The status and model of children primary nephrotic syndrome in continuing nursing.

BACKGROUND: Nephrotic syndrome (NS) is a common glomerular disease in children. Nursing during hospitalization alone cannot solve the psychological, physiological and social health problems of children. Continuing care models may provide patients with more continuous and efficient care services. Therefore, the present study aimed to provide theoretical support for the implementation and development of children's primary nephrotic syndrome (PNS) and children's chronic disease continuing nursing through the construction of a children's PNS continuing nursing model. METHODS: Each item of the transitional care model for children with PNS was demonstrated using the Delphi method for two rounds of correspondence. The main items included four components: the composition of personnel, the responsibilities of each member, the content of work, and the evaluation indicators. RESULTS: A transitional care model for children with PNS was formed. The expert judgment coefficient of two rounds of correspondence was 0.84, the familiarity degree coefficient was 0.76, the authority degree coefficient was 0.80, the coefficient of variation was between 0.02 and 0.23, and the coordination coefficient was 0.458 and 0.327, respectively (P<0.01). CONCLUSIONS: The experts in the present research were highly motivated, had a high degree of authority, and presented consistent opinions. Hence, the construction of a transitional care model for children with PNS is scientifically feasible.

Extraction of essential oil from Citrus reticulate Blanco peel and its antibacterial activity against Cutibacterium acnes (formerly Propionibacterium acnes).

Citrus is one of the largest output fruits in the word. In China, the major orange variety is the Citrus reticulate Blanco (Ponkan). The peels are discarded as waste material, its comprehensive utilization is urgently needed. In this work, hydrodistillation method was developed to extract citrus essential oil (EO) from Blanco peel. With the optimal extraction conditions, the EO yield was more than 3%. By GC-MS analysis, 53 compounds were identified from the citrus EO. Terpenes compounds accounted for 71.2%, especially d-limonene (major composition) accounted for 58.9%. The obtained citrus EO showed remarkable antibacterial activity against Cutibacterium acnes (C. acnes, Formerly P. acnes) and common microorganisms such as S. aureus, B. subtilis, and E. coli. Even compared with the common antibiotics (such as erythromycin, clindamycin, and tetracycline) for acne therapy, its antibacterial activity against C. acnes is more excellent. This work provides a potential therapy material for the treatment of acne.

Characterization of different biodegradable scaffolds in tissue engineering.

The aim of the present study was to compare the characteristics of acellular dermal matrix (ADM), small intestinal submucosa (SIS) and Bio­Gide scaffolds with acellular vascular matrix (ACVM)­0.25% human­like collagen I (HLC­I) scaffold in tissue engineering blood vessels. The ACVM­0.25% HLC­I scaffold was prepared and compared with ADM, SIS and Bio­Gide scaffolds via hematoxylin and eosin (H&E) staining, Masson staining and scanning electron microscope (SEM) observations. Primary human gingival fibroblasts (HGFs) were cultured and identified. Then, the experiment was established via the seeding of HGFs on different scaffolds for 1, 4 and 7 days. The compatibility of four different scaffolds with HGFs was evaluated by H&E staining, SEM observation and Cell Counting Kit­8 assay. Then, a series of experiments were conducted to evaluate water absorption capacities, mechanical abilities, the ultra­microstructure and the cytotoxicity of the four scaffolds. The ACVM­0.25% HLC­I scaffold was revealed to exhibit the best cell proliferation and good cell architecture. ADM and Bio­Gide scaffolds exhibited good mechanical stability but cell proliferation was reduced when compared with the ACVM­0.25% HLC­I scaffold. In addition, SIS scaffolds exhibited the worst cell proliferation. The ACVM­0.25% HLC­I scaffold exhibited the best water absorption, followed by the SIS and Bio­Gide scaffolds, and then the ADM scaffold. In conclusion, the ACVM­0.25% HLC­I scaffold has good mechanical properties as a tissue engineering scaffold and the present results suggest that it has better biological characterization when compared with other scaffold types.

A Novel Cellulase Produced by a Newly Isolated Trichoderma virens.

Screening and obtaining a novel high activity cellulase and its producing microbe strain is the most important and essential way to improve the utilization of crop straw. In this paper, we devoted our efforts to isolating a novel microbe strain which could produce high activity cellulase. A novel strain Trichoderma virens ZY-01 was isolated from a cropland where straw is rich and decomposed, by using the soil dilution plate method with cellulose and Congo red. The strain has been licensed with a patent numbered ZL 201210295819.6. The cellulase activity in the cultivation broth could reach up to 7.4 IU/mL at a non-optimized fermentation condition with the newly isolated T. virens ZY-01. The cellulase was separated and purified from the T. virens culture broth through (NH4)2SO4 fractional precipitation, anion-exchange chromatography and gel filtration chromatography. With the separation process, the CMC specific activity increased from 0.88 IU/mg to 31.5 IU/mg with 35.8 purification fold and 47.04% yield. Furthermore, the enzymatic properties of the cellulase were investigated. The optimum temperature and pH is 50 °C and pH 5.0 and it has good thermal stability. Zn2+, Ca2+ and Mn2+ could remarkably promote the enzyme activity. Conversely, Cu2+ and Co2+ could inhibit the enzymatic activity. This work provides a new highly efficient T. virens strain for cellulase production and shows good prospects in practical application.

Exogenous normal lymph alleviates lipopolysaccharide-induced acute kidney injury in rats.

BACKGROUND: Acute kidney injury (AKI) is a common pathological process which occurs in hemorrhage, intoxication, etc. It has been shown that the lymphatic circulation plays an important regulatory role in the pathogenesis of hemorrhage shock, and that exogenous normal lymph (ENL) has a beneficial effect on multiple organ injuries. In the present study, we investigated the effect of ENL on lipopolysaccharide (LPS)-induced AKI in rats. METHODS: The AKI was induced by the jugular vein injection of LPS (iv, 15 mg/kg). After 15 min of LPS injection, saline or ENL without cell components (5 mL/kg) was iv infused at the speed of 0.5 mL per minute. Then, the renal function indices in plasma and renal histomorphology, and the levels of P-selectin, intercellular adhesion molecule-1 (ICAM-1), myeloperoxidase (MPO) and Na(+)-K(+)-ATPase in renal tissue were assessed at 3 or 6 h after LPS injection. RESULTS: LPS induced a severe kidney injury including increased levels of urea, creatinine in plasma, aggrandized activities of ICAM-1 and MPO in renal tissue, and decreased the Na(+)-K(+)-ATPase activity in renal cells. These deleterious effects of LPS were significantly ameliorated by ENL treatment. CONCLUSION: The present results indicate that ENL protect against LPS-induced AKI, suggesting an alternative therapeutic strategy for treatment of kidney injury accompanied with severe infection or sepsis.

Exogenous normal lymph alleviates microcirculation disturbances and abnormal hemorheological properties in rats with disseminated intravascular coagulation.

Disturbances of the microcirculation and abnormal hemorheological properties are important factors that play an important role in disseminated intravascular coagulation (DIC) and result in organ dysfunction or failure. In the present study, we established an animal model of DIC using intravenous Dextran 500 in rats, and used exogenous normal lymph corresponding to 1/15 of whole blood volume for injection through the left jugular vein. We found that normal lymph could improve the blood pressure and survival time of rats with DIC. The results regarding the mesenteric microcirculation showed that the abnormality of the diameter of mesenteric microvessels and micro-blood flow speed in the DIC+lymph group was significantly less than in the DIC+saline group. Whole blood viscosity, relative viscosity, plasma viscosity, hematocrit (Hct), erythrocyte sedimentation rate (ESR), and electrophoresis time of erythrocytes were significantly increased in the DIC+saline group compared to the control group. The electrophoretic length and migration of erythrocytes from the DIC+saline and DIC+lymph groups were significantly slower than the control group. Blood relative viscosity, Hct, ESR, and electrophoretic time of erythrocytes were significantly increased in the DIC+lymph group compared to the control group. Whole blood viscosity, relative viscosity and reduced viscosity were significantly lower in the DIC+lymph group than in the DIC+saline group, and erythrocyte deformability index was also significantly higher than in the DIC+saline and control groups. These results suggest that exogenous normal lymph could markedly improve the acute microcirculation disturbance and the abnormal hemorheological properties in rats with DIC induced by Dextran 500.

Exogenous normal lymph alleviates microcirculation disturbances and abnormal hemorheological properties in rats with disseminated intravascular coagulation

Disturbances of the microcirculation and abnormal hemorheological properties are important factors that play an important role in disseminated intravascular coagulation (DIC) and result in organ dysfunction or failure. In the present study, we established an animal model of DIC using intravenous Dextran 500 in rats, and used exogenous normal lymph corresponding to 1/15 of whole blood volume for injection through the left jugular vein. We found that normal lymph could improve the blood pressure and survival time of rats with DIC. The results regarding the mesenteric microcirculation showed that the abnormality of the diameter of mesenteric microvessels and micro-blood flow speed in the DIC+lymph group was significantly less than in the DIC+saline group. Whole blood viscosity, relative viscosity, plasma viscosity, hematocrit (Hct), erythrocyte sedimentation rate (ESR), and electrophoresis time of erythrocytes were significantly increased in the DIC+saline group compared to the control group. The electrophoretic length and migration of erythrocytes from the DIC+saline and DIC+lymph groups were significantly slower than the control group. Blood relative viscosity, Hct, ESR, and electrophoretic time of erythrocytes were significantly increased in the DIC+lymph group compared to the control group. Whole blood viscosity, relative viscosity and reduced viscosity were significantly lower in the DIC+lymph group than in the DIC+saline group, and erythrocyte deformability index was also significantly higher than in the DIC+saline and control groups. These results suggest that exogenous normal lymph could markedly improve the acute microcirculation disturbance and the abnormal hemorheological properties in rats with DIC induced by Dextran 500.

Exogenous normal lymph alleviating kidney injury by improving coagulation function in disseminated intravascular coagulation rats.

AIM: To investigate the effect of exogenous normal lymph on kidney injury in disseminated intravascular coagulation (DIC) rats and to probe its mechanism. METHODS: The DIC model was established by intravenous injection of Dextran 500. After 6 min, normal lymph without cell components was infused in the lymph group. After 40 min, the renal and coagulation function indices and renal histomorphology were observed. RESULTS: Serum urea and creatinine in the model group were significantly higher than in the control and lymph groups. Renal morphological study showed red blood cell silting and casts forming in the model group. The prothrombin time (PT), prothrombin time ratio (PTR), activated partial thromboplastin time (APTT), and thrombin time of lymph and model groups were higher than those in the control group, whereas fibrinogen was lower. The PT, PTR, and APTT were prolonged in the lymph group than in the model group. The platelet functions of the lymph and model groups were higher than in the control group, but platelet aggregation rate and thrombosis-forming indices were lower than in the control group; the platelet adhesive and aggregation rates and thrombosis dry weight of the lymph group were lower than those of the model group. CONCLUSION: Exogenous normal lymph could alleviate kidney injury in DIC rats, which may be related to the improving coagulation function.

The mechanism of spleen injury in rabbits with acute renal failure.

Immune function disorders are common during acute renal failure (ARF), but the mechanisms are unknown. As the spleen is the largest organ of the immune system, we aimed to observe if there are morphological changes in the spleen in rabbits with ARF. In addition, we tried to explore its mechanism from the perspective of oxygen free radicals, nitric oxide (NO), myeloperoxidase (MPO), and membrane pump activities. ARF animal models were established by either hypodermic injection of 1.3 mL/kg bw 1% HgCl2 or intramuscular injection of 10 mL/kg bw 50% glycerin. Animals were divided into 12 h, 24 h, and 48 h treatment groups with six rabbits in each group. Compared with control animals, congestion was found in the spleen and splenic trabeculae were increased in the two ARF model groups at multiple time points. The malonaldehyde, NO, nitric oxide synthase, and MPO levels in the ARF models were increased compared with the control group at 24 h or 48 h, and the superoxide dismutase and adenosine triphosphatase activities were significantly lower than the levels in the control group at multiple time points. These indices of free radical damage were induced gradually with ARF development, and there were statistically significant differences at different time points. These data suggested that histological damage of spleen during ARF may lead to immune disorders, which might be related to free radical injury, NO excessive release, polymorphonuclear neutrophils (PMN) sequestration, and membrane pump dysfunction.

[Mechanism of multiple organ injury subsequent to acute renal failure with respect to membrane pump activity in rabbits].

OBJECTIVE: To observe the changes in membrane pump activity of kidney, myocardium and pancreas in rabbits with acute renal failure (ARF) in rabbits, and inquire into the mechanism of multiple organ injury subsequent to ARF. METHODS: Forty-two rabbits were randomly divided into control group, HgCl(2) group and glycerine group, and the latter two groups were subdivided into 12, 24, 48-hour subgroups, with 6 rabbits in each group. The ARF model was reproduced by hypodermic injection 1% HgCl(2) (1.3 ml/kg) in HgCl(2) group, or intramuscularly injection 50% glycerine (10 ml/kg) in glycerine group, respectively. At different time points, the kidney, myocardium and pancreas were harvested, and homogenates of them were prepared. The ATPase activities of different organ homogenates were determined. RESULTS: It showed that the activities of Na(+)-K(+)-ATPase, Ca(2+)-ATPase, Ca(2+)-Mg(2+)-ATPase of renal homogenate in two model groups were reduced gradually with worsening of renal function, and they became lowest at 48 hours [(0.84±0.16), (0.52±0.17), (0.45±0.09) µmol×mg(-1)×h(-1) in HgCl(2) group; (0.85±0.22), (0.49±0.21), (0.54±0.17) µmol×mg(-1) ×h(-1)in glycerine group]. The respective activities of Na (+)-K (+)-ATPase, Ca (2+)-ATPase, Mg(2+)-ATPase, Ca(2+)-Mg(2+)-ATPase of myocardium and pancreas homogenates in two model groups were reduced gradually following depression of renal function, and they became lowest at 48 hours [(0.56±0.11), (0.51±0.19), (0.55±0.19), (0.37±0.19) µmol×mg(-1)×h(-1) in HgCl(2) group and (0.52±0.19), (0.62±0.10), (0.61±0.16), (0.54±0.10) µmol×mg(-1) ×h(-1) in myocardium homogenate of glycerine group; (0.81±0.12), (0.71±0.15), (0.73±0.18), (0.62±0.16) µmol×mg(-1) ×h(-1) in HgCl(2) group and (0.72±0.13), (0.57±0.18), (0.66±0.14), (0.59±0.23) µmol×mg(-1) ×h(-1) in pancreas homogenate of glycerine group], there was statistical difference compared with control group (P<0.05 or P<0.01). CONCLUSION: The mechanism of myocardial and pancreatic injury subsequent to ARF might be related to reduction of the activity of cell membrane pump.

The mechanism of myocardium and pancreas injury in rabbits with acute renal failure might be related to myeloperoxidase and membrane pump activities.

There is increasing evidence indicating that the distant organ injury is a major contributor of high mortality in patients subjected to acute renal failure (ARF). However, sources and mechanisms that ARF causes distant organ injury remain to be determined. The aim of this study is to explore the mechanism from polymorphonuclear neutrophil (PMN) sequestration and membrane pump suppression. To achieve this, we examined myeloperoxidase (MPO), a marker of PMN accumulation in tissues, and membrane pump activities of heart, pancreas, and kidney in two ARF rabbit models. Rabbits are randomly assigned to control, HgCl(2)-treated, and glycerin-treated groups. ARF animal models are established by hypodermic injection of 1% HgCl(2) with 1.3 mL/kg bodyweight (bw) in HgCl(2)-treated group or intramuscular injection of 50% glycerin with 10 mL/kg bw in glycerin-treated group, respectively, and all animals in each group are further divided into 12 h, 24 h, and 48 h subgroups with each consisting of six rabbits. Six healthy rabbits serve as control group. Results have shown that MPO activities of kidney, myocardium, and pancreas in two model groups were significantly increased than control group at diverse time points. Membrane pump activities of kidney in two model groups are significantly lower than the control group at multiple time points. Moreover, Na(+)-K(+)-, Ca(2+)-, Mg(2+)-, and Ca(2+)-Mg(2+)-ATPase activities of myocardium and pancreas in two model groups are gradually declined with the development of ARF. These findings suggest that PMN sequestration and membrane pump suppression plays an important role in the pathogenesis of ARF and also a major mechanism of myocardium and pancreas injury during the process of ARF.

Mesenteric lymph duct ligation against renal injury in rats after hemorrhagic shock.

BACKGROUND: The kidney is a common target in multiple organ dysfunction syndrome (MODS). The aim of this study is to determine the role of intestinal lymphatic pathway on renal injury in hemorrhagic shock rats. METHODS: Wistar rats were divided into sham, shock, and ligation groups. The hemorrhagic shock model was induced in the shock and ligation groups. After resuscitation, the mesenteric lymph ducts were ligated in the ligation group. Blood from the carotid artery was taken to determine renal functional indices. The kidneys were used to observe histomorphological changes at 6 h after resuscitation. In addition, kidney homogenate was used to determine malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor alpha (TNFalpha), interleukin-6 (IL-6), and myeloperoxidase (MPO) levels at 90 min after shock and 0, 1, 3, 6, 12, and 24 h after resuscitation. And the survival rate of 24 h was recorded. RESULTS: The survival rate in shock group was obviously lower than sham and ligation groups. The urea and creatinine contents in the serum of shock and ligation groups were significantly higher than the sham group; the indices in the ligation group were significantly lower than the shock group. Histological studies showed various degrees of renal injury in the shock and ligation groups with a lesser severity in the ligation group. MDA, TNFalpha, IL-6, and MPO in renal homogenate of the shock group were raised, and the activity of SOD was lower in comparison to the sham group. Further, MDA, TNFalpha, IL-6, and MPO in renal homogenate of the ligation group at 6, 12, and 24 h were lower, and the SOD activity was higher than that of the shock group at the same time points. CONCLUSION: The mesenteric lymph duct ligation could be used to attenuate renal injury in shock rats. Its mechanism might be related to reducing the polymorph nuclear (PMN) and decreasing inflammatory mediator and free radical.

Pancreatic injury in rabbits with acute renal failure.

BACKGROUND: Acute renal failure (ARF) is a common critical disorder. To decrease the mortality, it is important to prevent ARF from invading other organs in clinical setting. It is not known whether there is a dysfunction in pancreas during the pathogenesis of ARF. This study aimed to investigate the changes of morphology and function on pancreas in ARF rabbits. METHODS: Sixty rabbits were randomly divided into four groups. The ARF model of groups 1 and 2 rabbits was established by hypodermic injection of 1% HgCl(2) (1.3 mL/kg) and intramuscular injection of 50% glyceritum (10 mL/kg), respectively. The control groups 1 and 2 were injected with same volume of normal saline. After 24 hours, urea and creatinine contents and pancreatic amylase (AMY) activities in serum were measured using an automatic biochemical analyzer; the insulin levels were measured with radioimmunoassay method. Moreover, morphological alterations were examined by light microscopy; free radicals, nitric oxide (NO), and nitric oxide synthase (NOS) in pancreas homogenate were determined. RESULTS: Morphological study showed that there were vacuolar degeneration and necrosis in pancreas of ARF for both groups 1 and 2. Compared with corresponding control group, the AMY activity was significantly elevated, whereas the INS values were decreased significantly in ARF groups 1 and 2. Malonaldehyde, NO, and NOS in pancreas homogenate were significantly increased, and superoxide dismutase activity was decreased. CONCLUSION: These data suggested that there were morphological damage of pancreas and disturbance of pancreatic secretion function in rabbits with ARF. Free radicals-injury and NO excessive release may explain the observed dysfunction.

[Construction of eukaryotic expression vector of short hairpin RNA targeting human xylosyltransferase-I gene].

OBJECTIVE: To design and construct the plasmids expressing short hairpin RNA (shRNA) targeting human xylosyltransferase- I (XT- I) which is the initiating enzyme in the biosynthesis of proteoglycans (PC). METHODS: Short chain oligonucleotides were designed according to the sequence of XT-I provided by GenBank. The DNA segments were gained through annealing after chemosynthesis, and were cloned into Pgenesil-1 vector. The recombinant XT- I shRNA expression vectors were identified by digestion and sequencing analysis. At last the constructed XT-I expression vectors were transfected into salivary adenoid cystic carcinoma cell line (ACC-M) by Lipofectomine 2000. The expression of green fluorescent protein (GFP) was detected by inverted fluorescent microscope and the rates of transfection were detected by flow cytometer. Semiquantitative RT-PCR was used to detect the expression of mRNA level of XT- I in transfected ACC-M cells and the protein expression of XT- I was detected by Western blot. RESULTS: The plasmids expressing shRNA targeting XT-I gene are called WJ1, WJ2, WJ3, WJ4, WJ5 and WJ6. Successful constructions were identified by digestion and sequencing. The mean rate of transfection was 50.26%. ACC-M cells transfected with WJ1-WJ6 expressed GFP successfully. And by RT-PCR and Western blot, WJ3 showed the most powerful RNAi gene silencing of inhibitory. The inhibition rate was 72.39% of mRNA level and 70.18% of protein level respectively. CONCLUSION: The XT-I shRNA expression vectors were constructed successfully which lays the foundation for RNAi study on the biosynthesis of PG in salivary gland tumors.

[wt-p53 gene repair for p53 mutations of salivary pleomorphic adenoma cells].

OBJECTIVE: To analyze the original mutated codon of p53 gene of salivary pleomorphic adenoma (SPA) and to evaluate the repair effects of wt-p53 on SPA cells. METHODS: Four cases of SPA were obtained from clinical fresh samples and SPA cells were separated and cultured, and then the cells were transduced by Ad-wt-p53. The cells and the corresponding tumor tissue DNA were extracted, PCR and single strand conformational polymorphism (SSCP) and DNA sequencing analysis were performed. RESULTS: PCR-SSCP analysis showed 3 out of 4 SPA with abnormal exon 8 and abnormal exon 6. DNA sequencing analysis showed that exon 6 point mutation was found at codon 203 (GTG-->GCG), poly-codon mutations were found in exon 8 at codon 272 (GTG-->GT square), 275 (TGT-->T square T), 276 (GCC--> square CC) and at codon 290 (CGC-->CGCC). After transduced by Ad-wt-p53, all of the mutated codons were repaired. CONCLUSIONS: p53 gene mutation involved many codons that occurred frequently in the tumorigenesis of SPA. Exogenous wt-p53 could compensate and repair all the mutated p53 codons of SPA cells. SPA cells transduced by Ad-wt-p53 showed the typical apoptosis.

[Role of intestinal lymphatic pathway in pathogenesis of intestine-derived bacteria/endotoxin translocation in rats in shock].

OBJECTIVE: To observe the changes of toxic substances in mesenteric lymph and portal vein blood of rats in hemorrhagic shock, and the influence of mesenteric lymph duct ligation on level of endotoxin (ET) in organs and bacterial contents in mesenteric lymph nodes (MLN) and spleen in rats with hemorrhagic shock, and to evaluate the role of lymphatic pathway in pathogenesis of intestine-derived bacteria/endotoxin translocation (BET) in rats with shock. METHODS: Twenty-four male Wistar rats were randomly divided into the shock group and control group. A model of serious hemorrhagic shock was reproduced by blood shedding to maintain the blood pressure at 40 mm Hg (1 mm Hg=0.133 kPa) for 90 minutes under aseptic condition, and MLN and portal vein blood were harvested. The specimens were also obtained in control group. The contents of ET, tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were determined in them. Thirty male Wistar rats were randomly divided into the sham operation group, shock group and lymphatic duct ligation group. Mesenteric lymph ducts were ligated after resuscitation. All rats were sacrificed, and lung, liver, heart and kidney were removed and homogenized for determination of the content of ET. MLN and spleen homogenates were subjected to bacterial culture. RESULTS: The contents of ET, TNF-alpha and IL-6 in lymph were significantly higher than those of plasma in shock group, and also higher than that in normal plasma and normal lymph (all P<0.01). In shock group the contents of ET in lung, liver, heart and renal homogenate 3 and 6 hours after transfusion and resuscitation were significantly higher than those of sham operation group and ligation group (P<0.05 or P<0.01). Bacterial culture of MLN and spleen in shock group rats 3 and 6 hours after transfusion and resuscitation was positive, but it was not in ligation group. CONCLUSION: The results demonstrate that the intestinal lymphatic pathway plays an important role after compromise of gut barrier function in carrying out BET after hemorrhagic shock.

[The effects of HSV-tk suicide gene and wild-type p53 gene on pleomorphic adenoma cells of salivary gland].

OBJECTIVE: To study the therapeutic effects of combined gene therapy of wild type p53 (wt-p53) and herpes simplex virus thymidine kinase (HSV-tk) gene on pleomorphic adenoma cells of salivary gland. METHODS: Wild type p53 and HSV-tk gene were transfected into human pleomorphic adenoma cells of salivary gland by using recombinant adenovirus vector. The efficiency of transfection was checked and gene was expressed by RT-PCR methods. The cell inhibition after transfected was verified by light microscope and MTT. RESULTS: The proliferation of the pleomorphic adenoma cells transfected wt-p53 and HSV-tk gene was inhibited and the cell survival rate decreased to 54% and 38% respectively in 5 days. However, when wt-p53 gene combined with HSV-tk/GCV system, the killing effects was significantly stronger (P < 0.05) and the cell survival rate decreased to 20%. CONCLUSION: Combining p53 gene with HSV-tk/GCV system for gene therapy in pleomorphic adenoma cells of salivary gland is a valuable method.

[Effects of free oxygen radical and nitric oxide in multiple organ dysfunction syndrome induced by acute renal failure].

OBJECTIVE: To observe the changes in free oxygen radical and nitric oxide (NO) in the liver, lung, and heart homogenate in rabbits with acute renal failure (ARF), and inquire into the mechanism of multiple organ dysfunction syndrome (MODS) subsequent to ARF. METHODS: Sixty rabbits were randomly divided into four groups (n=15 in each group). In the group I ARF was reproduced by hypodermic injection of 1% HgCl(2) in a dose of 1.3 ml/kg, and in group II ARF was reproduced by intramuscular injection of 50% glycerin in a dose of 10 ml/kg. In the control groups I and II the rabbits received an equal volume of normal saline. After 24 hours, blood samples were obtained from all the rabbits, and the liver, lung and heart were harvested and 10% homogenates were made. The blood urea nitrogen and serum creatinine were determined by automatic biochemical analyzer (Aeroset), superoxide dismutase (SOD), malondialdehyde (MDA), NO, nitric oxide synthase (NOS), and inducible nitric oxide synthase (iNOS) in tissue homogenate were also determined. RESULTS: It showed that SOD activity was significantly lowered and the MDA, NO, NOS and iNOS were significantly increased in heart homogenate of ARF groups I and II compared with control group I and II (P<0.01 or P<0.05). But these indexes in the liver and lung homogenates were not significantly different between the groups (all P>0.05). The levels of NO, NOS and iNOS in serum of ARF groups I and II were significantly increased compared with control group I and II (P<0.05 or P<0.01), and the NO contents in renal homogenate of ARF groups I and II were significantly increased compared with control group I and II (P<0.01). CONCLUSION: There is sign of damage of the myocardium in ARF, resulting in MODS. Its possible pathogenic mechanism might be attributable to free oxygen radical injury and increase in NO production. NO plays a dual role of protection and damage in the pathogenesis of ARF.