BACKGROUND: There are so many experimental and clinical researches on levels of nitrogen monoxide (NO) in serum and endothelins in plasma of patients with stroke; however, ratio and significance between them are still unclear.OBJECTIVE: To observe dynamic changes of levels of NO in serum and endothelins in plasma of patients with stroke.DESIGN: Case-controlled observation.SETTING: Neurological Department and Clinical Neurological Laboratory of the Second Hospital affiliated to Suzhou University.PARTICIPANTS: A total of 216 patients with acute cerebral infarction including 133 males and 83 females and 112 cases with cerebral hemorrhage including 68 males and 44 females were selected from Neurological Department of the Second Hospital affiliated to Suzhou University from September 1999 to December 2001. Another 106 subjects including 63males and 43 females were regarded as healthy control group.METHODS: Contents of NO in serum and endothelins in plasma were measured on 328 patients with stroke and 106 healthy subjects in the courses of 1-3 days, 1, 2, 4, 8 and 12 weeks with nitrate reductase and radio-immunity methods, respectively.MAIN OUTCOME MEASURES: Contents of NO in serum and endothelins in plasma; ratio between NO in serum and endothelins in plasma (NO/endothelins).RESULTS: ① As compared with those in the control group, content of NO in serum of patients with cerebral infarction and cerebral hemorrhage was decreased and reached the lowest value during acute period (within 1-3 days), and then increased gradually and closed to the normal level at about 4 weeks. In addition, content of endothelins in plasma was increased obviously during acute period, reached the peak at 2 weeks, and then decreased gradually. The level was still high at stage of recovery and closed to normal value within 4-8 weeks. ② As compared with that in the control group, NO/endothelins was decreased in cerebral infarction group at the courses of 1-3 days (P ＜ 0.05), reached the lowest value at 1 week (P ＜ 0.001), and increased to the normal level at 2 weeks. Moreover,NO/endothelins was remarkably decreased in cerebral hemorrhage group at the courses of 1-3 days (P ＜ 0.001), reached the lowest value at 1week (P ＜ 0.001), and increased gradually. The changes of course were great and the level reached above normal value at 8 weeks. There was significant difference of dynamic changes of NO/endothelins between cerebral hemorrhage group and cerebral infarction group (P ＜ 0.05).CONCLUSION: NO and endothelins play an important role in onset and development of ischemic cerebrovascular disease and hemorrhagic cerebrovascular disease, and their contents are related to prognosis.

Objective To establish the standards for classification of urinary fistula after kidney transplantation. Methods From December 1993 to February 2009, 1313 cases of renal transplanta-tions were operated, out of which 102 cases of urinary fistulas occurred (7.8%). Based on the princi-ple of the urethral injury classification method, we divide urinary fistula into simple and complex clas-ses by the cause, location, and the severity of the disease. Results There were 81 cases (79.4%) of simple urinary fistulas, of those 76 cases were ureteral end necrosis,4 cases were due to ureter blad-der anastomosis suture,1 case was anastomotic problem caused by wound infection. There were 21 ca-ses(20.6%) of complex urinary fistulas, of these 2 cases had fistulas at renal pelvis, 11 cases at ure-ter-bladder interface and 6 cases had ureteral necrosis longer than 2 cm. For the 81 cases urinary fistu-las patients, 34 patients conservative treatments were cured and 47 patients need surgeries. For all complex urinary fistulas need surgeries: 11 cases had surgery once, 5 cases had 2 times, 3 cases had 3 times and 2 cases had 4 times. Among the 2 groups, three patients (2.9%) died of urinary fistulas which led to severe lung infection. Conclusions A "Five Steps Procedure" could be used for diagno-sis and treatment of post renal transplantation fistula. The urinary fistulas are divided into simple and complex types after renal transplantation. This provides a guidance for the best choice of treatment.

1 313 patients who received renal transplantations at Department of Urology, Research Institute of Field Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from December 1993 to October 2008 were selected in the experiment. Urinary fistula occurred in 68 patients of them after renal transplantation. In order to make diagnosis more standard, 68 patients was classified in accordance with diagnostic classification standards after renal transplantation. The 68 patients were divided into simple and complex urinary fistulas in accordance with lesion degree. They were divided into low, high and multiple fistulas in accordance with the position and etiology. 47 (69.1%) of 68 cases were simple urinary fistulas: 42 cases were because of terminal ureteral necrosis; 4 cases were because the anastomosis was mended unsuitably; 1 case was because of poor healing of anastomosis due to infections. 21(30.9 %) cases were complex urinary fistulas. The position of orificium fistula: orificium fistula located at renal pelvis, ureter and anastomosis were 2, 2 and 11 cases, respectively. 6 cases had ureteral necrosis longer than 2 cm. The times of repair: 11 cases had 1 time, 5 cases had 3 times, 3 cases had 3 times and 2 cases had 4 times. 2 cases (2.9%) died because of severe pulmonary infection caused by urinary fistula. Result suggests that there are two advantages of dividing urinary fistula into the simple and complex types after renal transplantation: one is that the diagnosis of urinary fistula is more carefully and standardized, and the other is that doctors can make the best choice for treatment in order to get the best efficacy.

Binding Sites ; Cysteine Endopeptidases ; metabolism ; Humans ; Isoxazoles ; chemistry ; pharmacology ; Protease Inhibitors ; chemistry ; metabolism ; pharmacology ; Protein Structure, Tertiary ; Pyrrolidinones ; chemistry ; pharmacology ; Rhinovirus ; drug effects ; SARS Virus ; drug effects ; enzymology ; Severe Acute Respiratory Syndrome ; virology ; Viral Proteins ; antagonists & inhibitors ; metabolism

BACKGROUNDTo construct recombinant adenoviral vector carrying Smad3D or Smad7 by a simplified means.

METHODSBased on AdEasy System, adenoviral backbone plasmid vector and shuttle vector carrying the gene of interest were transferred into E.coli BJ5183 by chemical transformation methods in special order. The homologous recombination was performed.

RESULTSRecombinant adenoviral vector pAd-Smad3D and pAd-Smad7 were constructed successfully, which were confirmed by restriction enzyme digesting.

CONCLUSIONSRecombinant adenoviral vector may be constructed quickly and efficiently in E.coli by sequential chemical transformation methods.

Objective To analyze the clinical values of super early enteral nutrition combined with microecopharmaceutics and delayed enteral nutrition on patients with severe acute pancreatitis. Methods Clinical data of thirty patients diagnosed as severe acute pancreatitis in our emergency department during January 2013 and December 2017 were reviewed retrospectively. Patients were divided into the treatment group (n=15, patients given enteral nutrition combined with microecopharmaceutics within 24 h after admission) and the control group (n=15, patients given delayed enteral nutrition after 48 h of admission). Two weeks after the treatment, the serum variables of C-reactive protein, total protein, albumin, recovery time of urine and blood amylase, length of hospital stay and APACHE Ⅱ score were compared between the two groups by using paired samples t test. Results The C-reactive protein [(46.7±13.1) mg/L vs. (190.72±19.3) mg/L, t=10.4, P<0.01] and APACHE Ⅱ score [(7.2±1.9) vs.(9.3±2.4),t=2.7,P<0.05] of the treatment group were significantly lower than those in the control group. The total protein [(58.1±6.3)g/L vs.(52.6±5.4)g/L, t=2.5, P<0.05] and albumin [(29.9±3.2)g/L vs.(22.0±2.8)g/L, t=7.12, P<0.01] of the treatment group were significantly higher than those in the control group. The recovery time of urine amylase [(13.2±2.1)d vs.(18.7±3.9)d, t=4.9, P<0.01] and blood amylase [(7.5±3.0)d vs.(11.1±3.4)d, t=3.1, P<0.01], and length of hospital stay[(14.9±4.5)d vs.(27.1±5.3)d, t=6.9, P<0.01] were significantly shorter in the treatment group compared with those in the control group. Conclusions Ultra-early enteral nutrition combined with microecopharmaceutics can shorten the length of hospital stay of patients with severe acute pancreatitis, and is safe and effective.

Objective:To discusses effectiveness of CTA in the classification of first plantar metatarsal artery and its application value in thumb reconstruction.Methods:Thirty-six cases who underwent thumb reconstruction with free second toe or hallucis flap between December, 2015 and December, 2018 were retrospectively analyzed. Among these cases, 22 cases were injured by machine stranding, 7 cases by heavy objects, and 7 cases were injured by rolling. Exact first plantar metatarsal arteries of all these cases were evaluated by using CTA preoperatively. And compared with intraoperative findings at the donor sites. Free first or second toe flap for thumb reconstruction were designed preoperatively based on branching pattern of first plantar metatarsal arteries.Results:The origin, course, 3-dimensional (3D) anatomical relationship with surrounding tissues and branching pattern of all these feet first plantar metatarsal arteries of 35 cases (70 feet)were well displayed in CTA images, and 1 case (2 feet) were showed poor vascular continuity and artifacts in CTA (2.78%). According to the branching pattern of first plantar metatarsal arteries, 29 cases (58 feet, 80.56%) were ramifying type, 5 cases (10 feet, 13.88%) were main trunk type, and 1 case (2 feet, 2.78%) were tiny branch type. Preoperative CTA images and intraoperative findings at the donor site of 35 cases were remarkably consistent. According to CTA images, 27 cases underwent thumb reconstruction with hallucis flap, 8 cases underwent thumb reconstruction with second toe, and 1 case of poor vascular continuity and artifacts in CTA underwent thumb reconstruction with hallucis flap eventually. All these cases were followed-up for 6-24 (average 12) months, and all reconstructed thumbs survived. The clinical outcomes of all these reconstructed thumbs were good with satisfactory appearance, sensory recovery, excellent motion. The donor feet of all cases recovered well.Conclusion:High-quality 3D images of first plantar metatarsal arteries could be obtained by CTA, allowing preoperative assessment of blood supply and planning of donor site. Therefore, success rate of reconstructed operation could be improved with low disability rate of donor site.

Objective To investigate the target blood pressure level of restrictive fluid resuscitation in patients with traumatic hemorrhagic shock. Methods Sixty patients with traumatic hemorrhagic shock admitted to the First Affiliated Hospital of Bengbu Medical College from January 2016 to December 2018 were enrolled. All patients were resuscitated with sodium acetate ringer solution after admission. According to the difference of mean arterial pressure (MAP) target, the patients were divided into low MAP (60 mmHg ≤ MAP < 65 mmHg, 1 mmHg = 0.133 kPa), middle MAP (65 mmHg ≤ MAP < 70 mmHg) and high MAP (70 mmHg ≤ MAP < 75 mmHg) groups by random number table using the admission order with 20 patients in each group. Those who failed to reach the target MAP after 30-minute resuscitation were excluded and supplementary cases were deferred. The restrictive fluid resuscitation phase was divided into three phases: before fluid resuscitation, liquid resuscitation for 30 minutes and 60 minutes. The most suitable resuscitation blood pressure level was further speculated by monitoring the inflammatory markers and hemodynamics in different periods in each group of patients. Pearson correlation analysis was used to detect the correlation of variables. Results Before fluid resuscitation, there was no significant difference in hemodynamics or expressions of serum cytokines among the three groups. Three groups of patients were resuscitated for 30 minutes to achieve the target blood pressure level and maintain 30 minutes. With the prolongation of fluid resuscitation time, the central venous pressure (CVP), cardiac output (CO) and cardiac index (CI) were increased slowly in the three groups, and reached a steady state at about 30 minutes after resuscitation, especially in the high MAP group and the middle MAP group. The expressions of serum inflammatory factors in the three groups were gradually increased with the prolongation of fluid resuscitation time. Compared with the low MAP group and the high MAP group, after 30 minutes of resuscitation the middle MAP group was superior to the other two groups in inhibiting the expressions of pro-inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and promoting anti-inflammatory factors IL-10 [TNF-α mRNA (2-ΔΔCt):0.21±0.13 vs. 0.69±0.34, 0.57±0.35; IL-6 mRNA (2-ΔΔCt): 0.35±0.31 vs. 0.72±0.39, 0.59±0.42; IL-10 mRNA (2-ΔΔCt): 1.25±0.81 vs. 0.61±0.46, 0.82±0.53; all P < 0.05], but there was no significant difference in promoting the expression of IL-4 mRNA among three groups. At 60 minutes of resuscitation, compared with the low MAP group and the high MAP group, the middle MAP group could significantly inhibit the expressions of TNF-α, IL-6 and promote IL-10 [TNF-α mRNA (2-ΔΔCt): 0.72±0.35 vs. 1.05±0.54, 1.03±0.49; IL-6 mRNA (2-ΔΔCt): 0.57±0.50 vs. 1.27±0.72, 1.01±0.64; IL-10 mRNA (2-ΔΔCt): 1.41±0.90 vs. 0.81±0.48, 0.94±0.61; all P < 0.05]. Compared with the high MAP group, the middle MAP group had significant differences in promoting the expression of IL-4 mRNA (2-ΔΔCt: 1.32±0.62 vs. 0.91±0.60, P < 0.05). There was no significant difference in serum cytokine expressions at different time points of resuscitation between the low MAP group and the high MAP group (all P > 0.05). Correlation analysis showed that there was a strong linear correlation between MAP and mRNA expressions of TNF-α, IL-6, IL-10 in the middle MAP group (r value was 0.766, 0.719, 0.692, respectively, all P < 0.01), but had no correlation with IL-4 (r = 0.361, P = 0.059). Fitting linear regression analysis showed an increase in 1 mmHg per MAP, the expression of TNF-α mRNA increased by 0.027 [95% confidence interval (95%CI) = 0.023-0.031, P < 0.001], IL-6 mRNA increased by 0.021 (95%CI = 0.017-0.024, P < 0.001), and IL-10 mRNA increased by 0.049 (95%CI = 0.041-0.058, P < 0.001). Conclusions When patients with traumatic hemorrhagic shock received restrict fluid resuscitation at MAP of 65-70 mmHg, the effect of reducing systemic inflammatory response and improving hemodynamics is better than the target MAP at 60-65 mmHg or 70-75 mmHg. It is suggested that 65-70 mmHg may be an ideal target MAP level for restrictive fluid resuscitation.

Objective To evaluate the value of 3D printing technology to assist in the repair of severe damage to the radial side hand.Methods From March,2014 to September,2016,there were 9 cases suffered from varying degrees of metacarpal and phalangeal bones injuries with soft tissue destruction at the radial side of the hand.The donor and recipient areas were scanned by 64 row spiral CT before operation.Three dimensional images were obtained by CTA 3D reconstruction technology.Preoperative plan was designed by using Mirror image technology and Virtual surgery aided design technique of Mimics software virtual surgery aided design system.Nine cases were treated with first toe toenail flap with dorsal foot flap combined with second phalangeal bone transplantation to reconstruct thumb and anterolateral thigh perforator flap to repair the wound.The cutting area of the dorsalis pedis flap and anterolateral thigh perforator flaps ranged from 4.0 cm × 5.5 cm to 6.5 cm × 9.5 cm and from 9.5 cm ×17.0 cm to 12.0 cm × 25.5 cm,respectively.Postoperative followed-up was performed to evaluate the effect of the operation.Results All 9 thumbs and flaps survived after the operation.Postoperative followed-up time ranged from 8 to 16 months.The appearance of reconstructed thumb was close to normal,and the two-point discrimination was from 8.0 to 11.0 mm.According to the evaluation criteria of upper limb function of the Chinese Academy of Medical Sciences,7 cases were excellent and 2 cases were good.The survival flaps had soft texture,good blood circulation and satisfactory appearance at the repaired site.Conclusion 3D printing technology assists thumb reconstruction and wound repair for severe damage to the radial side of the hand with good clinical results.

OBJECTIVE: To investigate the effect of calenduloside E on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells and explore the underlying molecular mechanism. METHODS: CCK-8 assay was used to examine the effect of different concentrations of calenduloside E (0-30 μg/mL) on the viability of RAW264.7 cells. The release of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells in response to pretreatment with 6, 8, and 10 μg/mL calenduloside E for 2 h followed by stimulation with 100 ng/mL LPS was detected using enzyme-linked immunosorbent assay (ELISA). The expression levels of iNOS and COX-2 and the activation of JAK-stats, MAPKs and NF-кB signaling pathways in the treated cells were determined using Western blotting. A reactive oxygen species (ROS) detection kit was used to detect ROS production in the cells, and the nuclear translocation of the transcription factor stat3 was observed by laser confocal microscopy. RESULTS: Calenduloside E below 20 μg/mL did not significantly affect the viability of RAW264.7 cells. Calenduloside E dose-dependently decreased the expression levels of iNOS and COX-2 induced by LPS, inhibited LPS-induced release of TNF-α and IL-1β, and suppressed LPS-induced JAK1-stat3 signaling pathway activation and stat3 nuclear translocation. Calenduloside E also significantly reduced ROS production induced by LPS in RAW264.7 cells. CONCLUSIONS Calenduloside E inhibits LPS-induced inflammatory response by blocking ROS-mediated activation of JAK1-stat3 signaling pathway in RAW264.7 cells.
Animals ; Lipopolysaccharides ; Mice ; NF-kappa B ; Oleanolic Acid ; analogs & derivatives ; RAW 264.7 Cells ; Reactive Oxygen Species ; Saponins ; Signal Transduction