[Management strategy of femoral artery pseudoaneurysm combined with infectious wounds].

Objective: To investigate the surgical treatment methods of femoral artery pseudoaneurysm combined with infectious wounds and to evaluate the clinical effects. Methods: The retrospective observational research method was used. Twelve patients with femoral artery pseudoaneurysm combined with infectious wounds who met the inclusion criteria were admitted to Nanjing University of Chinese Medicine Wuxi Integrated Traditional Chinese and Western Medicine Hospital (Affiliated Hospital of Jiangnan University) from October 2014 to September 2022, including 6 males and 6 females, aged from 46 to 78 years. In the primary operation, debridement, tumor resection, and artery suture/venous grafting to repair the artery/artery ligation were performed, and the wound area after tumor resection ranged from 4.0 cm×1.5 cm to 12.0 cm×6.5 cm. Wounds that could be sutured were treated with tension reduction suture and extracutaneous continuous vacuum sealing drainage (VSD), while large wounds that could not be sutured were treated with VSD to control infection. In the secondary operation, tension reduction suture was performed to repair the wounds that could be sutured; large wounds were repaired with adjacent translocated flaps with area of 9.0 cm×5.0 cm to 15.0 cm×7.0 cm. Additionally, when the length of the exposed femoral artery was equal to or over 3.0 cm, the wounds were repaired with additional rectus femoris muscle flap with length of 15.0 to 18.0 cm. The donor areas of the flaps were directly sutured. The wound with artery ligation was treated with stamp skin grafting and continuous VSD. The bacterial culture results of the wound exudate samples on admission were recorded. The intraoperative blood loss, the location of femoral artery rupture, the artery treatment method, and the wound repair method in the primary operation were recorded, and the durations of catheter lavage, catheter drainage, and VSD treatment, and the drainage volume after the operation were recorded. The repair method of wounds in the secondary operation, the durations of catheter drainage and VSD treatment, and the total drainage volume after the operation were recorded. The survivals of flap/muscle flap/stamp skin grafts were observed, and the wound healing time was recorded. Follow-up after discharge was performed to evaluate the quality of wound healing and the walking function and to check whether the pulsatile mass disappeared. B-ultrasound or computed tomography angiography (CTA) was performed again to observe potential pseudoaneurysm recurrence and evaluate the patency of blood flow of the femoral artery. Results: The bacterial culture results of wound exudate samples of all the patients were positive on admission. The blood loss was 150 to 750 mL in the primary operation. The arterial ruptures were located in the femoral artery in 8 cases, in the external iliac artery in 2 cases, and in the femoral arteriovenous fistula in 2 cases. Six cases received direct artery suture, 4 cases received autologous great saphenous vein grafting to repair the artery, 1 case received autologous great saphenous vein bypass surgery, and 1 case received artery ligation. The primary wound suture was performed in 4 cases, along with catheter lavage for 3 to 5 days, catheter drainage for 4 to 6 days, VSD treatment for 5 to 7 days, and a total drainage volume of 80 to 450 mL after the surgery. In the secondary operation, the wounds were sutured directly in 3 cases along with catheter drainage for 2 to 3 days, the wound was repaired with scalp stamp skin graft and VSD treatment for 5 days in 1 case, the wounds were repaired with adjacent translocated flaps in 2 cases with catheter drainage for 2 to 3 days, and the wounds were repaired with rectus femoris muscle flaps+adjacent translocated flaps in 2 cases with catheter drainage for 3 to 5 days . The total drainage volume after the secondary operation ranged from 150 to 400 mL. All the skin flaps/muscle flaps/skin grafts survived after operation. The wound healing time ranged from 15 to 36 days after the primary operation. Follow-up of 2 to 8 months after discharge showed that the wounds of all patients healed well. One patient who underwent femoral artery ligation had calf amputation due to foot ischemic necrosis, and the rest of the patients regained normal walking ability. The pulsatile mass disappeared in inguinal region of all patients. B-ultrasound or CTA re-examination in 6 patients showed that the blood flow of femoral artery had good patency, and there was no pseudoaneurysm recurrence. Conclusions: Early debridement, tumor resection, and individualized artery treatment should be performed in patients with femoral artery pseudoaneurysm combined with infected wounds. Besides, proper drainage and personalized repair strategy should be conducted according to the wound condition to achieve a good outcome.

[Clinical efficacy and influencing factors of different modes of continuous negative pressure wound therapy on venous ulcer wounds of lower limbs].

Objective: To explore the clinical efficacy of different modes of continuous negative pressure wound therapy (NPWT) on venous ulcer wounds of lower limbs, and to analyze the influencing factors. Methods: From January 2018 to December 2019, 53 patients with venous ulcer of lower limbs who met the inclusion criteria and hospitalized in the Affiliated Hospital of Jiangnan University were recruited in this prospective randomized controlled study. According to the random number table, the patients were divided into single negative pressure therapy (SNPT) group (19 patients, 11 males and 8 females), cyclic alternating negative pressure therapy (CANPT) group (17 patients, 12 males and 5 females), and routine dressing change (RDC) group (17 patients, 10 males and 7 females), aged (47±11), (49±10), and (47±10) years respectively. After admission, patients in SNPT group were given continuous NPWT with the single negative pressure setting at -13.3 kPa, patients in CANPT group were also given continuous NPWT but with the cyclic alternating negative pressure setting from -16.0 to -10.7 kPa, while patients in RDC group were given dressing change with vaseline gauze soaked with iodophor. The wound healing rate was calculated on treatment day 7 and 14. Transcutaneous oxygen pressure (TcPO(2)) around the wound was detected by TcPO(2) meter before treatment and on treatment day 7 and 14. The wound exudate/drainage fluid was collected on treatment day 1, 4, 7, 10, and 14, with the pH value measured using a pH meter, and the volume of exudate/drainage fluid recorded. Before treatment and on treatment day 7 and 14, venous blood was collected to detect the serum levels of interleukin 1ß (IL-1ß), IL-6, tumor necrosis factor α(TNF-α), transforming growth factor-ß(1) (TGF-ß(1)), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Before treatment and on treatment day 7 and 14, wound exudates were collected for bacterial culture, and Visual Analogue Scale and Hamilton Anxiety Scale were used to evaluate the degree of wound pain and anxiety of patients respectively. The length of hospital stay and the total treatment cost were counted. Analysis of variance for repeated measurement, one-way analysis of variance, least significant difference test, Kruskal Wallis H test, Mann Whitney U test, chi-square test, Fisher's exact probability method test, and Bonferroni correction were used to analyze the data. According to the wound healing rate on treatment day 14, the efficiency of patients were divided into two grades of significant healing with wound healing rate≥70% and non significant healing with wound healing rate<70%. According to the two categories of wound healing rate as dependent variables, the levels of TcPO(2), IL-1ß, IL-6, TNF-α, TGF-ß(1), VEGF, bFGF levels and bacterial detection, wound pain and anxiety before treatment, wound exudate/drainage fluid volume and pH value on treatment day 1 were taken as covariates, and binary classification multifactor logistic regression analysis was used to analyze the risk factors of significant wound healing. Results: (1) On treatment day 7, the wound healing rate of patients in SNPT group was (33±10) %, which was significantly higher than (24±9) % of RDC group (P<0.05). On treatment day 14, the wound healing rates of patients in SNPT group and CANPT group were (71±15)% and (66±18)%, respectively, which were significantly higher than (45±19)% of RDC group (P<0.01). (2) Compared with those of RDC group, the TcPO(2) value around the wound of patients was significantly increased in SNPT group on treatment day 14 and in CANPT group on treatment day 7 and 14 (P<0.05 or P<0.01), the pH value of wound drainage fluid of patients was significantly decreased in SNPT group on treatment day 10 and 14 and in CANPT group on treatment day 7 and 14 (P<0.05), the volume of wound drainage fluid of patients was significantly reduced in SNPT group on treatment day 10 and 14 and in CANPT group on treatment day 7, 10, and 14 (Z=-4.060, -4.954, -2.413, -4.085, -4.756, P<0.05 or P<0.01), the serum levels of IL-1ß, IL-6, and TNF-α of patients were significantly decreased in SNPT group and CANPT group on treatment day 7 and 14 (P<0.01), the serum level of TGF-ß(1) of patients was significantly increased in CANPT group on treatment day 14 (P<0.05), the serum levels of VEGF and bFGF were significantly increased in SNPT group and CANPT group on treatment day 14 (P<0.01), the bacteria detection proportion of wound exudate, wound pain, and anxiety scores of patients were significantly decreased in SNPT group and CANPT group on treatment day 7 and 14 (P<0.01). Compared between the negative pressure therapy two groups, except the wound pain score of patients in CANPT group was significantly lower than that in SNPT group (P<0.01) on treatment day 7, the other indicators mentioned above were similar. (3) The length of hospital stay of patients in SNPT group was similar to that in CANPT group (P>0.05), which were significantly shorter than the time in RDC group (P<0.01). The total treatment cost of patients among the three groups was similar (F=1.766, P>0.05). (4) Before treatment, the serum levels of TNF-α and bFGF, TcPO(2) around the wound, and the degree of wound pain were risk factors for significant wound healing (odds ratio=1.109, 0.950, 1.140, 2.169, 95% confidence interval=1.012-1.217, 0.912-0.988, 1.008-1.290, 1.288-3.651, P<0.05 or P<0.01). Conclusions: Clinical application of continuous NPWT under single negative pressure mode and cyclic alternating negative pressure mode has a positive effect on improving the wound base and healing rate of venous ulcer of lower limbs. But cyclic alternating negative pressure mode is significantly more effective than single negative pressure mode in improving TcPO(2) around the wound, reducing wound pH value, reducing exudate volume and relieving pain. The serum levels of TNF-α and bFGF, TcPO(2) around the wound and the degree of wound pain were the risk factors that affect the wound healing significantly.

Bradykinin alleviates DR retinal endothelial injury by regulating HMGB-1/NF-κB pathway.

OBJECTIVE: Diabetic retinopathy (DR) is one of the most important complications of diabetes (DM) and the leading cause of blindness in adults. Bradykinin (BK) is involved in several pathophysiological processes, such as inflammation, pain, cell proliferation, and tumors. It plays a crucial role in corneal epithelial cells, corneal stromal cells, and fibroblasts. However, the role of BK in DR retinal endothelial injury remains unclear. PATIENTS AND METHODS: Human retinal microvascular endothelial cells (hRECs) were cultured in vitro and randomly divided into 3 groups, control group in which hRECs were cultured in normal glucose concentration, high glucose group in which hRECs were cultured in the presence of high glucose, and BK group in which hRECs were cultured in the presence of high glucose with 1 µM BK. The MTT assay (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) was used to detect cell proliferation. Caspase-3 activity was adopted to detect Caspase-3 activity in hRECs. The colorimetric method was selected to determine lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD) activity, and ROS content. Western blot was used to test HMGB-1/NF-κB and vascular endothelial growth factor (VEGF) expression changes. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the secretion of inflammatory factor tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß). RESULTS: In the presence of high glucose, hRECs cells proliferation was significantly reduced, Caspase-3 activity was enhanced, LDH and ROS levels were increased, SOD activity was declined with increased expression of HMGB-1, NF-κB, VEGF, as well as secretion of TNF-α and IL-1ß compared with control group (p < 0.05). BK significantly inhibited the proliferation of hRECs cells, enhanced Caspase-3 activity, decreased the content of LDH and ROS, increased SOD activity, reduced the expressions of HMGB-1 and NF-κB protein, attenuated the expression of VEGF, and restrained the secretion of TNF-α and IL-1ß compared with high glucose group (p < 0.05). CONCLUSIONS: BK can inhibit the growth and proliferation of retinal endothelial cells by regulating HMGB-1/NF-κB signaling pathway, attenuating oxidative stress and inflammation, thereby delaying DR development and progress.

Characterization and effects of miR-21 expression in esophageal cancer.

The aim of this study was to investigate the expression of miR-21 in esophageal cancer and the impact of miR-21 on apoptosis, invasion, and the expression of target genes in esophageal cancer cells. Fluorescence quantitative polymerase chain reaction analysis was used to detect the expression of miR-21 in human esophageal tissues, adjacent tissues, and an esophageal cancer cell line (TE-13). The antisense miR-21 oligonucleotide was generated commercially using the solid-phase chemical synthesis method. Transient transfection was used to transfect esophageal cancer cells (TE-13 antisense and TE-13 control cells). Flow cytometry and Transwell cell assays were used to detect the apoptosis and invasion of esophageal cancer cells, respectively. The western blot method was used to detect the expression of PTEN, PDCD4, and K-ras proteins. These analyses determined that mir-21 expression significantly increased in esophageal cancer tissues and in TE-13 cells, and that this phenomenon was not associated with staging or lymph node metastasis. The apoptosis rate of TE-13 control cells was lower than that of antisense TE-13 cells indicating an enhanced invasive ability. In tissues adjacent to esophageal cancer and in TE-13 antisense cells, the expression of PTEN and PDCD4 was found to be higher than that in the control group, whereas the expression of K-ras showed the opposite pattern. Together, these results suggest that miR- 21 might be involved in the development and metastasis of esophageal cancer, through interaction with its PDCD4 and K-ras target genes.

Association of miR-21 with esophageal cancer prognosis: a meta-analysis.

The present study aimed to explore the relationship between miRNA expression and survival in patients with esophageal cancer (EC) using meta-analysis. We searched PubMed, EMBASE, CNKI, Wanfang, and ISI Web of Science databases without time restrictions, and extracted relevant data, such as the name of first author, publication year, age, gender, number of case, etc. from the studies included. We calculated the pooled hazard ratios (HRs) using the RevMan 5.2 software. A total of five studies involving 504 subjects were included in the meta-analysis, with the purpose of analyzing the association of miRNA-21 expression with EC prognosis. The pooled HR of elevated versus decreased miR-21 expression in EC was 1.87 [95% confidence interval (CI): 1.37-2.55, P < 0.001], with elevated miR-21 expression being associated with poorer prognosis for patients with EC. Our results support a prognostic role for miR-21 in EC.

Differential characteristics of the early stage of lung inflammation induced by SARS-CoV Nucleocapsid protein related to age in the mouse.

OBJECTIVE: Severe acute respiratory syndrome (SARS) is an acute infectious disease of the respiratory system which has newly emerged. Interestingly, it appears to be a disease that predominantly affects adults while the mortality in children is extremely low. However, the pathogenesis of SARS in relation to different characteristics relevant to age remains unclear. MATERIAL AND METHODS: To better understand the role of cytokines in the immunopathological processes of SARS, weanling (4 weeks old), young (6 weeks old) and adult (10 weeks old) male BALB/C mice were inoculated intranasally with N-protein of SARS-CoV in this study. Serum or lung homogenate levels of some cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) along with acute injury lung index and histology were also analyzed. RESULTS: Histopathological analysis of adult male BALB/C mice after N-protein infection showed progressive inflammatory reactions, especially pulmonary edema, in accordance with a moderately (approximately 13%) elevated level of W/D ratio at 24 h. Although adult groups underwent a progressive lung inflammation in the acute phase accompanied by raised levels of TNF-alpha in serum, no significant changes in lung TNF-alpha level were reported simultaneously. Moreover, adult SARS infected BALB/c mice showed elevated levels of IFN-gamma while IFN-gamma levels in weanling and young groups had no obvious association with lung inflammation. CONCLUSION: Our study supports the observation that adult mice do have progressively greater immune reactions than weanling and adolescent ones over time. The relative immaturity of the immune system in weanlings may confer benefit leading to less impairment of lung function. However, the measurement of TNF-alpha and IFN-gamma levels was not indicative of the severity of lung injury at the early stage of disease.

A novel missense mutation L468Q of keratin 6a in pachyonychia congenita type 1.

BACKGROUND: Pachyonychia congenita is an autosomal dominant disorder that usually develops in early infancy. The major features of the syndrome are hypertrophic nail dystrophy, palmoplantar keratoderma and oral leucokeratosis, accompanied by other ectodermal defects, according to subtype. OBJECTIVE: To analyse the K6a gene mutation in a sporadic Chinese patient with pachyonychia congenita type 1 (PC-1) and to explore the relationship between the genotype and phenotype of PC-1. METHODS: Genomic DNA was extracted from peripheral blood of the patient with PC-1 and 100 unrelated controls. The whole coding region of K6a gene was amplified using long-range polymerase chain reaction (PCR); nested PCR was then used to amplify the mutation 'hot-spot' of the K6a gene. The PCR products were directly sequenced to detect the mutation. RESULTS: A novel missense mutation L468Q in the helix 2B domain of the K6a polypeptide was identified in the patient but not in the healthy individuals from the family and 100 unrelated control individuals. CONCLUSIONS: We describe this mutation for the first time, and provide further evidence that the helix boundary motif sequences of K6a are a mutation 'hot-spot'.

Is the effect of silicon on rice uptake of arsenate (AsV) related to internal silicon concentrations, iron plaque and phosphate nutrition?

Solution culture experiments were conducted to investigate the effects of silicon (Si) on arsenate (As(V)) uptake by rice. The addition of Si to the pretreatment or uptake solution significantly decreased shoot and root As concentrations (P<0.001 and P<0.05). The presence of Si in the pretreatment or uptake solution also significantly decreased shoot P concentrations (P<0.001). The data demonstrated that both internal and external Si inhibited the uptake of As and P. Results of As uptake kinetics showed that the mechanism of the effect of Si on arsenate uptake is not caused by direct competition for active sites of transporters with As. The effect of Si on As uptake was not entirely mediated through the effect of Si on P uptake. Although the addition of Si to pretreatment solutions still significantly decreased shoot and root As concentrations, the extent of reduction became smaller when rice roots were coated with iron plaque.

Role of salicylic acid in alleviating oxidative damage in rice roots (Oryza sativa) subjected to cadmium stress.

Time-dependent changes in enzymatic and non-enzymatic antioxidants, and lipid peroxidation were investigated in roots of rice (Oryza sativa) grown hydroponically with Cd, with or without pretreatment of salicylic acid (SA). Exposure to 50 microM Cd significantly decreased root growth, and activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), but increased the concentrations of H(2)O(2), malondialdehyde (MDA), ascorbic acid (AsA), glutathione (GSH) and non-protein thiols (NPT). However, pretreatment with 10 microM SA enhanced the activities of antioxidant enzymes and the concentrations of non-enzymatic antioxidants, but lowered the concentrations of H(2)O(2) and MDA in the Cd-stressed rice compared with the Cd treatment alone. Pretreatment with SA alleviated the Cd-induced inhibition of root growth. The results showed that pretreatment with SA enhanced the antioxidant defense activities in Cd-stressed rice, thus alleviating Cd-induced oxidative damage and enhancing Cd tolerance. The possible mechanism of SA-induced H(2)O(2) signaling in mediating Cd tolerance was discussed.

Arsenic sequestration in iron plaque, its accumulation and speciation in mature rice plants (Oryza sativa L.).

A compartmented soil-glass bead culture system was used to investigate characteristics of iron plaque and arsenic accumulation and speciation in mature rice plants with different capacities of forming iron plaque on their roots. X-ray absorption near-edge structure spectra and extended X-ray absorption fine structure were utilized to identify the mineralogical characteristics of iron plaque and arsenic sequestration in plaque on the rice roots. Iron plaque was dominated by (oxyhydr)oxides, which were composed of ferrihydrite (81-100%), with a minor amount of goethite (19%) fitted in one of the samples. Sequential extraction and XANES data showed that arsenic in iron plaque was sequestered mainly with amorphous and crystalline iron (oxyhydr)oxides, and that arsenate was the predominant species. There was significant variation in iron plaque formation between genotypes, and the distribution of arsenic in different components of mature rice plants followed the following order: iron plaque > root > straw > husk > grain for all genotypes. Arsenic accumulation in grain differed significantly among genotypes. Inorganic arsenic and dimethylarsinic acid (DMA) were the main arsenic species in rice grain for six genotypes, and there were large genotypic differences in levels of DMA and inorganic arsenic in grain.

Genetic heterogeneity in acrokeratosis verruciformis of Hopf.

BACKGROUND: Acrokeratosis verruciformis of Hopf (AKV) is a rare genodermatosis characterized by multiple flat-topped, flesh-coloured papules on the dorsa of hands and feet, and punctuate keratoses on the palms and soles. A mutation in the ATP2A2 gene has been shown to be associated with AKV and with Darier's disease (DD). OBJECTIVES: To explore the molecular aetiology of AKV and DD. METHODS: We investigated the clinical and histological information in two families and a sporadic case with AKV and one family and a sporadic case with DD in China. Mutation analysis of ATP2A2 was performed by PCR and direct sequencing, and genotyping and linkage analysis performed using six polymorphic microsatellite markers spanning the locus at 12q23-12q24 containing ATP2A2. RESULTS: Mutational analysis showed no mutation in ATP2A2 among the AKV patients, but we found two novel mutations (p.C318F and p.M719fs) in the DD patients. The genotyping and linkage analysis results revealed no linkage evidence of the locus at 12q23-12q24 in a large AKV family. CONCLUSIONS: Our findings provide evidence for the genetic heterogeneity of AKV and demonstrate that mutations in genes other than ATP2A2 are responsible for AKV in a proportion of the Chinese population.

Sequestration of As by iron plaque on the roots of three rice (Oryza sativa L.) cultivars in a low-P soil with or without P fertilizer.

A pot experiment was carried out in a greenhouse to investigate the sequestration of As in iron plaques on root surface of three rice (Oryza sativa L.) cultivars. Phosphate (P) fertilization increased both plant biomass and tissue P concentrations significantly, indicating that the soils used in this study was highly P-deficient. Results from this study confirmed that low P supply improved the formation of iron plaque on rice roots. As a consequence, arsenic (As) concentrations in DCB-extracts with no P addition were significantly higher than those with P fertilization. Arsenic was highly sequestrated in iron plaque; arsenic concentration in iron was up to nearly 120 mg kg(-1), while arsenic concentrations in roots were just several mg kg(-1). Both arsenic and phosphate concentrations in iron plaque were highly positively correlated with the amounts of iron plaque (DCB-extractable Fe). Contrary to normal understanding that increasing P supply could reduced As accumulation in plants, results from the present study showed that P fertilization did not inhibit the As uptake by plants (As accumulation in aboveground), which was probably due to the fact that iron plaque formation was improved under low P conditions, thus leading to more As sequestration in the iron plaque. Thus results obtained in this study indicated that the iron plaque may inhibit the transfer of As from roots to shoots, and thus alter the P-As interaction in plant As uptake processes.

Do iron plaque and genotypes affect arsenate uptake and translocation by rice seedlings (Oryza sativa L.) grown in solution culture?

The effects of Fe concentrations in the pretreatment solution on the induction of plaque and the differences between genotypes on arsenate uptake by and translocation within rice seedlings grown in nutrient solution in the greenhouse were investigated. After iron plaque on rice roots was induced in solutions containing 20, 40, 60, 80, and 100 mg Fe2+ l(-1), seedlings were transplanted into nutrient solution with 0.5 mg As l(-1). The formation of iron plaque was clearly visible as a reddish coating on the root surface after 12 h induction. Fe2+ concentrations in the pretreatment solution and 0.5 mg As l(-1) in the treatment solutions did not significantly affect rice growth. There was a significant correlation between the concentrations of Fe and As in iron plaque on the root surface for the three genotypes. About 75-89% of total As was concentrated in iron plaque (DCB-extracts). There were no significant differences in As concentrations in the roots between the three genotypes; however, As concentrations in shoots differed significantly between them. Arsenic concentrations in shoots were positively correlated with iron concentrations in the shoots. The results suggest that iron plaque may act as a 'buffer' for As in the rhizosphere.

Protective effect of ginsenoside Rg1 on MPP+-induced apoptosis in SHSY5Y cells.

The neuroprotective mechanism of Rg1 was studied in this paper by means of its obvious anti-apoptotic effect on human SHSY5Y cells. SHSY5Y cells were treated with MPP+ (1-methyl-4-phenyl-pyridinium) for 72 hours to induce apoptosis. During the apoptosis, production of reactive oxygen species (ROS), activation of c-Jun N-terminal kinase (JNK) and activation of caspase-3 were observed. The results showed that the signal transduction pathway of MPP+-induced apoptosis might be ROS to JNK, then to caspase-3. MPP+-induced apoptosis in SHSY5Y cells was obviously inhibited in both NAC (N-acetylcysteine) pretreated groups and Rg1 pretreated groups. Meanwhile, compared to that of the controls, our results showed decreased level of ROS, less JNK activity and lower expression of cleaved caspase-3 in pretreated NAC groups and in Rg1 pretreated groups. The protection by Rg1 might be mediated by removing of ROS. The removal of ROS might inhibit the activity of JNK and the expression of cleaved caspase-3. These results suggest that ginsenoside Rg1 may take effect through its anti-apoptotic activity in neurodegenerative diseases.

Cationic lipids enhanced cellular uptake and activity of bcl-2 antisense oligodeoxynucleotide G3139 in HL-60 cells.

AIM: To explore the effect of cationic lipid 1,3-di-oleoyloxy-2-(6-carboxy-spermyl)-propylamid (DOSPER) on cellular uptake and activity of bcl-2 antisense oligodeoxynucleotide G3139 in HL-60 cells. METHODS: The cell-associated mean fluorescence intensity and the percentage of cells with positive staining for Bcl-2 were measured by flow cytometry. The subcellular distribution of fluorescein isothiocyanate (FITC)-labeled G3139 was observed by fluorescence microscope and the bcl-2 mRNA level was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: (1) DOSPER increased cellular uptake of G3139 into HL-60 cells greatly. When DOSPER/G3139 (microg : microg) was 2 : 1, the uptake of G3139 reached top after treatment for 2 h and increased about 20 times compared with application of G3139 alone. In the presence of DOSPER, G3139 was localized in nucleus and cytosol with a bright spotted fluorescence staining. However, G3139 was localized in cytoplasm with faint fluorescence in the absence of DOSPER. (2) Cell-associated G3139 could be effluxed out of cells. After treated with G3139 in the presence of DOSPER for 4 h, the cell-associated G3139 could be fitted by C(t)=68.2e(-0.60) t+31.8e(-0.02) t (% of initial value), with a half-life of approximately 1.1 h. In the absence of DOSPER, the cell-associated G3139 could be fitted by C(t)=64.8e(-2.27) t+35.2e(-0.04) t, with a half-life of about 18 min. (3) In the presence of DOSPER, G3139 1.0 micromol/L specially reduced bcl-2 mRNA level, and Bcl-2 protein decreased from 97 % +/- 4 % to 70.6 % +/- 2.1 %. CONCLUSION: DOSPER enhanced the activity of G3139 and it might be attributed to increase of the cellular uptake and change of the subcellular distribution of G3139.

Protective effect of ginsenoside Rg1 on dopamine-induced apoptosis in PC12 cells.

AIM: To explore the possible molecular mechanism of exogenous dopamine-induced apoptosis in PC12 cells and the protective effect of ginsenoside Rg1. METHODS: Flow cytometric assay was used to quantify the apoptotic cells and measure the percentage of cells with positive Bcl-2 and Bax proteins. The morphology of apoptotic cells was evaluated by transmission electron microscope. DNA fragmentation was observed by gel electrophoresis. Caspase-3 activity was determined by fluorescent spectrofluorometer and the expressive bcl-2 and bax mRNA by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Dopamine 0.15, 0.30, 0.45, and 0.60 mmol/L induced PC12 cell apoptosis from 1.1 % +/- 0.4 % (control) to 41 % +/- 3 %, 46.4 % +/ -2.7 %, 53 % +/ -3 %, and 64.5 % +/- 2.7 %, respectively. After treatment with dopamine 0.45 mmol/L following pretreatment with Rg1 10 micromol/L for 24 h, the percentage of apoptotic cells and caspase-3 activity decreased from 53 % +/- 3 % and 683 +/- 8 (mean fluorescence intensity, MFI) to 1.9 % +/- 0.6 % and 325 +/- 5, and the percentage of cells with positive Bcl-2 protein increased from 14.3 % +/- 1.1 % to 25.9 % +/- 1.6 %, however, the percentage of cells with positive Bax protein decreased from 48 % +/- 3 % to 35 % +/- 3 %, compared with group treated with DA 0.45 mmol/L alone. CONCLUSION: Ginsenoside Rg1 protected PC12 cells against apoptosis by inhibiting the activation of caspase-3 and regulating the ratio of Bcl-2 to Bax protein.