Development and thinking of Chinese radiation medicine / 中华疾病控制杂志

In this paper, we reviewed the initiation and development of radiation medicine in China, field researches on the health effects of nuclear test and the great leap from the final reports, advance in clinical diagnosis and treatment of radiation injury, and research of radiation combined injuries. Nowadays, China makes great efforts to move up further in development and peaceful use of nuclear energy as one nuclear power. So, nuclear development and nuclear safety have ushered in new opportunities and challenges. To this end, we must maintain a clear understanding, grasp new opportunities, meet new challenges, and be prepared for danger. Thus, a bright future for research in radiation medicine will come.

Exogenous gene expression in vitro and in vivo in bone marrow mesenchymal stem cells modified by hPDGF-A and hBD(2) / 中国实验血液学杂志

The objective of this study was to investigate the expression of exogenous hPDGF-A and hBD(2) in gene-modified bone marrow mesenchymal stem cells (BM-MSCs) in vitro and in vivo. Recombinant adenovirus vector expressing hPDGF-A/hBD(2) genes was constructed and packaged into virion. Primary isolated and cultured BM-MSCs were transfected by using hPDGF-A hBD(2), then the expressions of exogenous hPDGF-A/hBD(2) were detected by immunocytochemical staining in vitro. The conditioned medium (serum-free cultured supernatant of BM-MSCs transfected with recombinant adenovirus) collected from gene-modified BM-MSCs was applied to scratch wound on monolayer cells of multipotential cell line 10T1/2 in order to confirm the stimulative effect of hPDGF-A on cell migration. Gene-modified BM-MSCs were topically transplanted on wound of rats with radiation and skin excision combined injury. The distribution of BM-MSCs and expression of hPDGF-A/hBD(2) on the wound was observed by fluorescent microscopy and immunohistochemical staining respectively. The results indicated that the rat BM-MSCs transfected with recombinant adenovirus could express the EGFP in vitro. The immunofluorescent cytochemistry assay showed that the gene-modified BM-MSCs expressed the hPDGF-A and hBD(2). The scratch test confirmed that the percentage of healing area of wound in cultured supernatant group of gene-modified BM-MSCs was significant higher than that in control group on 8, 12, 24 and 48 hours (p < 0.05). The fluorescence microscopy of exogenous gene-modified BM-MSCs transplanted on wound revealed that the gene-modified BM-MSCs could higher express exogenous genes of EGFP at least within 2 weeks. The immunohistochemistry staining of wound indicated that the expression of exogenous genes began from day 3, reached to peak on day 7, and still visible on day 21 even though the expression became weak because of the possible dilution of the exogenous genes during cell division. It is concluded that efficient expression of exogenous hPDGF-A/hBD(2) in gene-modified BM-MSCs are demonstrated both in vitro and in vivo, which suggests that the molecular mechanism underlying chronic wound-healing accelerated by the strategy combining cell therapy with gene therapy.

Studies on the treatment of combined radiation-burn injury / 中华烧伤杂志

Combined radiation-burn injuries mainly occur under the circumstances of nuclear explosion, nuclear accident, nuclear terrorism, depleted uranium attack, as well as secondary injuries following attack on nuclear installation. Combination of burn and radiation injuries bring along more serious whole body damage, more complicated pathological mechanism and much more difficult management. Research progress on the pathological mechanism and medical management of several key links of combined injury were discussed in this paper. (1) Enhancement of early first aid and prevention of early death of wounded. (2) Damage and restoration of hemopoietic function. (3) Disturbance of immune function and prevention and treatment of infection (mainly on the intestinal mucosa immunity and enterological infection). (4) Management of burn wound. (5) The role of several important measures in the comprehensive treatment.

Influence of macrophages on the expression of vascular endothelial growth factor receptor mRNA, homeobox B2 mRNA, and integrin alpha nu beta3 in vascular endothelial strain / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of macrophages on the expression of the vascular endothelial growth factor (VEGF) receptor (KDR) mRNA, homeobox B2 (HOXB2) mRNA, and integrin alpha nu beta3 in vitro in vascular endothelial strain.</p><p><b>METHODS</b>Human umbilical vein cells (ECV304) were cultured in vitro and divided into 4 groups, i.e. (1) ECV304 group, (2) ECV304 + conA group [with conA (25 microg/ml in culture) added to ECV304], (3) ECV304 + U937 group (with 1 x 10(5)/ml of U937 cells added to 1 x 10(5)/ml ECV 304), (4) ECV304 + U937 + conA group [with 1 x 10(5)/ml of U937 cells and conA (25 microg/ml in culture)] groups. Forty-eight hours after culturing, the expression of integrin receptor alpha nu beta3 and the changes in the expression of KDR mRNA and HOXB2 mRNA in each group were determined by immunofluorescent technique and RT-PCR, respectively.</p><p><b>RESULTS</b>The expression of integrin receptor alpha nu beta3, KDR mRNA, and HOXB2 mRNA in ECV304 group were 6.7 +/- 1.5, 0.633 +/- 0.012, and 0.674 +/- 0.004, respectively, while those in ECV304 + U937 + conA group (10.2 +/- 1.7, 0.879 +/- 0.003, 0.947 +/- 0.003) were obviously more upregulated when compared with those in ECV304 group (P < 0.01). No difference in the above indices was found between ECV304 and ECV304 + conA, ECV304 + U937 groups (P > 0.05).</p><p><b>CONCLUSION</b>Macrophages activated by ConA can accelerate the proliferation, migration and adhesion to the basement membrane matrix of vascular endothelial cells through the influence on the expression of KDR mRNA, HOXB2 mRNA and integrin alpha nu beta3, and through this pathway the angiogenesis is modulated.</p>

Effect of lipopolysacharide on the biological features and growth factor secretion power of U937 cell line / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effect of lipopolysacharide (LPS) in different concentrations on the biological features and growth factor secretion power of U937 cell line.</p><p><b>METHODS</b>In vitro cultured U937 cells were stimulated by 0 (as control), 0.1, 1.0, 10.0, 50.0 and 100 micro g/ml LPS respectively for 24 hours. Thereafter, the cell proliferation ability was determined by MTT method. The cell apoptosis rate was determined by flow cytometry. The changes in the contents of transforming growth factor beta(1) (TGFbeta(1)) and vascular endothelial growth factor (VEGF) of the supernatant of the cell culture were assessed by ELISA.</p><p><b>RESULTS</b>Apoptosis and TGFbeta(1) secretion could be induced by LPS in dose of 0.1 to 100 micro g/ml when compared with that without LPS challenge (P < 0.05 - 0.01). In detail, LPS in lower dose (0.1, 1.0 and 10.0 micro g/ml) could promote the proliferation of U937 (P < 0.05 - 0.01) but exerted no effect on VEGF secretion. In contrary, LPS in high dose (50 and 100 micro g/ml) could promote VEGF secretion (P < 0.01) but exerted no effects on the proliferation of U937 cells.</p><p><b>CONCLUSION</b>U937 cells could be activated to increase the secretion of TGFbeta(1) by LPS in optimal dose of 0.1 - 10.0 micro g/ml, but the secretion of VEGF could only be promoted by LPS in higher concentration.</p>

The role of p38 on the differentiation of MSCs to myoblasts / 中国应用生理学杂志

<p><b>AIM</b>Inducing mesenchymal stem cells (MSCs) differentiate to myoblasts with 5-azacytidine(5-Aza-CR), investigating the expression of Myf5 and the role of the signal transduction case of p38 in all the course of differentiation.</p><p><b>METHODS</b>Separating and purifying bone marrow-derived MSCs, inducing MSCs differentiation to myoblasts with 10 micromol/L 5-Aza-CR, assaying the gene expression time of Myf5 with RT-PCR method, the antigen expression of myosin with immunohistochemistry method and observing the changes of the activity of phosphorylation p38 before and after inhibited by SB203580 with Western-blot method.</p><p><b>RESULTS</b>MSCs begin to express Myf5 delayed to the 9th day after inhibited by SB203580. Some of MSCs express myosin at the 7th day after induced; The phosphorylation p38 activity of MSCs enhanced after induced by 5-Aza-CR but obviously decreased after inhibited by SB203580.</p><p><b>CONCLUSION</b>MSCs can express myogenic regulator factors and orientation differentiate to myoblasts after induced by 5-Aza-CR, p38 really have a positive signal transduction affection in this course.</p>

The role of homeobox B2 gene in vascular endothelial proliferation and the protective effects of VEGF on the endothelia against radiation injury / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the role of HOXB2 gene in the proliferation of primary human umbilical vein endothelial cells (HUVECs) and the protective effects of VEGF on the endothelia against radiation injury.</p><p><b>METHODS</b>HUVECs were isolated, cultured, subcultured and identified. (1) Liposome coated oligodeoxynucleotide (odn) and homeoboxB2 antisense oligodeoxyncleotide (HOXB2asodn) were prepared prepared in the concentrations of 0.25, 0.5, 1.0 and 2.5 mg/L for the stimulation of HUVEC. (3)H-TdR incorporation test and MTT method were employed to determine the proliferation activity of HUVECs after activation. The cell cycle analysis of HUVECs was determined by flow cytometry. The expression level of HOXB2mRNA within HUVECs was detected by RT-PCR (reverse transcription polymerase chain reaction). (2) HUVECs were separately treated with the addition of VEGF in concentration of 50 microg/L, by radiation in the dose of 6 Gy or 12 Gy (60)Co gamma gamma ray, or radiation with 12 Gy (60)Co gamma gamma ray followed by the addition of VEGF in dose of 50 microg/L. The cellular morphology was observed and the cellular proliferation activity was determined by MTT method.</p><p><b>RESULTS</b>(1) The proliferation activity of HUVECs could be markedly inhibited by liposome coated HOXB2asodn in comparison to liposome-odn (P < 0.05 or 0.001), and the inhibition effect was positively correlated with the increase in asodn concentration. The cell ratio in S phase and the expression level of the HOXB2mRNA could be lowered by asodn in dose of 2.5 mg/L (P < 0.05 or 0.001). (2) Radiation by (60)Co gamma ray could lead to the nuclear enlargement, vacuolation in the cytoplasm, multiplicity of nucleus and nuclear swelling. The proliferative activity of HUVECs was increased from 0.365 +/- 0.047 and 0.487 +/- 0.022 without radiation to 0.557 +/- 0.042 and 0.648 +/- 0.021 24 and 48 hours after 6 Gy radiation However it was decreased to 0.263 +/- 0.038 and 0.306 +/- 0.024 (P < 0.01) after 12 Gy (60)Co gamma ray radiation. Nevertheless, the cell morphology was obviously improved and the proliferation was enhanced by the addition of VEGF after 12 Gy radiation.</p><p><b>CONCLUSION</b>HOXB2 gene played important roles in the biological activities of HUVECs. Small dose (6 Gy) gamma-radiation could promote, but large dose (12 Gy) could decrease the mRNA expression of HOXB2 gene in HUVECs. In addition, VEGF could protect HUVECs against radiation injury.</p>

Effects of total body irradiation injury on the participation of dermal fibroblasts in tissue repair / 生理学报

Wound combined with total body irradiation (TBI) injury results in impairment of tissue repair and delayed processes of healing, so it has been considered as an important and representative model of impaired wound healing, but the mechanism is not fully clarified. Fibroblasts in wound are the most important cells participating in tissue repair, whereas its radiosensitivity is not high. To understand whether TBI injury has direct damaging effects on fibroblasts in wound, fibroblasts in wound combined with TBI injury and in wound of simple incision injury were isolated and cultured, and parameters associated with tissue repair were determined. The results showed that the abilities of proliferation, attachment and adhesion of fibroblasts isolated from wounds combined with TBI injury significantly decreased as compared with those of simple incision injury, nevertheless, apoptotic ratio of fibroblasts isolated from wounds combined with TBI injury increased significantly. These data suggest that TBI injury may cause direct damaging effects on fibroblasts in wounds, which might be one of the dominant reasons for impairment of wound healing when it is combined with TBI injury.

Cultivation and isolation of the bone marrow mesenchymal stem cells / 第三军医大学学报

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

Changes of glucocorticoid receptor in hepatic cytoplasm in scalded rats and its regulation by α-MSH and KPV peptide / 第三军医大学学报

Objective　To observe the changes of glucocorticoi d receptor (GR) in hepatic cytoplasm in rats after scalding-induced pathologic al stress and its regulation. Methods　The receptor binding capa city (R0) and the apparent dissociation constant (Kd) of GR in hepatic cytopla sm of normal, low-degree and heavy-degree scalded rats were measured with rad io-ligand binding assay, with ［3H］ dexamethasone as ligand. The changes of R0 and Kd of GR were regulated by injections of anti-rat TNFα, IL-1β a ntibodies, α-melanocyte-stimulating hormone (α-MSH), and KPV peptide( Ac- D-Lys-L-Pro-D-Val) respectively in vivo. Results　The R 0 of GR in hepatic cytoplasm in rats 12 h after heavy-degree scalding ［Mass action robust: （205.52±30.14） fmol/mg; Scatchard: （208.45±30.78） fmol/mg ］were significantly lower than that of control group ［Mass action robust:（307 .86±24.22） fmol/mg;Scatchard:（306.71±27.96） fmol/mg］(P<0.01), but no s ignificant difference was found in the R0 of GR between the control and the ra ts 12 h after low-degree scalding ［Mass action robust: （285.19±16.62） fmol/ mg ; Scatchard: （296.64±16.06） fmol/mg］. The injection of anti-rat TNFα, IL-1β antibodies, α-MSH and KVP all prevented the decline of R0 of GR in h epatic cytoplasm in rats with severe scalding. Conclusion　The injections of anti-rat TNFα, IL-1β antibodies, α-MSH or KPV can attenuate the reduction of GR in rat hepatic cytoplasm caused by severe scalding-induced pathological stress to some extent.

Effect of glucagon-like peptide 2 on mitogen-activated protein kinase activity in small intestinal epithelia of mice after radiation injury / 第三军医大学学报

Objective　To study the effect of glucagon-like peptide 2 (GLP-2) on mitogen-activated protein kinase (MAPK) activity in small intestinal epithelia in mice after radiation injury and its relation with the change of small intestinal epithelial proliferation. Methods　Mice were given a single dose of 8 Gy of total body 60Co gamma irradiation and then divided into GLP-2 and control groups. The activity of MAPK and proliferation rate in small intestinal epithelia were measured. Results　The activity of MAPK in small intestinal epithelia was higher in GLP-2-treated mice than in irradiated mice, and the proliferation rate in small intestinal epithelia significantly increased in GLP-2-treated mice. These two indices were of significantly positive correlated. Conclusion　GLP-2 can promote small intestinal epithelial proliferation in irradiated mice, and this may be related to activation of MAPK in small intestinal epithelia.

Effects of glucagon-like peptide 2 on recovery of small intestinal epithelia from radiation injury in mice / 第三军医大学学报

Objective　To investigate the effects of glucagon-like peptide 2 (GLP-2) on recovery of small intestinal epithelia from radiation injury in mice. Methods　Mice received a single 8 Gy dose of total body irradiation from 60Co gamma ray followed by treatment with GLP-2 or vehicle. DNA and protein content in small intestinal mucosa were measured, and small intestine was processed for histological examination with light microscope and scanning electron microscope. Results　Small intestinal mucosal DNA and protein content, villus height, and villus number significantly decreased in irradiated mice, partial villus tips were ulcerated. GLP-2 administration caused increase in DNA and protein content, villus height, and villus number as compared with irradiated control group. Meanwhile, the villus tips were lack of ulceration. Conclusion　GLP-2 can promote recovery of small intestinal epithelia from radiation injury in mice.

Effects of irradiation and W11-a12 on anion-selective channel of mouse peritoneal macrophage / 第三军医大学学报

Objective　To study the effects of irradiation and W11-a12，a kind of repair-promoting drug，on anion-selective channel in membranes of mouse peritoneal macrophage. Methods　The activity of anion-selective channel was recorded from cell-attached patches with patch clamp techniques． Results　The effects of irradiation on anion-selective channel in membranes of peritoneal macrophage included：①decreasing the mean number of activated channels by the presence of zymosan; ②prolonging the mean time from stimulus to the opening of channels; ③depressing the opening of channels by decreasing open-state probability，shortening open-time and prolonging close-time． The effects of irradiation could partly be depressed by W11-a12. Conclusion　Irradiation will depress the anion-selective channel of peritoneal macrophage, which may be an important way to depress the function of macrophage．

Effects of systemic irradiation and W11-a12 on neutrophils in wounds / 第三军医大学学报

Objective　To study the effects of systemic irradiation and conglutinant drug W11-a12 on the number and some functions of wound nentrophils (Neu). Methods　Wound Neu was collected from sponges which were implanted in rat's dorsum incision. The number of Neu, as well as the phagocytic function and motility of wound Neu were measured. Results　After 4,6,8 Gy systemic irradiation, the number of white blood cells and Neu in wound, as well as the phagocytic function and chemotactic motility of wound Neu, were significantly decreased at 24 h, 48 h after wounding. W11-a12 markedly increased the number of wound Neu, improved the phagocytic function and chemotactic motility of wound Neu at 24 h, 48 h after wounding despite the rats were radiated or not. Conclusion　The results indicated that the decreased number and function of wound Neu in the early stage of wound healing contributed to the impairment of repair after systemic irradiation. W11-a12 accelerated normal and irradiation-impaired wound healing partly by increasing the number of wound Neu and improving the Neu function.

Cultivation and isolation of the bone marrow mesenchymal stem cells / 第三军医大学学报

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

Changes of glucocorticoid receptor in hepatic cytoplasm in scalded rats and its regulation by α-MSH and KPV peptide / 第三军医大学学报

Objective　To observe the changes of glucocorticoi d receptor (GR) in hepatic cytoplasm in rats after scalding-induced pathologic al stress and its regulation. Methods　The receptor binding capa city (R0) and the apparent dissociation constant (Kd) of GR in hepatic cytopla sm of normal, low-degree and heavy-degree scalded rats were measured with rad io-ligand binding assay, with ［3H］ dexamethasone as ligand. The changes of R0 and Kd of GR were regulated by injections of anti-rat TNFα, IL-1β a ntibodies, α-melanocyte-stimulating hormone (α-MSH), and KPV peptide( Ac- D-Lys-L-Pro-D-Val) respectively in vivo. Results　The R 0 of GR in hepatic cytoplasm in rats 12 h after heavy-degree scalding ［Mass action robust: （205.52±30.14） fmol/mg; Scatchard: （208.45±30.78） fmol/mg ］were significantly lower than that of control group ［Mass action robust:（307 .86±24.22） fmol/mg;Scatchard:（306.71±27.96） fmol/mg］(P<0.01), but no s ignificant difference was found in the R0 of GR between the control and the ra ts 12 h after low-degree scalding ［Mass action robust: （285.19±16.62） fmol/ mg ; Scatchard: （296.64±16.06） fmol/mg］. The injection of anti-rat TNFα, IL-1β antibodies, α-MSH and KVP all prevented the decline of R0 of GR in h epatic cytoplasm in rats with severe scalding. Conclusion　The injections of anti-rat TNFα, IL-1β antibodies, α-MSH or KPV can attenuate the reduction of GR in rat hepatic cytoplasm caused by severe scalding-induced pathological stress to some extent.

Effect of glucagon-like peptide 2 on mitogen-activated protein kinase activity in small intestinal epithelia of mice after radiation injury / 第三军医大学学报

Objective　To study the effect of glucagon-like peptide 2 (GLP-2) on mitogen-activated protein kinase (MAPK) activity in small intestinal epithelia in mice after radiation injury and its relation with the change of small intestinal epithelial proliferation. Methods　Mice were given a single dose of 8 Gy of total body 60Co gamma irradiation and then divided into GLP-2 and control groups. The activity of MAPK and proliferation rate in small intestinal epithelia were measured. Results　The activity of MAPK in small intestinal epithelia was higher in GLP-2-treated mice than in irradiated mice, and the proliferation rate in small intestinal epithelia significantly increased in GLP-2-treated mice. These two indices were of significantly positive correlated. Conclusion　GLP-2 can promote small intestinal epithelial proliferation in irradiated mice, and this may be related to activation of MAPK in small intestinal epithelia.

Effects of glucagon-like peptide 2 on recovery of small intestinal epithelia from radiation injury in mice / 第三军医大学学报

Objective　To investigate the effects of glucagon-like peptide 2 (GLP-2) on recovery of small intestinal epithelia from radiation injury in mice. Methods　Mice received a single 8 Gy dose of total body irradiation from 60Co gamma ray followed by treatment with GLP-2 or vehicle. DNA and protein content in small intestinal mucosa were measured, and small intestine was processed for histological examination with light microscope and scanning electron microscope. Results　Small intestinal mucosal DNA and protein content, villus height, and villus number significantly decreased in irradiated mice, partial villus tips were ulcerated. GLP-2 administration caused increase in DNA and protein content, villus height, and villus number as compared with irradiated control group. Meanwhile, the villus tips were lack of ulceration. Conclusion　GLP-2 can promote recovery of small intestinal epithelia from radiation injury in mice.

Effects of irradiation and W11-a12 on anion-selective channel of mouse peritoneal macrophage / 第三军医大学学报

Objective　To study the effects of irradiation and W11-a12，a kind of repair-promoting drug，on anion-selective channel in membranes of mouse peritoneal macrophage. Methods　The activity of anion-selective channel was recorded from cell-attached patches with patch clamp techniques． Results　The effects of irradiation on anion-selective channel in membranes of peritoneal macrophage included：①decreasing the mean number of activated channels by the presence of zymosan; ②prolonging the mean time from stimulus to the opening of channels; ③depressing the opening of channels by decreasing open-state probability，shortening open-time and prolonging close-time． The effects of irradiation could partly be depressed by W11-a12. Conclusion　Irradiation will depress the anion-selective channel of peritoneal macrophage, which may be an important way to depress the function of macrophage．

Effects of systemic irradiation and W11-a12 on neutrophils in wounds / 第三军医大学学报

Objective　To study the effects of systemic irradiation and conglutinant drug W11-a12 on the number and some functions of wound nentrophils (Neu). Methods　Wound Neu was collected from sponges which were implanted in rat's dorsum incision. The number of Neu, as well as the phagocytic function and motility of wound Neu were measured. Results　After 4,6,8 Gy systemic irradiation, the number of white blood cells and Neu in wound, as well as the phagocytic function and chemotactic motility of wound Neu, were significantly decreased at 24 h, 48 h after wounding. W11-a12 markedly increased the number of wound Neu, improved the phagocytic function and chemotactic motility of wound Neu at 24 h, 48 h after wounding despite the rats were radiated or not. Conclusion　The results indicated that the decreased number and function of wound Neu in the early stage of wound healing contributed to the impairment of repair after systemic irradiation. W11-a12 accelerated normal and irradiation-impaired wound healing partly by increasing the number of wound Neu and improving the Neu function.