34 + cell dose collection (median 2.60 versus 5.34 × 106/kg, P < 0.001) and slower neutrophil engraftment and platelet engraftment (median 11 versus 10 days, P = 0.002, and median 13 versus 11 days, P = 0.043, respectively) than NF( -) patients (n = 234). Of note, the nonantibiotic prophylaxis group patients (n = 30) had a 26.7% incidence of NF. In the patients receiving antibiotic prophylaxis (n = 227), the incidence was reduced to 9.3% (P = 0.01). The antibiotic prophylaxis patients had higher CD34 + cell collection (median 5.41 versus 2.27 × 106/kg, P < 0.001) and lower hospitalization cost of mobilization ($ median 3108.02 versus 3702.39, p = 0.012). Thus, our results demonstrate that NF is associated with lower CD34 + cell collection and that antibiotic prophylaxis can reduce the incidence of NF and improve stem cell mobilization and collection outcomes, which reduces the hospitalization cost of mobilization.

On-demand plerixafor added to high-dose cyclophosphamide and pegylated recombinant human granulocyte colony-stimulating factor in the mobilization of patients with multiple myeloma: a treatment with high effectiveness, convenient, and affordable cost.

Objective: The combination of high-dose cyclophosphamide (HD-Cy) (3g/m2) plus granulocyte colony-stimulating factor (G-CSF) and on-demand plerixafor (PXF) has been considered an effective mobilization regimen of patients with multiple myeloma(MM). However, the daily multi-injection regimen of G-CSF poses challenges. This study delves into the efficiency and cost implications of a novel approach, using HD-Cy alongside pegylated G-CSF (PEG G-CSF) and on-demand PXF. Unlike G-CSF, which necessitates daily injections, the half-life of PEG G-CSF extended allows for a single injection. Methods: A retrospective analysis was conducted on 350 MM patients, which were categorized based on their mobilization regimens: Cy+PEG G-CSF+/-PXF (n=66), Cy+PEG G-CSF (n=91), Cy+ G-CSF (n=169), and G-CSF+PXF (n=24). Results: Mobilization with Cy+PEG G-CSF+/-PXF(8.79)yielded a notably higher median CD34+ cell count compared to the other regimens: Cy+PEG G-CSF(4.96), Cy+G-CSF (4.65), and G-CSF+PXF (2.99) (P<0.001). The percentage of patients who achieved >6×106/kg CD34+ cells was significantly higher in the Cy+PEG G-CSF+/-PXF group (77.3%) than in the other mobilization regimens: Cy+PEG G-CSF (41.8%), Cy+ G-CSF (37.3%), and G-CSF+PXF (8.3%) (P<0.001). From a cost perspective, the Cy+PEG G-CSF+/-PXF approach was more economical than the G-CSF+PXF strategy but was marginally costlier than the other two methods. A multivariate assessment highlighted that the combination of Cy+PEG G-CSF with on-demand PXF had a superior potential to achieve the desired harvest (6×106/kg) compared to the Cy+PEG G-CSF protocol without PXF. The incremental cost-effectiveness ratio for each 1% increase in the probability of achieving a successful optimal harvest was $ 97.02 per patient. The incidence of neutropenic fever was 3.0% in the Cy+PEG G-CSF+/-PXF group. Conclusion: The combination of on-demand PXF with HD-Cy and PEG G-CSF offers a cost-effective approach with a high mobilization success rate, manageable side effects, and the convenience of fewer injections. It stands as a promising mobilization strategy for MM patients.

Lymphoepithelioma-like neoplasm of the biliary tract with 'probable low malignant potential'.

AIMS: Lymphoepithelioma-like carcinomas (LELCs) are uncommon epithelial cancers characteristically showing two distinct components consisting of malignant epithelial cells and prominent dense lymphoid infiltrate. Hepatic LELCs consist of two types, the lymphoepithelioma-like hepatocellular carcinoma and lymphoepithelioma-like cholangiocarcinoma (LEL-CCA), with the latter being strongly associated with Epstein-Barr virus (EBV). METHODS AND RESULTS: We present a series of three cases of intrahepatic biliary EBV-associated LEL tumours in which the biliary epithelial component showed a distinctly benign appearance, instead of the usual malignant epithelial features of a typical CCA or EBV-associated LEL-CCA. In the lesions, the biliary epithelium showed interconnecting glands or cords of cells. All had a very low proliferation (Ki-67) index. Immunohistochemistry for IDH1 and TP53 performed on two cases was negative and molecular tests for EGFR and KRAS gene mutations performed on one were negative. Prognosis was very good in all three cases, with patients alive with no evidence of disease 24-62 months after surgery. Intriguingly, all three cases had co-infection of HBV and EBV. These cases are also discussed in the context of the 63 cases of LEL-CCA available in the literature, with a focus on epidemiology, clinicopathological features and potential research interests. CONCLUSIONS: Based on the distinct clinicopathological features and unique survival benefits, we believe these tumours represent the benign end of the spectrum of EBV-associated lymphoepithelial biliary carcinomas. Whether these tumours require a revision of the current nomenclature to 'lymphoepithelioma-like neoplasm of the biliary tract with probable low malignant potential' will require more detailed analysis with larger case-series.

Nimesulide inhibits the growth of human esophageal carcinoma cells by inactivating the JAK2/STAT3 pathway.

Although selective COX-2 inhibitors have cancer-preventive effects and induce apoptosis, the mechanisms underlying these effects are not fully understood. This study investigated the effects of nimesulide, a selective COX-2 inhibitor, on apoptosis and on the JAK/STAT signaling pathway in Eca-109 human esophageal squamous carcinoma cells. The effects and mechanisms of nimesulide on Eca-109 cell growth were studied in culture and in nude mice with Eca-109 xenografts. Cells were cultured with or without nimesulide and/or the JAK2 inhibitor AG490. Cell proliferation was evaluated using the MTT assay, and apoptosis was investigated. COX-2 mRNA expression was measured using reverse transcription polymerase chain reaction, and protein expression was detected by Western blot analysis, immunohistochemistry, and flow cytometry. Nimesulide significantly inhibited Eca-109 cell viability in vitro in a dose- and time-dependent manner (P<0.05). Nimesulide also induced apoptosis, which was accompanied by a significant decrease in the expression of COX-2 and survivin and an increase in caspase-3 expression. Nimesulide downregulated the phosphorylation levels of JAK2 and STAT3, and JAK2 inhibition by AG490 significantly augmented both nimesulide-induced apoptosis and the downregulation of COX-2 and survivin (P<0.05). In vivo, nimesulide inhibited the growth of Eca-109 tumors and the expression of p-JAK2 and p-STAT3. Thus, nimesulide downregulates COX-2 and survivin expression and upregulates caspase-3 expression in Eca-109 cells, by inactivating the JAK2/STAT3 pathway. These effects may mediate nimesulide-induced apoptosis and growth inhibition in Eca-109 cells in vitro and in vivo.

Comparison of hand-sewn and stapled anastomoses in surgeries of gastrointestinal tumors based on clinical practice of China.

BACKGROUND: There is a lack of studies comparing stapled suturing and hand-sewn suturing in the surgeries of gastrointestinal tumors based on the clinical practice of Chinese surgeons. METHODS: Data were retrospectively collected from 499 patients who underwent surgery to remove gastrointestinal tumors from January 2008 to December 2009. The patients were divided into two groups according to the method of digestive tract reconstruction: 296 patients received stapled suturing and 203 patients received hand-sewn suturing. The operation time, postoperative hospital stay, postoperative recovery and complications of the patients were evaluated and compared between the two groups. RESULTS: The stapling procedure took shorter operative time compared to the hand-sewn procedure for gastric carcinoma, colorectal cancer and esophageal carcinoma (P < 0.05). There was no significant difference between the two groups in postoperative hospital stay (P > 0.05). Patients receiving stapled suturing also showed shorter recovery for gastric cancer, colorectal cancer, and shorter time to recovery of normal gastrocolorectal motility compared with patients in the hand-sewn group (P < 0.05). However, there was no difference between the two groups in terms of normal time to commencing liquid diet for esophageal cancer patients (P > 0.05). We also found that the stapled procedure showed a lower incidence of anastomotic leakage, anastomotic hemorrhage and stump leakage in treating colorectal cancer or gastric carcinoma compared with the hand-sewn procedure (P < 0.05). CONCLUSIONS: Application of the stapler in treating gastrointestinal tumors demonstrated better effects on patients in terms of surgical operation time, recovery time to normal functions, and occurrence of complications compared to hand-sewn anastomosis, especially in gastric carcinoma and colorectal cancer.

Prognostic effect of p53 expression in patients with completely resected colorectal cancer.

The p53 protein is closely involved in the carcinogenesis of many kinds of cancers. Though the prognostic role of p53 expression for the survival of colorectal cancer (CRC) patients has been preliminarily identified, the prognostic effect of p53 expression in patients with completely resected CRC is still unclear. Therefore, a retrospective cohort study was performed to assess the prognostic role of p53 expression for overall survival in patients with completely resected CRC. A total of 153 patients (mean age 50.9 years) with completely resected CRC was finally included in the retrospective cohort study. Kaplan-Meier product-limit methods and log-rank test were used to estimate overall survival distribution and test the difference. In addition, multivariable analysis by Cox regression model was also used to test the prognostic role of p53 expression on overall survival by adjusting for other confounding factors. Of those 153 CRC patients, 62 (40.5 %) were positive for p53 protein expression in the tumor tissues. The log-rank test showed that there was an obvious difference in the overall survival between the p53-positive group and the p53-negative group (P < 0.001). Multivariable analysis by Cox regression model further showed that p53 protein expression was an independent predictor of shorter overall survival in patients with completely resected CRC (hazard ratio [HR] = 1.77; 95 % confidence interval [95 % CI] 1.15-2.71, P = 0.009). Therefore, p53 protein expression in the tumor tissue is an independent predictor of shorter overall survival in patients with completely resected CRC.

[Observation of humoral immunity reconstitution and its relationship with infection after autologous hematopoietic stem cell transplantation for patients with multiple myeloma].

OBJECTIVE: To study the humoral immunity reconstitution and its relationship with infection in patients with multiple myeloma (MM) after undergoing autologous hematopoietic stem cell transplantation (auto-HSCT). METHODS: Forty-two MM patients undergoing auto-HSCT were included in this study. Peripheral blood were obtained for immunoglobulin detection, including IgG, IgA and IgM before transplantation and 1, 3, 6, 12, 18 and 24 months after transplantation. The time, type, pathogen of infection between 1 and 24 month after transplantation were analyzed. RESULTS: The level of IgA at 6 month [(0.75±0.59) g/L] after auto-HSCT was lower than that of pre-auto-HSCT [(1.04±0.70) g/L], and reached the level of pre-auto-HSCT at 9 months [(0.99±0.52) g/L] after auto-HSCT. The level of IgM reached the level of pre-auto-HSCT [(0.45±0.26) g/L] at 3 months after auto-ASCT [(0.50±0.26) g/L]. The level of IgG reached the level of pre-auto-HSCT [(9.80±2.98) g/L] at 1 month after auto-HSCT [(11.09±2.69) g/L], and higher than that of pre-auto-HSCT at 9 months after auto-HSCT [(12.07±3.57) g/L]. The level of IgG with IgG-type MM was higher than that of patients with light-chain type and IgD-type MM at 6, 9 and 12 months after auto-HSCT. The IgA level of patients who obtained complete remission (CR) is much higher than that of patients who obtained nCR in IgG-type patients. The incidence of infection in 6 month after auto-HSCT was higher than that of (6-12) month and >12 month after auto-HSCT. The incidence of infection was strongly negative correlated with IgA (r =-0.943, P=0.005) and IgG (r=-0.943, P=0.005) level. The frequency of viral infection was also negatively correlated with IgA and IgG. CONCLUSION: The reconstitution time of IgG, IgA and IgM was different in MM patients after auto-HSCT. IgG recovered first, then IgM, and IgM the last. The incidence of infection was negatively correlated with IgA and IgG. With the recovery of IgG and IgA, the incidence of infection was decreased accordingly.

[Clinical significance of abnormal protein bands in multiple myeloma treated with bortezomib-based induction regimen and autologous stem cell transplantation].

OBJECTIVE: To study the clinical significance of abnormal protein bands (APB) in multiple myeloma (MM) patients treated with bortezomib-based induction regimen and autologous stem cell transplantation (ASCT). METHODS: Sixty-eight MM patients submitted to bortezomib-based induction therapy and ASCT from January 2007 to July 2012 were retrospectively studied. Monoclonal protein was detected by immunofixation electrophoresis (IFE). RESULTS: Of all 68 patients, 33 (48.5%) patients had APB. At the first emergence of an APB, two patients with light chain type achieved CR and before transplantation, and thirty-one patients were after transplantation with median time of 104 (ranged 33-404) days. The median duration of APB appearance was 105 (ranged 35-801) days. Patients who developed APB compared with those without APB, had a significantly higher CR plus very good partial response (VGPR) rates (100.0% vs 85.7%%, P=0.017) and CR rates (87.9% vs 62.9%) (P=0.03). There were no significant differences in gender, age, HGB, ALB, ß2-microglobulin, M protein type, Durie-Salmon and ISS stages, the case number of first line or second line treatment, induction courses of bortezomib-based regimen, and the mode of ASCT. With a median follow-up of 33.4 (ranged 7.0-71.7) months, patients with APB tended to have a longer overall survival (OS) versus non-APB patients, although no significant difference obtained (P＞0.05). Among APB patients, OS was longer in patients whose appearance of APB occurred ＜6 months after transplantation than those ≥ 6 months, but the significant difference was not obtained yet (P＞0.05). CONCLUSIONS: Patients who developed APB had a significantly better response to bortezomib-based induction regimen followed ASCT. APB emergence has a good prognostic significance.

Sequential treatment with bortezomib plus dexamethasone followed by autologous hematopoietic stem cell transplantation in patients with multiple myeloma.

BACKGROUND: Whether the sequential treatment with bortezomib plus dexamethasone (BD) followed by autologous hematopoietic stem cell transplantation (ASCT) could extend the overall survival period in multiple myeloma patients is still not clear. Few large case studies about this therapeutics in multiple myeloma were reported in China. Our purpose was to assess the efficacy and adverse effects of sequential treatment with BD chemotherapy and ASCT in patients with multiple myeloma. METHODS: Fifty-three patients with newly diagnosed or relapsed/refractory multiple myeloma received BD as induction therapy before ASCT. Stem-cell mobilization was undertaken with cyclophosphamide 3 - 5 g/m(2) plus granulocyte colony-stimulating factor 300 µg/d. Target yield was 2.0×10(6) CD34(+) cells/kg. Conditioning for ASCT consisted of melphalan 200 mg/m(2). Thalidomide and/or a-interferon was used as post-transplantation maintenance treatment. RESULTS: The BD chemotherapy before transplantation was effective in 86.7% of the 53 patients, including 22.6% with complete remission (CR), 39.6% with near complete remission (nCR), and 24.5% with partial remission (PR). The best effect was achieved after two treatment courses. Most bortezomib-related adverse effects were classes 1 - 2. All patients were successfully mobilized after BD for autologous peripheral blood stem cell transplantation. The ASCT was effective in 96.3% of patients, including 49.1% with CR, 32.1% with nCR, and 15.1% with PR. The CR rate was significantly increased (49.1% vs. 22.6%, P < 0.05) by sequential ASCT. Within 27 (range, 6 - 53) months of follow-up, the efficacy of ASCT was maintained in 29 patients and further enhanced by post-transplantation maintenance treatment in four patients. Eleven patients died after transplantation. Among the patients undergoing BD/ASCT treatment, overall survival (OS) was significantly better in newly diagnosed patients in comparison to relapsed/refractory patients (P = 0.046). CONCLUSIONS: BD chemotherapy can be used as an induction therapy prior to ASCT in patients with multiple myeloma. Its rate of effectiveness is high and it alleviates symptoms quickly without affecting peripheral blood stem cell collection. The majority of adverse effects are mild (tolerable). Sequential BD with ASCT is the preferred option for transplant patients. First-line ASCT could prolong survival of newly diagnosed patients rather than delayed ASCT.

[BAFF level in bone marrow and expression of BAFF receptor on B cells in multiple myeloma patients].

This study was purposed to investigate the B cell-activating factor belonging to the TNF family (BAFF) and a proliferation-inducing ligand (APRIL) levels in bone marrow, and the BAFF receptor expression level on B cells in multiple myeloma (MM) patients, in order to explore the characteristics of B cells in bone marrow of MM patients. MM patients were studied before treatment (newly diagnosed group, 19 patients) and after treatment with improvement (stable group, 17 patients), 10 non-hematologic patients were selected as control (control group). The BAFF receptors (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) on B cell (CD19(+)), naive B cell (CD19(+)IgD(+)) and memory B cell (CD19(+)CD27(+)) of bone marrow in all groups were detected by flow cytometry. The BAFF, APRIL level in bone marrow supernatant were tested with ELISA. The results showed that the BAFF-R expression level on CD19(+) cells in newly diagnosed group were higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group and stable group, but BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group was higher than that in control group; the BAFF-R expression level on CD19(+)CD27(+) cells in newly group was higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in stable group and control group. There was no significant difference among the TACI expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in newly diagnosed group, stable group and control group. The bone marrow supernatant BAFF level in newly diagnosed group was higher than that in stable group and control group, but there was no significant difference between stable group and control group. There was no significant difference among the bone marrow TACI levels in newly diagnosed group, stable group and control group. It is concluded that both the bone marrow BAFF level and the BAFF-R expression level on CD19(+) cell, CD19(+)IgD(+) cells and CD19(+)CD27(+) cells in MM patients increase, which may help to stimulate B cells, thereby may relate with to MM pathogenesis.

[The efficacy and safety of bortezomib-based induction regimen before autologous hematopoietic stem cell transplantation in patients with multiple myeloma].

OBJECTIVE: To investigate the efficacy and safety of bortezomib-based induction regimen followed by autologous hematopoietic stem cell transplantation (ASCT) in patients with multiple myeloma (MM). METHODS: A retrospective analysis was performed upon clinical data of 62 MM patients who received bortezomib-based induction regimen followed by ASCT from June 2006 to June 2011. All patients were followed up to September 30, 2011. RESULTS: Overall response rate [complete remission (CR) + near complete remission (nCR) + partial remission (PR)], ≥ nCR rate (CR/nCR) and CR rate of post-induction with bortezomib-based regimen were 88.7%, 66.1% and 24.2%, respectively. After ASCT, CR rate and CR/nCR rate were increased to 50.0% and 82.3%, respectively, with significant differences (P = 0.003 and P = 0.032). The median time of neutrophil and platelet engraftment was 12.0 (9 - 43) days and 13.5 (0 - 120) days, respectively. Significances were found in neutrophil and platelet engraftment between MM patients with and without prior exposure to alkylating agents. Furthermore, engraftment of neutrophil and platelet in patients receiving peripheral blood stem cell transplantation were faster than those receiving bone marrow transplantation. No unexpected side effects occurred. The median time of follow-up was 26.5 (7-61) months. The median overall survival (OS) was not reached and the median progression-free survival (PFS) was 30 months. There were significant differences in OS and PFS between patients obtaining CR/nCR and those with ≤ PR before ASCT. CONCLUSIONS: Bortezomib-based induction regimen can improve the efficacy of ASCT in MM patients. The side effects are tolerant. Higher response quality before ASCT can translate to high rates of OS and PFS following high-dose therapy and stem cell transplantation.

Differentiation induction enhances bortezomib efficacy and overcomes drug resistance in multiple myeloma.

AIM: It is of clinical importance to find methods to overcome bortezomib resistance. In the current study, we clarified the relationship between resistance to bortezomib and the differentiation status of myeloma cells, and explored the feasibility of induction of differentiation in overcoming bortezomib resistance in myeloma. METHODS: Cell morphology, immunoglobulin light-chain protein secretion levels, and XBP-1 expression were used to evaluate the differentiation status of myeloma cells. Low dose 2-ME2 alone or in combination with ATRA was used to induce differentiation in myeloma cells. RESULTS: The differentiation status of myeloma cells was related to myeloma sensitivity to bortezomib. After successful induction of differentiation, the myeloma cells were more sensitive to bortezomib with decreased growth and an increased rate of apoptosis. Induction of differentiation increased the proteasome workload in myeloma cells by increasing immunoglobulin secretion, while reducing proteasome capacity by decreasing proteasome activity. The imbalance between increased proteasome workload and decreased proteasome capacity is a possible mechanism by which induction of differentiation overcomes myeloma resistance to bortezomib. CONCLUSION: The current study demonstrated, for the first time, that myeloma differentiation status is associated with myeloma sensitivity to bortezomib and that induction of differentiation can overcome myeloma resistance to bortezomib.

[Maintenance therapies of thalidomine and interferon-α in multiple myeloma].

OBJECTIVE: To explore the efficacies and toxicities in multiple myeloma (MM) patients on the maintenance therapies of thalidomide and interferon-α so as to seek the optimal chemotherapeutic regimen. METHODS: A retrospective analysis was conducted for 57 MM patients on the maintenance therapies of thalidomide and interferon-α after introduction and consolidation. And 56 MM patients without maintenance therapy were enrolled as the control group. RESULTS: The values of progression-free survival (PFS) and overall survival (OS) were significantly longer in the maintenance group and this translated into an improved estimated 3-year PFS of 75.4% (71.8%, 83.3%) versus 23.2% in the control group (P < 0.01). The estimated 4-year OS was higher in the maintenance group [89.5% (89.7%, 88.9%) vs 33.9%, P < 0.01]. No statistically significant differences existed among different maintenance groups in terms of PFS and OS. The administration of maintenance therapy extended both PFS and OS for MM patients of various M-proteins (P < 0.05). However, in the thalidomide group, PFS and OS were extended only in MM of immunoglobulin G (IgG) and immunoglobulin A (IgA) but not in light-chain patients. Furthermore, the MM patients of Durie-Salmon (DS) stages II and III and international staging system (ISS) stages II and III extended PFS and OS through maintenance (P < 0.05). While in those of ISS stage I, the differences were insignificant in terms of PFS and OS between two groups. The results were similar between the thalidomide and control groups. The patients achieving a partial remission (PR) or higher response level benefited from the maintenance therapy in terms of PFS and OS (P < 0.05). In the thalidomide group, the patients with below PR prolonged OS (P = 0.031) but did not achieve a longer PFS (P = 0.091). Both PFS and OS were extended through maintenance therapy after either stem cell transplantation or consolidation chemotherapies (P < 0.05). There was no significant difference in terms of PFS and OS between MM patients without maintenance therapy after transplantation and those without transplantation. The adverse effects of thalidomide, milder than those of interferon-α, could be tolerated in most patients. The incidence and severity of adverse effects showed no significant difference between the combination maintenance and single agent therapies. CONCLUSION: The maintenance therapies of thalidomide and interferon-α could improve the profiles of PFS and OS in MM patients. And there was no significant difference between them in terms of PFS and OS. However, the maintenance therapy of thalidomide is a better option due to its convenient application, milder adverse effects, reasonable cost and better efficacies in MM patients not achieving PR or receiving induction therapy without bortezomib or without transplantation.

[Expression and significance of N-cadherin and ß-catenin protein in osteosarcoma].

OBJECTIVE: To evaluate the expression of N-cadherin and ß-catenin protein and their relationship with clinicopathological characteristics of osteosarcoma. METHODS: The expressions of N-cadherin and ß-catenin at protein level were detected by immunohistochemical staining in 54 cases of osteosarcoma, 11 cases of osteoid osteoma, 7 cases of osteoblastoma and 8 cases of newly formed bone in nonmalignant bone diseases. The relationship between the two indexes and clinicopathological characteristics of osteosarcoma was analyzed. RESULTS: In newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the positive expression rate of N-cadherin protein was 75.0%, 71.4%, 63.6% and 35.2%, respectively. The positive expression rate of N-cadherin protein in osteosarcoma was significantly lower than that in osteoid osteoma, osteoblastoma and newly formed bone in nonmalignant bone diseases (P = 0.035). The positive expression rate of N-cadherin protein in osteosarcoma cases with pulmonary metastasis was lower than that in cases without (21.7% vs. 56.3%, P = 0.027). The positive expression rate of N-cadherin protein in osteosarcoma cases died in two years was lower than that in cases lived for more than two years (18.2% vs. 50.0%, P = 0.024). In newly formed bone, osteoblastoma, osteoid osteoma and osteosarcoma, the aberrant expression rate of ß-catenin protein was 12.5%, 28.6%, 27.3% and 66.7%, respectively. The aberrant expression rate of ß-catenin protein in osteosarcoma was significantly higher than that in osteoid osteoma, osteoblastoma and newly formed bone (P = 0.002). Aberrant expression rate of ß-catenin in osteosarcoma cases with pulmonary metastasis was higher than that without (82.6% vs. 43.8%, P = 0.011). An inverse correlation was found between the aberrant expression of ß-catenin and N-cadherin expression in osteosarcoma(r = -0.302, P = 0.027). CONCLUSION: The positive expression rate of N-cadherin is decreased in osteosarcoma while aberrant expression rate of ß-catenin increased. The expression of N-cadherin protein is closely correlated with the metastasis and prognosis of osteosarcoma, but the expression of ß-catenin protein is merely correlated with the metastasis of osteosarcoma.

[Clinical observation of therapeutic efficacy and study of conjunctival impression cytology after pterygium surgery].

OBJECTIVE: To compare the pathologic changes, the recurrence rate and the ocular surface damage after 4 different types of pterygium surgical procedures. METHODS: It is a prospective study. From Mar. 2006 to Mar. 2008, 84 patients (93 eyes) from Shi Jia-zhuang Center Hospital were selected and separated into 4 groups as follows, simple excision group, 24 patients (27 eyes); excision with conjunctival allograft group, 21 patients (22 eyes) excision with autologous limbus stem cell transplantation group, 18 patients (20 eyes) and excision with mitomycin (MMC) group, 21 patients (24 eyes). Slit lamp microscope examination and impression cytology were performed 1 day before the surgery and repeated 30 and 90 days after the surgery. Multivariate analysis of variance was used to analyze these data with statistical software SPSS 11.0. P value less than 0.05 was considered statistically significant. RESULTS: Before the surgery, slight conjunctival epithelium keratinization, mild acantholysis and decrease in goblet cells density were identified in all patients (248.8 +/- 97.3, 256.2 +/- 75.1, 236.1 +/- 87.9, 245.1 +/- 81.0; F = 0.029, P = 0.993). All of these changes decreased 1 month after the surgery. MMC group showed more goblet cells than the other 3 groups, but the difference was not statistically significant (F = 0.747, P = 0.554). Three months after the surgery, all of these changes in MMC group (112.1 +/- 56.8) were significantly more severe than those in other 3 groups (309.6 +/- 77.0, 314.1 +/- 68.9, 317.4 +/- 73.2; F = 6.337, P = 0.017) and no difference could be detected between these 3 groups. There were 5 recurrences in simple excision group, 3 recurrences in MMC group, 1 recurrence in stem cell transplantation group and none in conjunctival allograft group. CONCLUSIONS: There are fewer recurrent cases in conjunctival allograft group and stem cell transplantation group than the others. Severe ocular surface damage is present in MMC group.

[In vitro study of human multiple myeloma cell transfected with soluble vascular endothelial growth factor receptor-1 gene].

BACKGROUND AND OBJECTIVE: Vascular endothelial growth factor (VEGF) is the most important angiogenic factor of multiple myeloma (MM). This study was to investigate the effect of transfection of human soluble vascular endothelial growth factor receptor-1 (sFlt-1) gene on the proliferation of human MM cell line RPMI8226. METHODS: The recombinant plasmid pcDNA3-sFlt-1 was constructed and transfected into RPMI8226 cells. The expression of sFlt-1 was identified by reverse transcription-polymerase chain reaction (RT-PCR) and ELISA. The effects of sFlt-1 protein on the proliferation and VEGF expression of RPMI8226 cells were investigated by MTT assay and ELISA, respectively. RESULTS: The recombinant plasmid pcDNA3-sFlt-1 was successfully transfected into RPMI8226 cells. sFlt-1 protein expression was identified by ELISA, which inhibited the proliferation of RPMI8226 cells and reduced VEGF concentration in the culture supernatant. CONCLUSION: RPMI8226 cells can express sFlt-1 protein with high biological activity when transfected with the sFlt-1 gene, which inhibits the proliferation of RPMI8226 cells.

The regulatory effect of the p38 signaling pathway on valdecoxib-induced apoptosis of the Eca109 cell line.

Valdecoxib is a second generation selective COX-2 inhibitor that can induce cell apoptosis in a variety of cell types, but its precise regulatory mechanism is unknown. Apoptosis of Eca109 cells and p38 mRNA expression were investigted. The expression of p-p38MAPK, Fas and FasL proteins were detected by immunohistochemical staining and FCM. Valdecoxib increased the apoptosis rate of Eca109 cells. Fas and FasL protein expression was up-regulated in the valdecoxib groups, while SB203580 partly inhibited the valdecoxib-induced overexpression. Valdecoxib increased p38MAPK expression, while SB203580 inhibited the overexpression of this protein and the apoptosis rate decreased. The expression of Fas, FasL and p38MAPK protein were positively correlated with the apoptotic rate. In conclusion, valdecoxib activates the p38MAPK pathway, thus up-regulating expression of the Fas and FasL proteins, which may be one of the mechanisms through which valdecoxib induces apoptosis.

[The clinical features and risk factors for invasive fungal infection in multiple myeloma.].

OBJECTIVE: To study the clinical features and risk factors of invasive fungal infection (IFI) in multiple myeloma (MM). METHODS: Three hundred and fifty-seven cases of MM were retrospectively analyzed for IFI, clinical features, complicating diseases, treatment of fungus and side effect of anti-fungal drugs. RESULTS: Forty-four cases (12.3%) of IFI were diagnosed. Three of them were diagnosed definitely, 8 clinically and 33 probably. Ten cases incurred IFI in the induction therapy, 4 in platform, 27 in progress and 3 in the treatment with autologous stem cell transplantation. The lung was the commonest site of infection (50.0%). The total effective rates of amphotericin B liposome, voriconazole, itraconazole, caspofungin and fluconazol were 83.3%, 75.0%, 78.9%, 75.0% and 57.1% respectively (P = 0.493). In a multivariate analysis, independent factors significantly associated with IFI were diabetes (P = 0.035, OR 2.527, 95%CI 1.005 - 6.052), dialysis (P = 0.022, OR 2.768, 95%CI 1.161 - 6.600), persistent agranulocytosis (P = 0.019, OR 3.215, 95%CI 1.200 - 7.407), broad-spectrum antibiotic therapy (P = 0.009, OR 3.350, 95%CI 1.353 - 8.295) and fludarabine treatment (P = 0.001, OR 4.669, 95%CI 1.813 - 12.023). CONCLUSIONS: Patients with MM are in high risk of IFI. The lung is the commonest site of infection. The therapeutic effect was similar with itraconazole, voriconazole, caspofungin and amphotericin B liposome in MM patients with complicating IFI. The risk factors for IFI in MM were diabetes, dialysis, persistent agranulocytosis and the use of broad-spectrum antibiotics and fludarabine.

[Expression and clinical significance of COX-2, p-Stat3, and p-Stat5 in esophageal carcinoma].

BACKGROUND & OBJECTIVE: Cyclooxygenase-2 (COX-2) and signal transducers and activators of transcription (STAT) are closely correlated to the genesis of tumors. This study was to investigate the expression and clinical significance of COX-2, p-Stat3 and p-Stat5 (the activated forms of Stat3 and Stat5) in various lesions of esophageal tissues, and to analyze their correlations to clinicopathologic features of esophageal squamous cell carcinoma (ESCC). METHODS: The expression of COX-2, p-Stat3, and p-Stat5 in 59 specimens of ESCC, 24 specimens of squamous dysplasia, and 18 specimens of normal squamous epithelium was examined by SP immunohistochemistry. Their correlations to clinicopathologic features of ESCC were analyzed. RESULTS: The protein level of COX-2 was significantly higher in ESCC and squamous dysplasia than in normal squamous epithelium (2.10+/-1.77 and 1.85+/-1.24 vs. 0.83+/-0.46, P<0.05). The protein level of p-Stat3 was 0 in normal squamous epithelium, 0.76+/-0.59 in squamous dysplasia, and 2.83+/-1.27 in ESCC. The protein level of p-Stat5 was 1.98+/-0.78 in normal squamous epithelium, 3.92+/-0.41 in squamous dysplasia, and 5.02+/-0.34 in ESCC. There were significant differences among the 3 groups (P<0.05). In ESCC, COX-2 expression was correlated to lymph node metastasis and differentiation (P<0.05); p-Stat3 expression was correlated to tumor invasion depth (P<0.05); but p-Stat5 expression had no correlation to clinicopathologic features. COX-2 expression was positively correlated to both p-Stat3 expression and p-Stat5 expression in ESCC. CONCLUSIONS: The up-regulation of COX-2, p-Stat3, and p-Stat5 may be correlated to the carcinogenesis of ESCC. The activation of Stat3 is correlated to the aggressive behavior of ESCC.

[Expression and clinical significance of vascular endothelial growth factor and its receptors in multiple myeloma].

BACKGROUND & OBJECTIVE: Bone marrow angiogenesis plays an important role in the development and prognosis of multiple myeloma (MM). Vascular endothelial growth factor (VEGF) is the most potent stimulatory factor in angiogenesis. This study was to examine the expression of VEGF and its receptors in MM, and analyze their correlations to the tumorigenesis and development of MM. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mRNA expression of VEGF, Flt-1, and KDR in bone marrow samples from 12 newly diagnosed MM patients, 23 relapsed/refractory MM patients, 16 control people, and in KM3 cells. RESULTS: The positive rates of VEGF and Flt-1 were significantly higher in MM patients than in control people (62.9% vs. 18.8%, P<0.01; 80% vs. 31.3%, P<0.01). The mRNA levels of VEGF and Flt-1 were significantly higher in MM patients than in control people (0.41+/-0.19 vs. 0.06+/-0.01, P<0.05; 0.60+/-0.33 vs. 0.08+/-0.03, P<0.01). The positive rates of VEGF and Flt-1 mRNA were not significantly different between newly diagnosed and relapsed/refractory MM patients (66.7% vs. 60.9%, P>0.05; 83.3% vs. 78.3%, P>0.05), and between stage II MM and stage III MM (50% vs. 73.7%, P>0.05; 81.3% vs. 78.9%, P>0.05). The mRNA levels of VEGF and Flt-1 were significantly higher in relapsed/refractory MM patients than in newly diagnosed MM patients (0.49+/-0.20 vs. 0.28+/-0.04, P<0.05; 0.70+/-0.38 vs. 0.41+/-0.06, P<0.05), and were significantly higher in stage III MM than that in stage II MM (0.48+/-0.19 vs. 0.28+/-0.09, P<0.05; 0.75+/-0.35 vs. 0.41+/-0.21, P<0.05). KDR was detected only in 3 MM patients and was not detected in control group. CONCLUSION: High expression of VEGF and Flt-1 mRNA is associated with the development of MM.