Effects of H2SO4, GA3, and cold stratification on the water content, coat composition, and dormancy release of Tilia miqueliana seeds.

Introduction: Tilia miqueliana is an endemic species whose population is declining. The permeability barrier and mechanical constraint of the pericarp (seed coat) are important causes of its seed dormancy. Although there has been considerable research on this subject, questions remain regarding how the permeability barrier and mechanical constraint of the seed coat are eliminated during dormancy release and how water enters the seed. Therefore, protecting the species by improving its germination/dormancy breaking in the laboratory is urgent. Methods: In this study, the changes in the cellular structure, mechanical properties, and components of the Tilia miqueliana seed coat after an H2SO4-gibberellic acid (GA3) treatment were analyzed during dormancy release. Various analyses (e.g., magnetic resonance imaging, scanning electron microscopy, and paraffin section detection) revealed the water gap and water channel. Results: The H2SO4 treatment eliminated the blockage at the micropyle and hilum of the seeds. Water entered the seeds through the water gap (micropyle) rather than through the hilum or seed coat, after which it dispersed along the radicle, hypocotyl, and cotyledon to the endosperm. During the cold stratification period, the cellular structure was damaged and an increasing number of holes appeared on the inner and outer surfaces of the seed coat. Vickers hardness tests showed that GA3 decreased the seed coat hardness. Additionally, the seed coat lignin and total phenol contents continuously decreased during the cold stratification period. Notably, the Liquid chromatography-mass spectrometry (LC-MS) analysis of the seed coat detected polyethylene glycol (osmoregulator), which may have destabilized the water potential balance inside and outside the seed and increased the water content to levels required for germination, ultimately accelerating seed dormancy release. Discussion: This sophisticated and multi-level study reveals how H2SO4 and GA3 eliminate the permeability barrier and mechanical constraints of the seed coat during dormancy release of Tilia miqueliana seeds. This will be beneficial to artificially assist the natural regeneration and population expansion of Tilia miqueliana.

Toward Energy-Efficient Routing of Multiple AGVs with Multi-Agent Reinforcement Learning.

This paper presents a multi-agent reinforcement learning (MARL) algorithm to address the scheduling and routing problems of multiple automated guided vehicles (AGVs), with the goal of minimizing overall energy consumption. The proposed algorithm is developed based on the multi-agent deep deterministic policy gradient (MADDPG) algorithm, with modifications made to the action and state space to fit the setting of AGV activities. While previous studies overlooked the energy efficiency of AGVs, this paper develops a well-designed reward function that helps to optimize the overall energy consumption required to fulfill all tasks. Moreover, we incorporate the e-greedy exploration strategy into the proposed algorithm to balance exploration and exploitation during training, which helps it converge faster and achieve better performance. The proposed MARL algorithm is equipped with carefully selected parameters that aid in avoiding obstacles, speeding up path planning, and achieving minimal energy consumption. To demonstrate the effectiveness of the proposed algorithm, three types of numerical experiments including the ϵ-greedy MADDPG, MADDPG, and Q-Learning methods were conducted. The results show that the proposed algorithm can effectively solve the multi-AGV task assignment and path planning problems, and the energy consumption results show that the planned routes can effectively improve energy efficiency.

Analysis of the Effect of Nine Consecutive Years' Intensive Management and Number of Times Achieving the Target Control on Endpoint Events in T2DM Patients in Sanlitun Community Health Service Center in Beijing.

OBJECTIVE: To investigate the effect of intensive management and achieving the target control more than 3 times on endpoint events during 9 consecutive years' annual assessment in type 2 diabetes (T2DM) patients in the Sanlitun Community Health Service Center in Beijing, including blood glucose, blood pressure, lipids profiles, and the joint target control. METHODS: In Beijing Community Diabetes Study (BCDS), 224 patients with T2DM from the Sanlitun Community Health Service Center were enrolled in 2008. All patients were randomly assigned to the intensive management group (n = 113) and the standard management group (n = 113) and the standard management group (. RESULTS: During the nine-year follow-up, the abscission number was 35 (14.29%), among which 14 (12.39%) was in the intensive management group and 21 (18.92%) was in the standard management group. The incidence of diabetic retinopathy (6 cases, 5.41%) and diabetic nephropathy (13 cases, 11.71%) in the standard management group was significantly higher than that in the intensive management group (1 case, 0.88%; 5 cases, 4.42%), respectively (P < 0.05). However, there were no significant differences on the other endpoint events between the two groups (P < 0.05). However, there were no significant differences on the other endpoint events between the two groups (P < 0.05). However, there were no significant differences on the other endpoint events between the two groups (P < 0.05). However, there were no significant differences on the other endpoint events between the two groups (P < 0.05). However, there were no significant differences on the other endpoint events between the two groups (. CONCLUSIONS: The intensive management can effectively reduce the occurrence of microvascular complications. The incidence of all-cause death and the other endpoint events decreased in T2DM patients who achieved the joint target control more than 3 times during the nine-year management, which improved survival time and life quality. This trial is registered with ChiCTR-TRC-13003978 and ChiCTR-OOC-15006090.

Endothelial progenitor cells in peripheral blood may serve as a biological marker to predict severe acute pancreatitis.

AIM: To investigate the significance of endothelial progenitor cells (EPCs) in predicting severe acute pancreatitis (SAP). METHODS: We recruited 71 patients with acute pancreatitis (AP) and excluded 11 of them; finally, cases of mild acute pancreatitis (MAP) (n = 30) and SAP (n = 30), and healthy volunteers (n = 20) were internalized to investigate levels of EPCs, C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), fibrinogen (FIB) and white blood cells (WBC) in peripheral blood. RESULTS: The levels of TNF-α, WBC, FIB and CRP were higher both in SAP and MAP cases than in healthy volunteers (P < 0.05, all). Interestingly, the level of EPCs was higher in SAP than MAP (1.63% ± 1.47% vs 6.61% ± 4.28%, P < 0.01), but there was no significant difference between the MAP cases and healthy volunteers (1.63% ± 1.47% vs 0.55% ± 0.54%, P > 0.05). Receiver operating characteristics curve (ROC) showed that EPCs, TNF-α, CRP and FIB were significantly associated with SAP, especially EPCs and CRP were optimal predictive markers of SAP. When the cut-off point for EPCs and CRP were 2.26% and 5.94 mg/dL, the sensitivities were 90.0% and 73.3%, and the specificities were 83.3% and 96.7%. Although, CRP had the highest specificity, and EPCs had the highest sensitivity and highest area under the curve value (0.93). CONCLUSION: Data suggest that EPCs may be a new biological marker in predicting SAP.

Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by Attenuated Salmonella in Rats.

The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 109 cfu), vehicle (TP, 1 × 109 cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm², which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm², respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm², which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.

High altitude increases the expression of hypoxia-inducible factor 1α and inducible nitric oxide synthase with intest-inal mucosal barrier failure in rats.

BACKGROUND: The failure of intestinal mucosal barrier may induce multiple organ dysfunction and systemic inflammatory response syndrome, but little work has been done on whether hypobaric hypoxia related to the failure of intestinal mucosal barrier. AIMS: To study the expression of hypoxia-inducible factor 1α (HIF-1α), inducible nitric oxide synthase (iNOS) and morphological changes of intestinal mucosa in albino rats at different altitude. METHODS: 30 male Wistar rats raised in plain for one month were randomly divided into 3 groups: Plain 500 m group (n=10), High-altitude (HA) 3842 m group (n=10) and HA4767 m group (n=10). Each group was delivered to different altitude area at the same shipping time and executed after 3 days' exposure to different altitude. Intestinal segments with the same location of all rats were removed for morphological analyses. Morphologic parameters (villous height, crypt depth, mucosal wall thickness and villous surface area) were measured by optical and scanning electron microscope. The expression of iNOS and HIF-1α were detected by immunohistochemistry. RESULTS: Morphological indexes in higher altitude groups were exacerbated obviously compared with those of lower altitude groups. While the expression of iNOS and HIF-1α in higher altitude groups were significantly increased than those of lower altitude groups. Linear correlation analysis showed that the expression of iNOS was positively correlated with that of HIF-1α. CONCLUSIONS: Hypobaric hypoxia increases the expression of HIF-1α and iNOS in intestinal mucosa, however exacerbates the mucous morphologic parameters with altitude increasing. HIF-1α may regulate the expression of iNOS and be involved in the damage of intestinal mucosa.

[Reconstruction of deep wounds in old patients with cutaneous malignant tumor after artificial dermis combined with negative pressure wound therapy].

OBJECTIVE: To evaluate the effects of artificial dermis combined with negative pressure wound therapy on repairing wound aft er resection of cutaneous malignant tumor in elderly. METHODS: The clinical data of 34 hospitalized patients with cutaneous malignant tumor from July, 2009 to February, 2014 were retrospectively analyzed. The patients received local enlarged excision, artificial dermis covered wounds and 12-16 days of negative pressure wound therapy plus a free skin graft transplant on the surface with an artificial dermis. Recovery and complication aft er operation were assessed. RESULTS: All wounds were restored successfully with good appearance. None severe infection happened. CONCLUSION: Artificial dermis combined with negative pressure wound therapy can repair wound efficiently aft er cutaneous malignant tumor resection was performed for old patients.

Activation of autophagy in rats with plateau stress-induced intestinal failure.

Hypobaric hypoxia may damage the intestinal mucosa, which may induce multiple organ dysfunction. However, little work has been done regarding whether high altitude hypoxia is associated with failure of the intestinal mucosal barrier. The aim of this study was to investigate the change of the autophagy after the intestinal failure in rats acutely exposed to plateau stress. Fifty Wistar rats were randomly divided into five groups: the plain group, plateau for 6 h, 12 h, 24 h, and 48 h (n = 10 in each group). The acute exposure to plateau was established at a simulated altitude of 4767 meters (m) in a decompression chamber. Intestinal injury was verified by light microscopy. The autophagosomes in the intestinal epithelial cells were observed by transmission electron microscopy (TEM). The protein expression of Beclin1 and LC3B in the intestinal epithelial cells were analyzed by immunohistochemistry. Compared with the plain group, acute exposure to plateau led to a time-dependent damage of the intestinal epithelium. The autophagosome was observed after the intestinal failure following acute exposure to high altitude for 6 h. The expression of Beclin1 and LC3B protein in the rats exposed to acute plateau for 6 h, 12 h, 24 h and 48 h were significantly higher than those in the plain group. The expression of autophagy also showed a significant increase in rats with intestinal failure following acute exposure to plateau stress.

Expression of breast cancer resistance protein in peripheral T cell subsets from HIV­1­infected patients with antiretroviral therapy.

The aim of the present study was to investigate the expression of breast cancer resistance protein (BCRP) in peripheral T cell subsets of human immunodeficiency virus 1 (HIV­1)­infected patients, and to analyze the association between the levels of BCRP expression and disease progression in HIV­1 infection. Peripheral blood mononuclear cells (PBMCs) were obtained from HIV­1­infected patients (n=118), including 92 patients with antiretroviral therapy (ART) and 26 patients without a history of ART. Control samples from 30 healthy donors were also analyzed. The expression levels of BCRP in T cells were evaluated by flow cytometry. A high inter­individual variability was observed in CD4+ and CD8+ T cells in the HIV­1­infected patients and healthy donors; however, the analyzed expression levels of BCRP were significantly higher in the HIV­1­infected group with ART than those in the group with no history of ART (P<0.01). Furthermore, the frequency of BCRP­expressing T cells was inversely correlated with CD4+ and CD8+ T cell counts in HIV­1­infected patients with ART. The results suggested that BCRP expression varied among HIV­1­infected patients and healthy donors but was significantly higher in HIV­1 patients undergoing ART. In conclusion, the present study suggested that overexpression of BCRP may be involved in disease progression of the HIV­1 infection and may participate in drug resistance to ART, thus contributing to the failure of highly active ART in HIV­1 therapeutics.

Distribution of endothelial progenitor cells in tissues from patients with gastric cancer.

It is accepted that endothelial progenitor cells (EPCs) are recruited into tumor sites and take part in the neovascularization of tumors. However, few articles have discussed the specific distribution of EPCs in vivo in tissues of gastric cancer patients. For this reason, the present study sought to elucidate EPC distribution in vivo in tissues of patients with gastric cancer. Fresh tumor tissues were collected from 26 newly diagnosed patients with histologically confirmed gastric cancer (mean age, 51 years; range, 21-81 years; 7 females, 19 males). All patients were treated surgically with curative intent. One portion of the fresh tissues was prepared for flow cytometric analysis and another was immediately snap frozen in liquid nitrogen and stored at -80°C for later use in quantitative polymerase chain reaction. The analysis was based on two groups of tissues, namely the cancer group and cancer-adjacent group. The presence of CD34/CD133 double-positive cells was determined in cancer-adjacent and cancer tissues by flow cytometry. The analysis revealed that the total number of EPCs in cancer tissue was slightly greater than the number in the cancer-adjacent tissue, but not to the point of statistical significance. The number of EPCs in cancer-adjacent and cancer tissues of patients with early-stage gastric cancer was higher than the EPC count in late-stage gastric cancer patients, and significant differences were identified in the number of EPCs in cancer tissue between patients of different tumor stages. Levels of cluster of differentiation (CD)34, CD133 and vascular endothelial growth factor receptor 2 were not significantly different in cancer-adjacent tissue compared with cancer tissue. These results suggest that cancer-adjacent and cancer tissue of gastric cancer patients may be used as a reference index in the clinical and pathological staging of tumors.

Expression levels of P-glycoprotein in peripheral blood CD8+ T lymphocytes from HIV-1-infected patients on antiretroviral therapy.

In this study, we aimed to measure P-glycoprotein (P-gp) expression in CD8(+) T lymphocytes of HIV-1-infected patients, to investigate how P-gp levels are affected by antiretroviral therapy (ART) in HIV-1 infection, and to assess the value of using P-gp expression to predict virologic response to ART. Peripheral blood mononuclear cells (PBMCs) were obtained from a cohort of HIV-1­infected patients in China: 140 patients on ART, and 49 ART-naïve patients. We also enrolled 24 healthy blood donors as the controls. The expression levels of P-gp in CD8(+) T cells of HIV-1-infected patients were evaluated by quantitative reverse transcription PCR, ELISA and flow cytometry. A high inter-individual variability was observed in the CD8(+) T cells of both HIV-1-infected patients and healthy donors; however, the expression levels of P-gp were significantly higher in the HIV-1-infected group on ART compared to the ART-naïve group. The relative proportion of P-gp(+)CD8(+) T cells inversely correlated with the blood CD4(+) T cell count in the HIV-1­infected patients on ART (r=-0.3343, P=0.0375). Groups of both good and poor responders showed significantly elevated levels of P-gp(+)CD8(+) T cells. The percentage of P-gp(+)CD8(+) T cells appeared to provide a sensitive estimate of the virologic response to ART compared to the CD4(+) T cell count. Our results suggest that P-gp expression varies among HIV-1­infected patients, but is significantly higher in HIV-1­infected patients on ART. The overexpression of P-gp is involved in ART initiation during HIV-1 infection, and P-gp(+)CD8(+) T cells may be an additional criterion for the evaluation of the antiretroviral response to ART.

Identification and clinical significance of circulating endothelial progenitor cells in gastric cancer.

CONTEXT: There are few reports of endothelial progenitor cells (EPCs) in peripheral blood have been found in patients with gastric cancer. OBJECTIVE: We quantified EPCs in the peripheral blood of patients with gastric cancer, with the expectation that this approach might lead to a new marker for the diagnosis of gastric cancer. METHODS: We enumerated CD34+/CD133+ EPCs in the peripheral blood of 145 subjects by use of flow cytometry. RESULTS AND CONCLUSION: The quantity of peripheral blood EPCs in patients with gastric cancer are correlated with patient's age. In addition, the number of peripheral blood EPCs in patients with gastric cancer increased with tumor node metastasis stage and histological differentiation of the cancers, and with the operative status of the patients.

[Construction of recombinant adenovirus vector pAdxsi-GFP-HIF containing hypoxia inducible factor gene and its expression in endothelial cells].

OBJECTIVE: To construct a recombinant adenovirus (pAdxsi-GFP-HIF) encoding human hypoxia inducible factor 1 α gene (HIF-1 α) and to express it in endothelial cells. METHODS: HIF-1 α gene was obtained from human lung cancer cell line A549, which was cultured in hypoxia condition, by RT-PCR. The HIF-1 α gene was subcloned into shuttle vector p Shuttle-CMV-EGFP at KpnI and BamHI sites. After identified with restriction enzymes, plasmid p Shuttle-GFP-HIF was linearized by digestion with restriction endonuclease I-CeuI and I-SceI, and subsequently cotransformed into E.coli DH5a with adenoviral backbone plasmid pAdxsi to make homologous recombination. After linearized by PacI, the homologous recombinant adenovirus plasmid was transfected into 293 cells to package and amplify. The recombinant adenovirus was infected with human umbilical vein endothelial cells (ECV304), and the expression level of HIF-1 α protein was evaluated by ELISA. RESULTS: The recombinant adenovirus vector containing HIF-1 α gene (pAdxsi-GFP-HIF) was successfully constructed and amplified with titer of 3.38 X 10(10) pfu/mL. The green fluorescence protein was detected under fluorescent microscope in ECV304 at 24h after transfection and with a stronger degree after 48h. The concentration of HIF-1 protein was (48.93 ±3.86)ng/mL in supernatant at 48 h after transfection. CONCLUSION: A recombinant adenovirus vector pAdxsi-GFP-HIF, encoding human hypoxia inducible factor 1 α gene, has been constructed in vitro and expressed successfully in ECV304 cells.

Inhibitory effects of the attenuated Salmonella typhimurium containing the IL-2 gene on hepatic tumors in mice.

To observe the inhibitory effects of an attenuated S. typhimurium strain carrying IL-2 gene (TPI) on hepatoma cell line (HepG2) and transplanted tumors in mice. TPI, TPG (an attenuated S. typhimurium strain carrying green fluorescent protein gene), and TP (an attenuated S. typhimurium strain) strains were transfected into HepG2 cells. At 48 h after transfecting, the transfection rate was 82.58 ± 1.74%. The expression level of IL-2 was (99.5 ± 12.2) ng/1 × 10(6) cells. Compared with TPG, TP, and normal mouse groups, the proportion of CD4(+) T and CD8(+) T cells in the blood from the TPI group was higher, the levels of IgM and IgG(1) were significantly increased, and the proliferation activity of splenic lymphocyte was significantly stronger. The transplanted tumor weight in the TPI group was significantly smaller than that in the other two groups. The infiltration of lymphocytes increased in the tumor from TPI group mice. TPI was effectively transfected into cancer cells, which expressed the protein of interest. Oral administration of TPI prolonged survival of mice transplanted with hepatoma cell tumours.

[Effects of glutamine enriched enteral feeding on immunoregulation in burn patients].

OBJECTIVE: To investigate the effects of glutamine enriched enteral feeding on immunoregulation in burn patients. METHODS: Twenty burn patients were randomly divided into enteral nutrition (EN) group and enteral immune nutrition (EIN) group, with 12 patients in each group. Patients in EN group received a standard enteral formula, while those in EIN group received an enteral formula enriched with glutamine after hospital admission. Nutritional support was continued for 10 days. Blood samples were obtained to determine plasma level of total protein (TP), albumin (ALB), prealbumin (PAB) and transferrin (TF) at 1, 4, 7, 10 post-burn days (PBD). At the same time the concentration of immunoglobulin (IgA, IgG and IgM) were determined, the percentage of CD3+, CD4+, CD8+ subpopulations of T lymphocytes, and the ratio of CD4+/CD8+ were determined by FCM. RESULTS: (1) There were no obvious difference of the plasma level of TP, ALB, TF, CD3+, IgM between the two groups at each time-point (P > 0.05). (2) The plasma PAB contents in EIN group were significant higher than that in EN group on 4 PBD [(90 +/- 14 vs 60 +/- 15) mg/L, P < 0.05], 7 PBD [(92 +/- 16 vs 64 +/- 13) mg/L, P < 0.05] and 10 PBD [(106 +/- 21 vs 72 +/-16) mg/L, P < 0.05]. (3) The percentage of CD4+ subpopulation and ratio of CD4+/CD8+ in EIN group were obviously higher than those in EN group on 7 PBD [CD4+ (55 +/- 5 vs 45 +/- 5)%, CD4+/CD8+ (1.92 +/- 0.31 vs 1.53 +/- 0.27)%, P < 0.05] and 10 PBD [CD4+ (56 +/- 5 vs 49 +/- 5)%, CD4+/CD8+ (2.36 +/- 0.36 vs 1.72 +/- 0.42), P < 0.05]. (4) The concentration of IgA and IgG in EIN group were markedly higher than that in EN group on 7 PBD [IgA (2.8 +/- 0.6 vs 2.2 +/- 0.5) g/L, IgG (12.1 +/- 1.3 vs 9.8 +/- 1.2) g/L, P < 0.05] and 10 PBD [IgA (3.1 +/- 0.6 vs 2.5 +/- 0.5) g/L, IgG (14.2 +/- 1.3 vs 10.4 +/- 1.3) g/L, P < 0.05]. CONCLUSION: These findings suggest that glutamine enriched enteral feeding can improve nutritional status by promoting the synthesis of IgA, IgG, and increasing the PAB concentration, and corrected immunologic dysfunction in burn patients.

[The influence of apoptosis of lymphocytes of Peyer's patches on the pathogenesis of gut barrier damage in severely scalded mice].

OBJECTIVE: To investigate the changes in cellular apoptosis of Peyer's patches in severely scalded mice, and to explore its role in the pathogenesis of gut barrier damage. METHODS: Forty BALB/c mice were randomly divided into normal control, 12 post-scald hour (12PSH), 24PSH and 72PSH groups, with 10 in each group. The mice in all PSH groups were inflicted with 20% TBSA full-thickness scald on the back. The mice in all the groups were sacrificed at different time points, and Peyer's patches were harvested from all the mice for HE staining, DNA gel electrophoresis, and flow cytometry ( FCM) examination with FITC conjugated Annexin-v and propidium iodide( PI) staining of cells. RESULTS: HE staining revealed that there were relatively abundant apoptotic cells scattering in Peyer's patches of scalded mice . DNA electrophoresis of Peyer's patches revealed typical " ladder" pattern at all indicated time points in scalded mice. Apoptotic percentage of detached Peyer's patches cells in control and scalded group were (4. 9+/-2. 1)% , (26.7+/-3. 1)% , (21.6 +/-4.0)% ,(12. 8 +/-2.0)% , respectively, and the percentage reached the peak at 12 PSH. CONCLUSION: Apoptosis is a principle modality of cell death of small intestinal Peyer's patches lymphocytes in severely scalded mice, and it might contribute to immunity barrier failure of intestinal wall after severe thermal injury.

[Changes in the calcitonin content after inhalation injury in dogs].

OBJECTIVE: To investigate the changes in the serum content of immunoreactive calcitonin (iCT) after burns or inhalation injury, and to explore its diagnostic significance. METHODS: Twenty-four dogs were randomized into 4 groups, i. e. A (n = 6, with moderate degree inhalation injury) , B ( n = 6, with severe inhalation injury), C (n = 6, with most severe inhalation injury) and D (n = 6, with severe burns) groups. The serum content of iCT and blood gas analysis before and after injury were determined at different time points. The degree of inhalation injury was determined with fibrobronchoscopic examination at 6 post-inhalation injury hour (PIH). RESULTS: (1) Fiber bronchoscopic examination showed that the degree of inhalation injury in each group was coincident with the anticipation. (2) The serum content of iCT in each group at 1 PIH was obviously higher than that before injury, and it was evidently higher in A, B and C groups than that in D group at 4 PIH. The peak value of iCT in group A at 24 PIH was (453+/-224) ng/L, and it increased gradually in B and C groups at 48 PIH. The serum content of iCT increased continually from 2 PIH on, and it reached (125+/-41) ng/L at 48 PIH. (3) Compared with PaO2 value before injury (109+/-8) mmHg, there was no obvious difference of the PaO, in A and D groups. PaO2 value in B and C group began to descend continually at 8 PIH (65+/-6) mmHg, and that in C group began to descend at 4 PIH (71+/-9) mmHg. PaCO2 value in C group began to increase at 24 PIH(52+/-11) mmHg when compared with that before injury(38+/-5 ) mmHg. CONCLUSION: The changes in the serum level of iCT within 8 PIH occurred much earlier than PaO2 and PaCO2, thus it has the same diagnostic significance as fibers bronchoscopic examination.

[Maturation regulation of dendritic cells pulsed with hepatocellular carcinoma cell soluble antigens].

OBJECTIVE: To research the maturation regulation of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) cell soluble antigens. METHODS: BCG HSP 70 was purified by SDS-PAGE electrophoresis and its biological activity was determined with ELISA. Phenotypes of DCs pulsed with antigens or with both antigens and BCG HSP 70 were analysed with flow cytometry. MTT assay was used to estimate the proliferation of self lymphocytes and the mixed lymphocyte reaction (MLR) of BCG HSP 70 primed DCs. RESULTS: The characteristics of DCs had changed after loaded with soluble antigens of HCC. There were about 10% DCs which had lost their specific markers. The expression levels of CD54, CD83, CD86 molecules and the stimulatory ability in allogeneic MLR decreased. However, after being activated by BCG HSP 70, the DCs pulsed with antigens could keep their special markers and the expression levels of CD54, CD83, CD86 molecules increased too. The stimulatory abilities in allogeneic MLR and proliferation of self lymphocytes also improved. CONCLUSION: This study shows that BCG HSP 70 can induce DCs pulsed with antigens maturation and improve their antigen-presenting ability, which may be a useful maturation inducer for dendritic cells.

In vitro modulation of telomerase activity, telomere length and cell cycle in MKN45 cells by verbascoside.

Screening of natural products with anti-tumor activity as telomerase inhibitor is a new subject in the field of tumor therapy. Using telomerase PCR ELISA, telomere DNA hybridization and flow cytometry analysis, the effects of verbascoside, a phenylpropanoid glucoside extracted from Pedicularis striata Pall, on telomerase activity, telomere length and cell cycle of human gastric carcinoma cells MKN45 was examined in vitro. After being treated with a 50 % inhibition concentration of verbascoside (17.8 microg/ml), telomerase activity in the cells was significantly inhibited but not in the cellular supernatant, the average telomere length became remarkably short, and the sub-G0 /G1 peak and G2/M arrest were also displayed when compared to the control cells. These results suggest that verbascoside mediated-cell differentiation and apoptosis may be affected by telomere-telomerase-cell cycle dependent modulation. Thus, the antitumor mechanism of verbascoside is demonstrated once more by its inhibiting effect on telomerase activity in tumor cells, and the telomerase assay may provide a valuable screening method for antitumor activity of natural products.