[Diagnosis and treatment status of perioperative anemia in patients with gastrointestinal neoplasms: a multi-center study in Hubei Province].

Objective: To investigate the incidence and treatment of perioperative anemia in patients with gastrointestinal neoplasms in Hubei Province. Methods: The clinicopathological data of 7 474 patients with gastrointestinal neoplasms in 62 hospitals in 15 cities (state) of Hubei Province in 2019 were collected in the form of network database. There were 4 749 males and 2 725 females. The median age of the patients was 62 years (range: 17 to 96 years). The hemoglobin value of the first time in hospital and the first day after operation was used as the criterion of preoperative anemia and postoperative anemia. Anemia was defined as male hemoglobin <120 g/L and female hemoglobin <110.0 g/L, mild anemia as 90 to normal, moderate anemia as 60 to <90 g/L, severe anemia as <60 g/L. The t test and χ2 test were used for inter-group comparison. Results: The overall incidence of preoperative anemia was 38.60%(2 885/7 474), and the incidences of mild anemia, moderate anemia and severe anemia were 25.09%(1 875/7 474), 11.37%(850/7 474) and 2.14%(160/7 474), respectively. The overall incidence of postoperative anemia was 61.40%(4 589/7 474). The incidence of mild anemia, moderate anemia and severe anemia were 48.73%(3 642/7 474), 12.20%(912/7 474) and 0.47%(35/7 474), respectively. The proportion of preoperative anemia patients receiving treatment was 26.86% (775/2 885), and the proportion of postoperative anemia patients receiving treatment was 14.93% (685/4 589). The proportions of preoperative anemia patients in grade ⅢA, grade ⅢB, and grade ⅡA hospitals receiving treatment were 26.12% (649/2 485), 32.32% (85/263), and 29.93% (41/137), and the proportions of postoperative anemia patients receiving treatment were 14.61% (592/4 052), 22.05% (73/331), and 9.71% (20/206). The proportion of intraoperative blood transfusion (16.74% (483/2 885) vs. 3.05% (140/4 589), χ²=434.555, P<0.01) and the incidence of postoperative complications (17.78% (513/2 885) vs. 14.08% (646/4 589), χ²=18.553, P<0.01) in the preoperative anemia group were higher than those in the non-anemia group, and the postoperative hospital stay in the preoperative anemia group was longer than that in the non-anemia group ((14.1±7.3) days vs. (13.3±6.2) days, t=5.202, P<0.01). Conclusions: The incidence of perioperative anemia in patients with gastrointestinal neoplasms is high. Preoperative anemia can increase the demand for intraoperative blood transfusion and affect the short-term prognosis of patients. At present, the concept of standardized treatment of perioperative anemia among gastrointestinal surgeons in Hubei Province needs to be improved.

[Curative effects and influenced factors of primary chemotherapy with single-agent methotrexate on low-risk gestational trophoblastic neoplasia].

Objective: To evaluate the efficacy of primary chemotherapy with single-agent methotrexate (MTX) for low-risk gestational trophoblastic neoplasia and to analysis the influenced factors. Methods: We retrospectively reviewed 259 cases with low-risk gestational trophoblastic neoplasia whose primary chemotherapies were MTX 0.4 mg·kg(-1) (maximum 25 mg) daily for 5 days every other week. Patients' data between January 2001 and June 2015 was collected and the relationships of different factors to outcomes of chemotherapy were also evaluated. Results: 183 of the 259 patients (70.66%, 183/259) achieved complete primary remission and all patients achieved complete remission after salvage chemotherapy. Univariate analysis showed that FIGO score, serum level of HCG before treatment and interval months from previous pregnancy were significantly associated with outcome of chemotherapy (P=0.001, 0.018, 0.014 respectively). Logistic regression analysis showed that the FIGO score (OR=4.094) and antecedent pregnancy (OR=0.268) were two independent factors predictive for the outcome of chemotherapy. Conclusions: Primary chemotherapy with single-agent MTX may still be one of the options for patients with low risk GTN. The FIGO score and antecedent pregnancy are two independent risk factors of outcome of single-agent MTX chemotherapy.

Resistance training vs. aerobic training and role of other factors on the exercise effects on visceral fat.

In the last 25 years, obesity has reached epidemic levels of prevalence. It has even affected the children such that rates of severe childhood obesity that have almost tripled in numbers. These numbers are alarming because of the known fact that obesity is associated with an increased risk of several comorbidities as well as with an increased risk of premature death. Almost since the beginning, exercise has been known to play a key role in the prevention and treatment of overweight and the non-pharmacological treatment of dyslipidemia. However, the effects of exercise on obesity seems to be dynamic and influenced by several other factors. These factors can be related to exercise or to the associated comorbidities. In this review we will address following factors: (1) The type of exercise which could be either aerobic or resistance training (2) The volume or amount of training (3) Intensity of training and (4) The effect of comorbidity of diabetes mellitus. We will observe that all of these factors modify the effect of exercise on the visceral fat.

HER-2-induced PI3K signaling pathway was involved in the pathogenesis of gastric cancer.

Human epidermal growth factor receptor-2 (HER-2) overexpression was closely associated with the tumor growth and invasion, we here aimed to investigate the mechanism of HER-2 mediation in the pathogenesis of gastric cancer (GC). We first detected the expression of HER-2 in GC cell line SGC-7901 and then examined the levels of nuclear factor-κB (NF-κB), matrix metalloproteinase-9 (MMP-9) and intercellular adhesion molecule-1 (ICAM-1) and the association between them by molecular methods. Statistical analysis was used to compare the significance. We further detected the possible molecular mechanism involved in their relationship in the SGC-7901 genesis. The MMP-9, NF-κB and secretory type (s-ICAM-1) levels were significantly greater in peripheral blood serum from SGC-7901 than healthy control GES-1 (P<0.01). ICAM-1, MMP-9 and NF-κB mRNA and protein levels were more highly expressed in SGC-7901 than healthy control GES-1. The expression levels of NF-κB, MMP-9 and ICAM-1 were positively related in GC cell line SGC-7901, which was HER-2 positive. The HER-2 positive SGC-7901 secreted more transforming growth factor beta 1 (TGF-ß1) and resultantly activated MMP-9 to enhance s-ICAM-1 secretion and further studies showed that phosphatidylinositol-3 kinase (PI3K)/Akt/NF-κB signaling pathway was involved in GC pathogenesis. The GC cells that express the HER-2 oncogene spur the activation of NF-κB that can upregulate the expression of ICAM-1 and induce the expression of MMP-9, which hydrolyzes ICAM-1 into s-ICAM-1 to promote tumor immune escape. TGF-ß1-induced PI3K/Akt/NF-κB signaling pathway was involved in the pathogenesis of GC and they could be a new target for cancer therapy. The GC cells that express the HER-2 oncogene spur the activation of NF-κB that can upregulate the expression of ICAM-1 and induce the expression of MMP-9, which hydrolyzes ICAM-1 into s-ICAM-1 to promote tumor immune escape. TGF-ß1-induced PI3K/Akt/NF-κB signaling pathway was involved in the pathogenesis of GC and they could be a new target for cancer therapy.

Effect of the route of foot-and-mouth disease virus infection of piglets on the course of disease.

Three different routes of Foot-and-mouth disease virus (FMDV) infection of piglets, namely intranasal (i.n.) through drops, intradermal (i.d.) into the foot, and intramuscular (i.m.) were compared regarding the onset and severity of the disease. The results showed that the i.d. injection of the virus resulted in the fastest onset of the disease. The i.m. injection led to a delayed onset, but the final effect was identical with i.d. injection. Moreover, the i.m. injection was simpler to perform and easier to evaluate. Therefore, the i.m. injection of piglets is recommended as the optimal infection route for evaluation of the FMDV vaccine potency.

EMA-CO chemotherapy for high-risk gestational trophoblastic neoplasia: a clinical analysis of 54 patients.

This study was designed to analyze the outcomes of chemotherapy for high-risk gestational trophoblastic neoplasia (GTN) with EMA-CO regimen as primary and secondary protocol in China. Fifty-four patients with high-risk GTN received 292 EMA/CO treatment cycles between 1996 and 2005. Forty-five patients were primarily treated with EMA-CO, and nine were secondarily treated after failure to other combination chemotherapy. Adjuvant surgery and radiotherapy were used in the selected patients. Response, survival and related risk factors, as well as chemotherapy complications, were retrospectively analyzed. Thirty-five of forty-five patients (77.8%) receiving EMA-CO as first-line treatment achieved complete remission, and 77.8% (7/9) as secondary treatment. The overall survival rate was 87.0% in all high-risk GTN patients, with 93.3% (42/45) as primary therapy and 55.6% (5/9) as secondary therapy. The survival rates were significantly different between two groups (chi(2)= 6.434, P =0.011). Univariate analysis showed that the metastatic site and the number of metastatic organs were significant risk factors, but binomial distribution logistic regression analysis revealed that only the number of metastatic organs was an independent risk factor for the survival rate. No life-threatening toxicity and secondary malignancy were found. EMA-EP regimen was used for ten patients who were resistant to EMA-CO and three who relapsed after EMA-CO. Of those, 11 patients (84.6%) achieved complete remission. We conclude that EMA-CO regimen is an effective and safe primary therapy for high-risk GTN, but not an appropriate second-line protocol. The number of metastatic organs is an independent prognostic factor for the patient with high-risk GTN. EMA-EP regimen is a highly effective salvage therapy for those failing to EMA-CO.

Investigation of the immunosuppressive activity of artemether on T-cell activation and proliferation.

BACKGROUND AND PURPOSE: Artemisinin and its derivatives exhibit potent immunosuppressive activity. The purpose of the current study was to examine the immunosuppressive activity of artemether directly on T lymphocytes and to explore its potential mode of action. EXPERIMENTAL APPROACH: In vitro, T-cell proliferation was measured using [(3)H]-thymidine incorporation assay in cells stimulated with ConA, alloantigen and anti-CD3 antibody. CFSE-labeled cell division and cell cycle distribution were monitored by flow cytometry. In vivo, the effects of artemether were evaluated in delayed-type hypersensitivity (DTH) and purified T-cell responses to ovalbumin in ovalbumin-immunized mice. The activation of extracellular signal-regulated kinase1/2 (ERK1/2) and Raf1 were assessed by Western blot analysis and the activation of Ras was tested in pull-down assays. KEY RESULTS: We show that, in vitro, artemether suppressed ConA- or alloantigen-induced splenocyte proliferation, influenced production of the cytokines IL-2 and IFN-gamma and inhibited cell cycle progression through the G0/G1 transition. In vivo, administration of artemether attenuated CD4 T-cell-mediated DTH reaction, and suppressed antigen-specific T-cell response in immunized mice. Further experiments showed that, treatment with artemether impaired both antigen- and anti-CD3-induced phosphorylation of ERK. In primary T cells, artemether profoundly inhibited anti-CD3-induced phosphorylation of Raf1 and activation of Ras. CONCLUSIONS AND IMPLICATIONS: This study provided experimental evidence of the immunosuppressive effects of artemether directly on T cells both in vitro and in vivo. Its immunosuppressive mechanism involved inhibition of the activation of the Ras-Raf1-ERK1/2 protein kinase cascade in T cells.

Mycophenolate mofetil therapy for children with lupus nephritis refractory to both intravenous cyclosphosphamide and cyclosporine.

We describe mycophenolate mofetil (MMF), a new immunosuppressive agent, to be a therapy of two children with lupus nephritis which were refractory to both cyclophosphamide (CyP) and cyclosporine (CsA). After 11- to 12-month course of MMF treatment, all clinical symptoms of lupus disappeared and serum antibodies became negative. MMF might be a promising curative for cyclophosphamide-resistant lupus nephritis in children. Cyclophosphamide intravenous bolus therapy is generally considered to be the treatment for patients with lupus nephritis. However, there is little guidance about what to do if such therapy fails. Recently, a new immunosuppressive agent, mycophenolate mofetil (MMF), has been used to treat cyclophosphamide-resistant lupus nephritis [Dooley et al. 1999, Gaubitz et al. 1999, Glicklich and Acharga 1998] in adults and has been recognized as a promising curative for lupus nephritis. Up to now, MMF has been adopted widely with solid organ transplantation to prevent or reverse acute rejection [Mathew 1998, Morris-Stiff and Jurewicz 1998] and has been used successfully to treat for rheumatoid arthritis refractory to a variety of other drugs. But there is no report about MMF treatment in children with cyclophosphamide-resistant lupus nephritis. We describe our experience with MMF treatment in two Chinese children with lupus nephritis that were refractory not only to cyclophosphamide but also to cyclosporine.

Flow cytometric assessment of the cellular pharmacokinetics of fluorescent drugs.

Development of multidrug resistance (MDR) in cancer cells decrease net doxorubicin uptake as a result of either increased efflux, or decreased intracellular sequestration, or decreased membrane permeability. Kinetic parameters of drug uptake can distinguish among these forms of altered transport. Cellular uptake of fluorescent drugs was monitored by a flow cytometric assay using a rapid-injection system and analyzed with a three-compartment model in which rapid diffusion from extracellular fluid into the cell was followed by uptake into a nonexchangeable pool. In agreement with our recent studies of 14C-doxorubicin distribution (Dordal et al.: J Pharmacol Exp Ther 271:1286-1290, 1994), sequestration of doxorubicin was decreased 2.7-fold in P-glycoprotein-expressing SU-4R lymphoma cells compared to drug-sensitive SU-4 cells (14.0 +/- 4.8 vs. 5.0 +/- 0.9 nl s-1) without a change in membrane permeability or evidence of active efflux. In contrast, sequestration of the highly fluorescent dye rhodamine 123 was decreased 20-fold (17.1 +/- 8.3 vs. 0.9 +/- 0.8 nl s-1). Resistant cells were significantly less permeable to rhodamine than sensitive cells (3.8 +/- 1.2 vs. 10.2 +/- 2.6 x 10(5) cm2 s-1), and rhodamine efflux was increased by 24%. Thus, SU-4R cells exhibit multiple alterations that cause decreased intracellular drug concentrations, of which decreased sequestration is quantitatively the most significant.

Erythrocyte membrane lipid composition fluidity in patients with essential hypertension.

The lipid composition and fluidity of erythrocyte membrane in 36 patients with essential hypertension were examined. The results showed that either cholesterol/phospholipid molar ratio or lipid peroxides content was significantly increased (P < 0.01 or 0.001), and the superoxide dismutase activity, contents of four classes of phospholipids, i.e., phosphatidyl ethanolamine, phosphatidylcholine, phosphatidylserine and sphingomyelin of erythrocyte membrane, and lipid fluidity were significantly decreased (P < 0.05, < 0.01 or < 0.001) as compared with those in 35 normotensive control subjects. These results suggested that the changes of lipid composition and fluidity might associate with the decreased activities of cation transport systems in cell membranes and play an important role in the pathogenesis of essential hypertension.

Effects of gossypol on the activity of kidney (Na+ + K+)-ATPase and the functions of erythrocyte membrane.

Gossypol extracted from cottonseed oil, reputed to exert contraceptive action in males, may induce a side effect of hypokalemia. The cellular and molecular mechanisms by which gossypol produces hypokalemia are not quite understood. We have examined the inhibitory effect of gossypol on the activity of (Na+ + K+)-ATPase purified from the outer medulla of rabbit kidney, the half maximal inhibitory concentration (IC50) is 6.5 micron. The kinetic studies using this enzyme preparation show that gossypol is noncompetitive with ATP, Mg2+, Na+ and K+ with apparent Ki of 15.0, 13.0, 14.0 and 4.4 micron, respectively. On the other hand, in order to estimate the effects of gossypol on membrane transport of intact cells, we have investigated the effects of gossypol on the activity of (Na+ + K+)-ATPase, membrane integrity and permeability of human erythrocytes. It shows that gossypol inhibited the enzyme activity (greater than 5 micron) and expressed the hemolysis (greater than 50 micron) in vitro in a concentration-dependent manner, and increased the K+-efflux of the cells (10-40 micron). The above effects are antagonized by 1-2% bovine serum albumin. The data demonstrate that gossypol is a specific and potent membrane active agent. These results may be relevant to the in vivo actions of gossypol.

PIP: Hypokalemia has been demonstrated to be the major side effect of gossypol administration for contraceptive purposes; however, the cellular and molecular mechanisms involved in this effect are unknown. This study investigated the effect of gossypol on the activity of (Na+ and K+)-ATPase purified from the rabbit kidney and the functions of erythrocyte membrane. Kinetic findings indicated gossypol is noncompetitive with ATP, Mg2+, Na+, and K+. On the other hand, gossypol inhibited the enzyme activity of (Na+ and K+)=ATPase, expressed the hemolysis in vitro in a concentration-dependent manner, and increased the K+ efflux of the cells. These effects were antagonized by 1-2% serum bovine albumin. These findings demonstrate that gossypol is a specific and potent membrane active agent capable of injuring the cell membrane.

The locus of nucleotide specificity in the reaction mechanism of (Na+ + K+)-ATPase determined with ATP and GTP as substrates.

ATP and GTP have been compared as substrates for (Na+ + K+)-ATPase in Na+-activated hydrolysis, Na+-activated phosphorylation, and the E2K----E1K transition. Without added K+ the optimal Na+-activated hydrolysis rates in imidazole-HCl (pH 7.2) are equal, but are reached at different Na+ concentrations: 80 mM Na+ for GTP, 300 mM Na+ for ATP. The affinities of the substrates for the enzyme are widely different: Km for ATP 0.6 microM, for GTP 147 microM. The Mg-complexed nucleotides antagonize activation as well as inhibition by Na+, depending on the affinity and concentration of the substrate. The optimal 3-s phosphorylation levels in imidazole-HCl (pH 7.0) are equally high for the two substrates (3.6 nmol/mg protein). The Km value for ATP is 0.1-0.2 microM and for GTP it ranges from 50 to 170 microM, depending on the Na+ concentration. The affinity of Na+ for the enzyme in phosphorylation is lower with the lower affinity substrate: Km (Na+) is 1.1 mM with ATP and 3.6 mM with GTP. The GTP-phosphorylated intermediate exists, like the ATP-phosphorylated intermediate, in the E2P conformation. Addition of K+ increases the optimal hydrolytic activity 30-fold for ATP (at 100 mM Na+ + 10 mM K+) and 2-fold for GTP (at 100 mM Na+ + 0.16 mM K+). K+ greatly increases the Km values for both substrates (to 430 microM for ATP and 320 microM for GTP). Above 0.16 mM K+ inhibits GTP hydrolysis. GTP does not reverse the quenching effect of K+ on the fluorescence of the 5-iodoacetamidofluorescein-labeled enzyme. ATP fully reverses this effect, which represents the transition from E1K to E2K. Hence GTP is unable to drive the E2K----E1K transition.

Thiophosphorylation of (Na + K+)-ATPase yields an ADP-sensitive phosphointermediate.

1) Treatment of (Na+ + K+)-ATPase from rabbit kidney outer medulla with the gamma-35S labeled thio-analogue of ATP in the presence of Na+ + Mg2+ and the absence of K+ leads to thiophosphorylation of the enzyme. The Km value for [gamma-S]ATP is 2.2 microM and for Na+ 4.2 mM at 22 degrees C. Thiophosphorylation is a sigmoidal function of the Na+ concentration, yielding a Hill coefficient nH = 2.6. (2) The thio-analogue (Km = 35 microM) can also support overall (Na+ + K+)-ATPase activity, but Vmax at 37 degrees C is only 1.13 mumol X (mg protein)-1 X h-1 or 0.09% of the specific activity for ATP (Km = 0.43 mM). (3) The thiophosphoenzyme intermediate, like the natural phosphoenzyme, is sensitive to hydroxylamine, indicating that it also is an acylphosphate. However, the thiophosphoenzyme, unlike the phosphoenzyme, is acid labile at temperatures as low as 0 degree C. The acid-denatured thiophosphoenzyme has optimal stability at pH 5-6. (4) The thiophosphorylation capacity of the enzyme is equal to its phosphorylation capacity, indicating the same number of sites. Phosphorylation by ATP excludes thiophosphorylation, suggesting that the two substrates compete for the same phosphorylation site. (5) The (apparent) rate constants of thiophosphorylation (0.4 s-1 vs. 180 s-1), spontaneous dethiophosphorylation (0.04 s-1 vs. 0.5 s-1) and K+-stimulated dethiophosphorylation (0.54 s-1 vs. 230 s-1) are much lower than those for the corresponding reactions based on ATP. (6) In contrast to the phosphoenzyme, the thiophosphoenzyme is ADP-sensitive (with an apparent rate constant in ADP-induced dethiophosphorylation of 0.35 s-1, Km ADP = 48 microM at 0.1 mM ATP) and is relatively K+-insensitive. The Km for K+ in dethiophosphorylation is 0.9 mM and in dephosphorylation 0.09 mM. The thiophosphoenzyme appears to be for 75-90% in the ADP-sensitive E1-conformation.