Efficacy of Trastuzumab + Cisplatin Combined with Irinotecan on the Quality of Life of Patients with Advanced Her-2 Positive Gastric Cancer.

Objective: To observe the effect of trastuzumab and cisplatin combined with irinotecan in the treatment of advanced Her-2 positive gastric cancer and its influence on disease control rate. Methods: From January 2018 to January 2021, 120 patients with advanced Her-2 positive gastric cancer admitted to our hospital were selected as the research subjects. According to the treatment plan of the patients, they were divided into a control group and a joint group, with 60 cases in each group; the control group was given trastuzumab + cisplatin, the joint group was treated with irinotecan on this basis, and the clinical effects and disease control rate of the two groups were observed. Results: After treatment, there were 4 patients with CR in the joint group and 0 patients with CR in the control group. The ORR and DCR of the joint group were significantly higher than those of the control group (P < 0.05). The expression levels of CA199, CEA, and CA724 after treatment in the two groups were significantly reduced (P < 0.05), and the reduction in the joint group after treatment was more evident as compared with the control group (P < 0.05). The joint group witnessed better memory function, physical function, behavioral function, emotional function, and communication function than the control group (P < 0.05), and the scores of all dimensions of the two groups of patients after treatment were superior to those before treatment (P < 0.05). The occurrence of side effects was not statistically different between the two groups of patients (P > 0.05). The 1-year survival rate of the control group was 41.67%, the PFS was 6.33 ± 1.02 months, and the OS was 15.51 ± 2.16 months; the 1-year survival rate of the joint group was 43.33%, and the PFS was 8.05 ± 1.07 months, and OS was 16.03 ± 2.44 months; there was no significant difference in the 1-year survival rate between the two groups (P > 0.05), the difference in PFS between the groups was significant (t = 9.013, P < 0.001), and the difference in OS between the groups was not significant (t = 1.236, P=0.219). Conclusion: Trastuzumab + cisplatin combined with irinotecan yields a promising result in the treatment of advanced gastric cancer. It can effectively regulate the expression level of tumor markers, delay disease progression, and improve the quality of life of patients. Moreover, irinotecan will not bring more toxic side effects.

Analysis of B-ultrasound and contrast-enhanced ultrasound characteristics of different hepatic neuroendocrine neoplasm.

BACKGROUND: Hepatic neuroendocrine neoplasm (hNEN) is a highly heterogeneous tumor. The exact identification of the source and malignant degree of hNEN is important. However, there is a lack of information regarding diagnosis of hNEN with imaging. In addition, no studies have compared the imaging between hNEN and hepatocellular carcinoma (HCC) and among different sources and malignant degrees of hNEN. AIM: To compare the ultrasound characteristics between hNEN and HCC and among different sources and malignant degrees of hNEN. METHODS: A total of 55 patients with hNEN were recruited and defined as the hNEN group. Among them, 35 cases of hNET were defined as the hNET group. Twenty cases of hepatic neuroendocrine carcinoma (hNEC) were defined as the hNEC group. Among the 55 lesions, 29 were transferred from the pancreas, 20 were from the gastrointestinal tract, and six were from other sites. In total, 55 patients with HCC were recruited and defined as the HCC group. The characteristic differences of B-mode ultrasound and contrast-enhanced ultrasound (CEUS) between hNEN and HCC and among different sources and malignant degrees of hNEN were compared. RESULTS: In the hNEN group, the proportions of multiple liver lesions, unclear borders, and high echo lesions were higher than those in the HCC group. The proportions of non-uniform echo and peripheral acoustic halo were lower than those in the HCC group (P < 0.05). The washout to iso-enhancement time and washout to hypo-enhancement time were lower than those in the HCC group (P < 0.05). The characteristics of B-ultrasound and CEUS among different sources of hNEN were similar, and the differences were not statistically significant (P > 0.05). B-mode ultrasound characteristics of hNET and hNEC were similar. The proportions of low enhancement at portal venous phase, non-uniform enhancement forms, and combined tumor vasculature in the hNEC group were larger than those in the hNEN group (P < 0.05). CONCLUSION: Compared with HCC, hNEN showed multiple intrahepatic lesions, uniform high echo, uniform high enhancement at arterial phase, and rapid washout. Low enhancement at portal venous phase, overall non-uniform enhancement form, and the proportion of combined tumor vasculature in hNEC were larger than those in hNET.

[Measures for Maintaining the Vitality of Peripheral Blood Hematopoietic Stem Cells in vitro].

OBJECTIVE: To explore the maintaining measures for the vitality of hematopoietic stem cells (HSC) in vitro, so as provide technical support for ultra long distance transport of HSC collected from unrelated donors. METHODS: Peripheral blood hematopoietic stem cells (PBHSC) were treated by different methods according to various groups, then stored at 4 ℃ in the refrigerator. The percentage of CD34+ cells, relative cell activity, relative cell proliferation rate, relative colony-forming rate, oxygen fraction and intracellular reactive oxygen species (ROS) were detected at 0, 24, 48 and 72 h after storage of PBHSC respectively. RESULTS: The percentage of CD34+ cells during 72 h storage did not altered. Along with the prolonging of storage time, the relative cell activity, relative cell proliferation rate and relative colony-forming rate gradually decreased in untreated PBHSC(control group), the related coefficients were -0.796, -0.883 and -0.815 respectively. Plasma dilution, antioxidants and oxygenation could improve the relative cell activity and relative cell proliferation rate, but oxygenation could decrease the relative colony-forming rate of PBHSC. The combination of 2 or 3 factors showed stronger protection effects on PBHSC. The intracellular level of ROS decreased gradually with the prolonging of storage time. Oxygenation of PBHSC could increase oxygen fraction, and also increase the intracellular level of ROS at the same time. The addition of antioxidants could reduce the level of ROS. CONCLUSION: The percentage of CD34+ cells can not serve as the indicator of PBHSC vitality. Plasma dilution, oxygenation and antioxidants can increase the survival and viability of PBHSC, but oxygenation can increase the intracellular ROS level and impair colony-forming ability of PBHSC. The combination of multiple factors can maintain the vitality of PBHSC better.

Prognostic Significance of Blood, Serum, and Ascites Parameters in Patients with Malignant Peritoneal Mesothelioma or Peritoneal Carcinomatosis.

To determine effects of the biochemical and cytological properties of blood, serum, and ascites on survival of patients with malignant peritoneal effusion (MPeE), including malignant peritoneal mesothelioma (MPeM) and peritoneal carcinomatosis (PC), we conducted a retrospective study of patients with MPeE and healthy controls. Potential prognostic factors were identified as follows: age, sex, blood neutrophil-to-lymphocyte ratio (NLR), serum parameters, ascites parameters, serum-ascites albumin gradient, and the ascites-serum LDH ratio. Compared to those of the control group, serum albumin levels were significantly lower, and the NLR and serum LDH levels were significantly higher in the MPeE group. Overall survival (OS) was longer in patients with MPeM compared to that in patients with PC. Compared with patients in the MPeM, patients with PC had higher NLRs, ascites glucose levels, serum-ascites albumin gradients, and serum LDH levels. In contrast, their ascites albumin levels and ascites-serum LDH ratios were lower. Univariate analyses indicated that the NLR, serum LDH levels, ascites LDH levels, ascites coenocyte levels, and the ascites coenocyte-to-monocyte ratios affected the OS. Multivariate analyses identified only serum and ascites LDH levels as independent prognostic factors.

Pretreatment Controlling Nutritional Status Score and Lactate Dehydrogenase as Predictive Markers of Survival in Patients with Malignant Peritoneal Mesothelioma.

OBJECTIVE: To investigate the relationships between the Controlling Nutritional Status (CONUT) score and ascites fluid lactate dehydrogenase (LDH) level, and prognosis in patients with malignant peritoneal mesothelioma (MPeM). METHODS: A total of 125 patients with MPeM were selected for the study using a pathological screening method. Once the diagnosis is established, before the treatment their clinical characteristics and nutritional evaluations were recorded including CONUT score and ascites LDH level. The associations between CONUT, ascites LDH, and other clinicopathological features including body mass index, asbestos exposure, pathological type, and treatment method were analyzed. Prognostic parameters predicting overall survival (OS) were analyzed by Cox regression. RESULTS: High CONUT score, high ascites LDH level were positively associated with poor prognosis in patients with MPeM according to univariate analyses (P < 0.001, P < 0.001, respectively), and CONUT score and ascites LDH were independent predictors of a poor prognosis according to multivariate analysis. When the CONUT score is greater than 3 and the ascites LHD is greater than 474 IU/l, it indicates a poor prognosis. CONCLUSIONS: CONUT score and ascites LDH are important factors influencing the prognosis of MPeM patients and should thus be considered in clinical applications.

[Detection of ATP Level in CD4+ T Lymphocytes and Its Clinical Significance in Allogeneic Hematopoietic Stem Cell Transplantation Recipients].

OBJECTIVE: To explore the clinical value of detecting adenosine triphosphate (ATP) level in CD4+ T lymphocytes (Immuknow ATP) of patients on early stage after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The base-line ATP value in CD4+ T lymphocytes in cases of hematological malignancies and the ATP level in CD4+ T lymphocytes of acute leukemia patients before allo-HSCT were detected. Allo-HSCT recipients were devided into 3 groups with different level of immunereactivity according to ATP concentraiton in month 3 (day 90±5) after allo-HSCT. The clinical characteristics of patients in 3 groups were analyzed. RESULTS: The mass concentration of Immuknow ATP in 15 cases of hematological malignancies before allo-HSCT ranged from 56.21-435.71 ng/ml, with a mean of 203.98±112.72 ng/ml. The ATP level in 46 cases after allo-HSCT ranged from 1.69-333.09 ng/ml, with a median of 41.96 ng/ml. Both 91.26 ng/ml (mean-SD) and 316.70 ng/ml (mean+SD) were used as cutoff, and 36 allo-HSCT recipients (78.3%) were assigned to low immunereactivity group, 8 recipients (17.4%) to middle group and 2 recipients (4.3%) to high group. The incidence of infection in low immunereactivity group was significantly higher than that in middle immunereactivity group (86.1% vs 50.0%)(P=0.022), and also significantly higher than that in high immunereactivity group (86.1% vs 0%)(P=0.002). There were no statistical differences in the incidences of severe infection among 3 groups. The incidence of grade II or higher acute graft versus host disease (aGVHD) in high immunereactivity group was superior to that in low immunereactivity group statistically (100% vs 13.9%)(P=0.002). Immune-mediated organ injury occurred more frequently in high immunereactivity group as compared with low and middle immunereactivity groups (100% vs 0% and vs 0%)(P=0.000; P=0.002). There were no significant differences in relapse rates of leukemia among 3 groups. The percentage of patients with increased trough blood concentration of cyclosporine A(CsA) was not significantly different among 3 groups (P=0.720). CONCLUSION: Detection of ATP level in CD4+ T lymphocytes on early stage after allo-HSCT possesses clinical significance for predicting infection, severity at aGVHD and immune-mediated organ injury.

CT differentiation of malignant peritoneal mesothelioma and tuberculous peritonitis.

OBJECTIVE: The aim of this study was to determine which computed tomography (CT) findings were useful in differentiating malignant peritoneal mesothelioma (MPM) and tuberculous peritonitis (TBP). METHODS: CT scans performed in 53 patients with MPM and 27 patients with TBP confirmed by pathology were retrospectively reviewed. The CT findings were evaluated for the morphologic appearance of ascites, peritoneum, mesenterium and omentum involvement, enlarged lymph nodes, solid abdominal viscera infiltration and metastases, and thoracic changes. The Pearson χ (2) test was used to compare differences between groups. RESULTS: Patients in both groups displayed a high proportion of peritoneum and mesenterium thickening. However, there were no obvious differences observed for ascites or swollen lymph nodes. There were significant differences in the following aspects between the two groups: (1) smooth peritoneal thickening was more frequent in patients with TBP, while irregular thickening was more frequently observed in patients with MPM; (2) caked omentum stratification was more common in patients with MPM; (3) mesentery involvement was less commonly observed in patients with TBP; (4) abdominal viscera infiltration and pleural plaques were more common in patients with MPM (46/53 and 48/53, respectively) than in those with TBP (0/27 and 0/27); and (5) more patients in the TBP group (14/27) displayed pleural effusion, and extraperitoneal tuberculosis was more common in patients with TBP (20/27). CONCLUSION: Although most CT findings analyzed are observed in both diseases, each disease has its own several unique characteristics. Therefore, using a combination of CT findings may increase our ability to distinguish TBP from MPM.

CT differentiation of diffuse malignant peritoneal mesothelioma and peritoneal carcinomatosis.

BACKGROUND AND AIM: Diffuse malignant peritoneal mesothelioma (DMPM) and peritoneal carcinomatosis (PC) have similar imaging in computer tomography (CT). We aimed to distinguish them. METHODS: Computer tomography findings were evaluated in 48 DMPM and 47 PC for the peritoneal, mesenteric, omentum, lymph nodes, viscera infiltration, ascites and pleural plaques. RESULTS: Two groups had no difference in terms of thickness, clinical manifestation, diameter of lymph nodes, ascites, and viscera infiltration. But they showed differences in the following: Ratio of asbestos exposure in DMPM group was higher. Smooth and irregular peritoneal thickening were more seen in DMPM group; peritoneal nodules were more commonly detected in PC group. Forty-eight cases of peritoneum in DMPM showed mild enhanced, while 14 patients in PC showed severe enhanced. Nodular type of omentum was more common in PC group than in DMPM group; omental cake was more commonly detected in DMPM group. Mesentery involvement was more commonly seen in DMPM group. Location of enlarged lymph nodes in cardiophrenic region was more frequently identified in DMPM, whereas location of enlarged lymph nodes in retroperitoneal region was more frequently identified in PC. Lymph nodes fusion was more frequently visualized in PC. Fixation of the intestinal wall was more common in DMPM. Pleural plaque was more common in DMPM. PC had distant metastasis except primary foci and peritoneum. In PC, tumor origins were ovary in 10, digestive system in 21, breast in one. CONCLUSION: Using a combination of CT findings may increase our ability to distinguish PC from DMPM.

[Biological characteristics of wharton's jelly derived mesenchymal stem cells after cryopreservation].

Aim of this study was to explore the effects of cryopreservation on biological characteristics of wharton's jelly derived mesenchymal stem cells (WJ-MSC), and to provide experimental evidence for clinical applications and the establishment of WJ-MSC bank. Primary WJ-MSC were produced by umbilical cord tissue culture in vitro. Fifth passage of WJ-MSC acquired by continuous cell culture were mixed with cryoprotectants, frozen in -80°C refrigerator and stored in liquid nitrogen. After the cryopreserved WJ-MSC were thawed, the first passage of WJ-MSC was obtained through cell culture and was taken as the 1st preserved passage (PP1). Thus, PP2-PP15 WJ-MSC were obtained by continuous cell subculture. The 1st control passage (CP1) to 15th passage (CP15) represented the 6th passage to 21st passage WJ-MSC acquired by subculturing in non-cryopreserved group. The biological characteristics of WJ-MSC from cryopreserved and control group, including the recovery rate of nucleated cells, trypan blue exclusion, CCK-8 activity, cell apoptosis, cell adherence, proliferation index, cell surface antigen, cell cycle and the capacities of induced differentiation into adipocyte, osteoblast and neuron, were detected and compared. The results indicated that the recovery rate of nucleated cells of cryopreserved WJ-MSC was 98.2%, trypan blue exclusion rate was 94.3%, CCK-8 activity was 91.4%, apoptotic rate was 3.9%, and the adherence rate was 92.6%. There was a statistically significant difference in proliferation index between PP1 and CP1 (P < 0.05), but there were no statistically significant differences between PP2-PP15 and their corresponding controls. The subculture cells highly expressed CD29, CD44, CD71, CD73, CD90, CD105, CD166 and HLA-ABC, and lowly expressed CD34, CD45 and HLA-DR. The expressions of above-mentioned surface antigens were not different statistically between two groups. The proliferation latency and logarithm proliferation of the subculture cells between two groups were also not different. After induced differentiation into adipocyte, osteoblast and neuron, the staining with oil red O, alkaline phosphatase and neuron-specific enolase was performed respectively, and the positive degrees were not clearly different macroscopically between two groups. Relatively high levels of triglyceride, alkaline phosphatase, and neuron-specific enolase in relevant cells could be detected, but had no significant differences between two groups. It is concluded that some WJ-MSC (< 10%) are damaged after cryopreservation, and the biological characteristics of WJ-MSC in cryopreservation group keep constant, as compared with that in non-cryopreservation group.

[Morphologic diagnosis and onset characteristics of acute leukemia: a retrospective analysis of 233 cases in 10 years].

The objective of this study was to evaluate the value of morphologic diagnosis for acute leukemia (AL), to explore the relation of morphologic diagnosis with immunology, cytogenetics and molecular biology diagnosis of AL and to analyze the onset characteristics of AL in 10 years. The samples of bone marrow and peripheral blood from 233 newly diagnosed cases of AL were collected during 2001-2011 years; the morphologic examination and immunologic, cytogenetic and molecular biologic examination (ICM) were carried out, the consistency of morphologic diagnosis with ICM diagnosis was compared, the onset characteristics of AL was analyzed. The results showed that: (1) the consistent rate of immunology, cytogenetics, molecular biology diagnosis with morphologic diagnosis was 84.3%. The order of consistent rat was AUL, M0 < M1 < HAL < M4 < M2 < M3 < M5 < ALL < M6, M7, AP; (2) Misdiagnosis always occurred among AUL, M0, M1, ALL and HAL or among M2a, M3v, M4 and M5. (3) In 233 cases, the highest ratio of blast was observed in M1 (92.5%), while the lowest ratio of blast was observed in M2 (49.5%). (4) AL occurred more frequently in males than that in female (147:86). (5) AL occurred in patients aged from 1 to 88 years. The median age was 41.5 for AUL, 40.8 for M0, 43.4 for M1, 46.3 for M2, 33.8 for M3, 42.6 for M4, 48.8 for M5, 77.3 for M6, 2.5 for M7, 65.0 for AP, 29.1 for ALL and 40.3 for HAL. (6) The number of patients in the later five years (139 cases) was significantly greater than that in the first five years (94 cases), especially the patients with M1, M2, M3, M4, and M5. It is concluded that morphologic diagnosis has important clinical value in the MICM diagnosis of AL. Attaching importance to the confusing cell morphology and onset characteristics of AL can improve the diagnostic accuracy.

[Influence of mesenchymal stem cells on proliferation of HL-60].

The objective of this study was to explore the effect of mesenchymal stem cells (MSC) on HL-60 proliferation and its molecular mechanism. HL-60 cells co-cultivated with MSC were used as experiment group, and the cells cultivated solely were taken as control group. HL-60 cells in the two groups were counted at different time. The time-quantity curve was drawn. The cell cycle and apoptosis ratio of HL-60 cells were compared between the two groups and expressions of Survivin and Bcl-2 protein were detected by flow cytometry. The results showed that HL-60 cells cultivated with MSC were obviously inhibited, especially on day 5 and 7 (P < 0.01). HL-60 cells were distributed on the phase of G(0)/G(1) [control group (47.0 ± 9.0)% vs experiment group (70.0 ± 16.0)%, P = 0.003], and apoptotic peak appeared. Both of Survivin and Bcl-2 protein expressions in HL-60 cells decreased [Bcl-2 protein in control group (63.0 ± 9.1)% vs experiment group (50.0 ± 14.1)%, P = 0.045; Survivin in control group (94.0 ± 9.3)% vs experiment group (77.0 ± 11.8)%, P = 0.006]. It is concluded that the MSC can inhibit HL-60 cell proliferation, and promote HL-60 cell apoptosis.

Effectiveness of human mesenchymal stem cells derived from bone marrow cryopreserved for 23-25 years.

OBJECTIVE: To evaluate long-term cryopreserved human bone marrow cells (BMCs) as a source of functional mesenchymal stem cells (MSCs). METHODS: Samples of human BMCs that were cryopreserved for 23-25 years (n=20) were thawed to obtain an initial culture and a primary culture (P(0)) that was propagated through five passages (P(1)-P(5)) to obtain MSCs. Freshly collected human bone marrow samples (n=20) were used as controls for comparison of efficiency of recovery and growth characteristics of MSCs. P(3) cultures were tested for their capacity to differentiate into osteoblasts, adipocytes, and neuronal cells. Appropriate staining, immunohistochemical and biochemical methods were employed to ascertain cell type identities at different stages of culturing. RESULTS: In the initial culture, the cell adherence rate of the cryopreserved cells was significantly lower than that of controls (19.7% vs. 38.2%, p<0.05) while the relative rate of recovery of MSCs was only 48.5±8.6% in P(0). At the end of P(3), fibroblast-like cells accounted for about 95% of cells in both cryopreserved and control groups (p>0.05). These cells were positive for essential MSC surface molecules (CD90, CD105, CD166, CD44, CD29, CD71, CD73) and negative for haematopoietic and endothelial cell markers (CD45, CD34, HLA-DR). The cell growth and cell cycle patterns were similar for both groups. MSCs at P(3) from both groups had similar capacities to differentiate in vitro into osteoblasts, adipocytes, and neuronal cells. CONCLUSION: Using the methods described here, long-term (23-25 years) cryopreserved human BMCs can be successfully cultivated to obtain MSCs that have good differentiation capabilities.

Noninvasive evaluation of renal oxygenation in diabetic nephropathy by BOLD-MRI.

PURPOSE: To evaluate the renal oxygenation in type 2 diabetes by blood oxygenation level dependent magnetic resonance imaging (BOLD-MRI). MATERIALS AND METHODS: Forty-eight patients with type 2 diabetes and 67 healthy controls were recruited. All patients were further divided into four subgroups based on renal functional level. Bilateral renal cortical R2* (CR2*) and medullary R2* (MR2*) values were extracted and quantified on BOLD-MRI, then R2* ratio between medulla and cortex (MCR) was calculated. CR2*, MR2* and MCR were compared among the groups separately. The relationships were analyzed between R2* values and clinical index of renal function. RESULTS: Compared with controls, MR2* and CR2* in diabetes were significantly increased. The positive relationship was found between MR2* and estimated glomerular filtration rate (eGFR), and CR2* was negatively correlated with eGFR. Interestingly, the MCR increased in early stage of diabetes and decreased along with the aggravation of diabetic nephropathy (DN). CONCLUSION: BOLD-MRI can non-invasively detect and assess the renal hypoxia in diabetes. Our findings suggest that hypoxia in medulla is more apparent and earlier than in cortex. During the progression of DN, a reversion of corticomedullary oxygenation gradient can be detected, thus, MCR would be adopted to suppose the progression and prognosis of DN.

[The effects of thrombopoietin on the fibrogenesis of bone marrow stromal cells in absence of megakaryocytes].

OBJECTIVE: In order to investigate whether or not thrombopoietin (TPO) could promote the fibrogenesis of bone marrow stromal cells in absence of megakaryocytes (MKs). METHODS: Improved dexter culture system with various TPO concentrations was used for ex vivo culture of bone marrow stromal cells. Relative proliferation index, the expressions of fibronectin, laminin and type IV collagen, and the systhesis of type III procollagen were detected at different time points during culture process. RESULTS: TPO stimulated the proliferation of bone marrow stromal cells. Relative proliferation index of the stromal cells increased with the TPO concentration increasing, and was not related to the exposure time. The expressions of fibronectin, laminin, and type IV collagen appeared stronger in the TPO groups than those in the control group. But the expressions of these molecules were not dependent upon the culture time. TPO could accelerate the synthesis of type III procollagen in bone marrow stromal cells, and this acceleration was unrelated to the TPO concentration. CONCLUSION: These findings suggested that TPO could stimulate the stromal cells with a consequence of increased syntheses and secretions of the extracellular matrix and collagen in absence of MKs. In other words, TPO could promote the fibrogenesis of bone marrow stromal cells without the existence of MKs.

[Non-invasive assessment of renal oxygenation with BOLD MRI in patients with type 2 diabetes].

OBJECTIVE: To evaluate renal oxygenation in patients with type 2 diabetes using blood oxygenation level-dependent magnetic resonance imaging (BOLD MRI). METHODS: The R2* values of cortex and medulla as well as cortical thickness were measured in 25 patients with type 2 diabetes and 30 normal controls. The associations between R2* values and clinical parameters were analyzed. RESULTS: The medulla R2* value of the patients with diabetes was significantly higher than that of the normal controls. No significant difference in cortical thickness was found between the patients with diabetes and the normal controls. The medulla R2* value increased with eGFR in the patients with diabetes. CONCLUSION: BOLD MRI is a non-invasive and efficient method to assess the oxygenation in different regions of kidney. Abnormality of kidney in diabetic patients can be detected earlier by BOLD MRI than traditional imaging.

[In vitro activity of human bone marrow cells after cryopreservation in liquid nitrogen for 21 - 25 years].

The aim of this study was to investigate the best method to preserve human bone marrow cells and the effectiveness of long term cryopreservation at -80 degrees C. The human bone marrow cells in 20 samples were firstly frozen by a programmed freezer or -80 degrees C refrigerator, and then were preserved in liquid nitrogen with DMSO-AuP (10% dimethylsulfonamide, 10% autologous plasma) or DMSO-HES-HuA (5% dimethylsulfonamide, 6% hydroxyethyl starch, 4% human serum albumin) as cryoprotectant for 21 to 25 years. They were thawed in 38 degrees C. The cell sample frozen in -80 degrees C refrigerator was frozen at a low frozen speed of 1 degrees C/min which was the same as the programmed freezer before -30 degrees C. Before detection the bone marrow cells were taken from liquid nitrogen and were thawed in 38 degrees C, then the suspension of bone marrow cells was prepared for detection. The cell morphology and recovery rate of erythrocytes, nucleocytes and platelets; the recovery rate of hematopoietic stem progenitors cells, as well as mesenchymal stem cells were determined. The results showed that the protective effectiveness of DMSO-HES-HuA was better than DMSO-AuP. The mature erythrocytes were destroyed lightly [(3.5 +/- 1.5)% versus (12.6 +/- 4.8)%], the hemolysis rate was lower [(3.3 +/- 1.6)% versus (23.1 +/- 5.1)%]. Osmotic fragility of erythrocytes in the former was not changed, but was dropped in the latter. The recovery rates of red cell, platelet, granulocyte-macrophage colony forming units and long term culture-initiating cells were higher in the former than that in the latter [(96.1 +/- 1.8)%, (70.0 +/- 9.5)%, (49.2 +/- 10.9)%, (54.2 +/- 13.8)% versus (76.3 +/- 5.6)%, (52.7 +/- 8.1)%, (43.5 +/- 12.3)%, (47.2 +/- 13.6)% respectively]. With each kind of cryoprotectant or frozen method, the frozen MSC could keep the original growth properties. With the same cryoprotectant and different frozen method, the cryopreservative effectiveness was not different. The influence of the cryoprotectant prescriptions and the frozen methods on the cryopreservative effectiveness was little. It is concluded that the human bone marrow cells with DMSO-AuP or DMSO-HES-HuA as cryoprotectant, frozen by a programmed freezer or -80 degrees C refrigerator, could be then preserved in liquid nitrogen for long time. When the preserving time was as long as 21 to 25 years, the morphology, the recovery rate and the activity of various kinds of cells were still good. The method of freezing by -80 degrees C refrigerator with 5% DMSO-6% HES-4% HuA and preserving in liquid nitrogen would be convenient, cheap and easily-manipulated for preservation of the human bone marrow cells.

[Influences of bone marrow mesenchymal stem cells in patients with acute myeloid leukemia and non-leukemia on HL-60 cells -- a comparison study].

This study was aimed to compare the influences of bone marrow mesenchymal stem cells (BMMSCs) from patients with acute myeloid leukemia (AML), AML patients with complete remission (CR) and non-leukemia patients on HL-60 cells. The HL-60 cells were divided into three groups: group of co-cultivation with BMMSCs of AML patients, group of co-cultivation with BMMSCs of AML patients with CR and group of co-cultivation with BMMSCs of non-leukemia patients. The count of HL-60 cells, the CD11b and survivin expression of HL-60 cells, the cell cycle distribution of the HL-60 cells in 3 groups were compared by flow cytometry, the morphology and differentiation rate of HL-60 cells in 3 groups were observed and compared by microscopy. The results showed that there were no differences in HL-60 cell count at five and seven days, in HL-60 distribution at the G(0)/G(1) phase, in survivin and CD 11b expressions in 3 groups. All cells of 3 groups began to mature, and the differentiation rates in 3 groups were 18.0 +/- 3, 17.0 +/- 1.3 and 19.0 +/- 2.0 respectively, therefore there were no significant differences between the 3 groups (p = 0.23). It is concluded that there is no influence of BMMSCs in 3 groups on the proliferation and differentiation of HL-60 cells.

[Comparison of biological characteristics of bone marrow mesenchymal stem cells in acute myeloid leukemia with those from non-leukemia].

This study was aimed to compare partial biological characteristics of bone marrow mesenchymal stem cells (BMMSCs) in AML patients, AML patients with complete remission (CR) and non-leukemia patients. The bone marrow (BM) MSCs were divided into 3 groups: group of MSCs from AML patients, group of MSCs from AML patients with CR, group of MSCs from non-leukemia patients. The morphologic features of MSCs were observed by light microscopy; CFU-F numbers of MSCs were counted after Wright-Giemsa staining in situ; the fusion times of MSCs were determined; the growth curves of MSCs were drawn by counting cell numbers; the immunophenotypes and cell cycle of MSCs were detected by flow cytometry; the DI values of MSCs were calculated. The results showed that the morphologic features of MSCs in 3 groups did not display difference; there was significant difference (p < 0.01) of CFU-F numbers in 3 groups, while CFU-F number of MSCs in AML group was minimal; there was significant difference of MSC fusion time in 3 groups, while fusion time of MSCs in AML group was most long; the growth curves of MSCs in 3 groups were similar; MSCs in 3 groups highly expressed CD105 and CD106, but not expressed CD45; the cell distribution ratios at phase of G(0)/G(1) for MSCs in 3 groups were 89.9 +/- 4.0%, 90.2 +/- 3.0% and 91.0 +/- 3.0% respectively; the DI values of MSCs in 3 groups were between 0.9 and 1.1. It is concluded that no significant difference of biological characteristics of the second generations of MSCs is found between those in leukemia and non-leukemia patients.

[CT findings on renal damage in patients with liver cirrhosis].

OBJECTIVE: To use CT to identify renal damage in patients with liver cirrhosis. METHODS: Sixty patients with liver cirrhosis confirmed by clinical, laboratory and imaging examinations and 20 patients without liver and kidney diseases (control group) were recruited in this study. The participants underwent double-stage enhanced scanning. The thickness of renal cortex (C) and parenchyma (P), and enhancement of renal cortex and medulla on cortical phase were measured. The association between renal damage and Child-Pugh grades, ascites capacities, liver cirrhosis types, opened portal collateral vessels, and opened collateral vessels between splenic venous and renal venous in the patients with liver cirrhosis were analysed. RESULTS: The patients with liver cirrhosis had lower C/P and enhancement of cortex on cortical phase and greater enhancement of medulla than the controls (P < 0.05). The enhancement of cortex decreased with the increase of Child-Pugh grade, opened portal collateral vessels and opened collateral vessels between splenic venous and renal venous (P < 0.05), but the differences of enhancement of medulla and C/P had no statistical significance (P > 0.05). No differences in the measured indicators were found between patients with different capacities of ascites and patients with different types of liver cirrhosis (P > 0.05). CONCLUSION: The CT findings on renal damage in patients with liver cirrhosis include decrease in C/P and enhancement of cortex on cortical phase, and increase in enhancement of medulla on cortical phase. The enhancement of cortex on cortical phase is the most important change, which is associated with Child-Pugh grades, opened portal collateral vessels and opened collateral vessels between splenic venous and renal venous.

[Effects of recombinant human thrombopoietin on stromal cells in culture in vitro].

This study was aimed to investigate whether the thrombopoietin (rhTPO) may facilitate myelofibrosis or not. The modified Dexter culture system with various concentrations of rhTPO was used to culture the stromal cells in vitro; the proliferative activity of cells was detected by MTT method; the morphologic changes were observed by light and scanning electron microscopy; the staining changes of ALP, PAS, AS-D NCE and IV type collagen were observed by cytochemistry method; the changes of fibronectin, laminin and IV type collagen were assayed by immunohistochemistry method; the cell surface antigens were assayed by flow cytometry. The results indicated that rhTPO could promote the proliferation of stromal cells which was related to the concentrations of rhTPO. Proliferative activity of stromal cells increased with increasing of rhTPO concentration, and was not related to the exposure time. On day 3 stromal cells adhered to the wall, and became oval. On day 7 stromal cells turned to fusiform and scattered dispersively. On day 12 to 14 these cells ranged cyclically and became long fusiform. Cells covered 70%-80% area of bottle bottom at that time. By day 16 to 18 these cells covered more than 90% area of bottom and ranged cyclically. They displayed the same shape as fibroblasts. By light microscopy with Wrights-Giemsa staining, fibroblasts predominated morphologically, few macrophages, endothelial cells and adipose cells were found. There were no significant differences between experimental group and control group. On day 14 to 42 the adherent cells were positive with PAS staining, poorly positive with ALP and naphthol AS-D chloroacetate esterase (AS-D NCE) staining, and the difference in cytochemistry was not significant between two groups. When these cells were dyed with Masson's trichrome and Gomori's staining, neither collagen fibers nor reticular fibers were positive, but fibronectin, laminin, and collagen type IV appeared positive stronger in experimental group than those in control. The expressions of these molecules were not dependent on culture time. By scanning electron microscopy microvilli and fibers on cell surface appeared more and more, monolayer cells evolved into multilayer cells, and newly-formed fibroblasts appeared gradually as culture time prolonged. These alterations were not different among various groups. The expressions of CD34, CD45, CD105, CD106, and CD166 were not affected obviously by rhTPO. It is concluded that rhTPO had no effects on histochemical properties of stromal cells. Fiber staining and scanning electron microscopic examinations revealed that rhTPO can not facilitate fiber formation of stromal cells. But rhTPO may be able to augment the expressions of fibronectin, laminin and collagen type IV of stromal cells. Therefore it is still necessary to follow up the patients for a long time, who have received rhTPO therapy clinically.