[Efficacy analysis of retroperitoneal laparoscopic selective renal artery branch occlusion and nephron sparing surgery for ≥ T1b stage renal carcinoma].

Objective: To investigate the safety and efficacy of retroperitoneal laparoscopic selective renal artery branch occlusion with nephron sparing surgery in patients with renal carcinoma of stage ≥ T1b. Methods: From July 2016 to September 2020, 35 patients with renal cancer ≥T1b underwent retroperitoneoscopic nephron sparing surgery in the First Affiliated Hospital of Shenzhen University. The surgical methods were retroperitoneoscopic nephron sparing surgery with total renal artery occlusion (group A) or selective renal artery branch occlusion (group B). Operation time, heat ischemia time, blood transfusion rate, positive margin rate, intraoperative blood loss, postoperative complications and length of hospital stay were compared between the two groups, and the total glomerular filtration rate (GFR) and the single-nephron glomerular filtration rate (sGFR) of the offected kidneys were compared between the two groups before, 3 months after and 12 months after surgery. Results: Among the 35 patients, 19 were male and 16 were female, aged (55.7±8.4) years and the body mass index is (24.6±3.1) kg/m2. The tumor diameter was (54.7±10.3) mm. The difference was statistically significant of operative time between group A and B [(103.5±14.3) vs (123.2±14.1) min,P=0.003]. There were no significant differences in thermal ischemia time, blood transfusion rate, positive margin, intraoperative blood loss, incidence of postoperative complications and length of hospital stay between the two groups (all P>0.05). The decrease of renal sGFR in the group A was significantly higher than group B at 3 months and 12 months after surgery [(23.1±3.6) vs (29.1±7.1) ml/min;(25.9±4.7) vs (30.7±7.2),both P<0.05]. Conclusion: Retroperitoneal laparoscopic selective renal artery branch occlusion and neon-sparing surgery for patients with ≥ T1b stage renal carcinoma is a safe and effective surgical method, which can well protect the renal function of patients in the early postoperative stage without increasing intraoperative blood loss and postoperative complications.

LncRNA MEG3 participates in neuronal cell injury induced by subarachnoid hemorrhage via inhibiting the Pi3k/Akt pathway.

OBJECTIVE: To explore the effect of LncRNA MEG3 in the subarachnoid hemorrhage (SAH) and its underlying mechanism. PATIENTS AND METHODS: The expressions of lncRNA MEG3 in SAH patients and animal model were detected by quantitative real-time PCR (qRT-PCR). After LncRNA MEG3 was overexpressed in neurons by lentivirus, viability and apoptosis abilities were detected by cell counting kit-8 (CCK-8) assay, flow cytometry, and TUNEL assay, respectively. The apoptosis-related genes and Pi3k/Akt pathway-related proteins were further detected by a Western blot. RESULTS: The expressions of lncRNA MEG3 in SAH patients were remarkably higher than normal controls, which were positively correlated with SAH severity. After lncRNA MEG3 overexpression, neuronal cell activity was decreased and cell apoptosis was increased. Moreover, the expressions of Bax, p53, and cleaved Caspase-3 were increased, whereas the expression of Bcl-2 and Pi3k/Akt pathway-related proteins were decreased after lncRNA MEG3 overexpression. CONCLUSIONS: LncRNA MEG3 is up-regulated in SAH, which may promote SAH-induced neuronal cell injury via inhibition of the Pi3k/Akt pathway.

[Influence of different inner dressings in negative-pressure wound therapy on escharectomy wound of full-thickness burn rabbits].

Objective: To explore the influence of different inner dressings in negative-pressure wound therapy (NPWT) on escharectomy wound of full-thickness burn rabbits. Methods: Eighteen Japanese white rabbits were inflicted with full-thickness burn on unilateral back. They were divided into polymer dressing group (PD), biological dressing group (BD), and silver biological dressing group (SBD), according to the random number table, with 6 rabbits in each group. On 3 days post burn, the wounds were performed with escharectomy, and then wounds of rabbits in group PD were covered with polyurethane foam. Wounds of rabbits in group BD were covered with porcine acellular dermal matrix (ADM) and wounds of rabbits in group SBD were covered with silver porcine ADM. Then continuous NPWT was performed on rabbits of the three groups for 7 days. Immediately after surgery and on post surgery day (PSD) 7, general observation of wound was conducted and tissue around the wound was harvested for determination of dry to wet weight ratio. The content of bacteria was counted and the content of tumor necrosis factor α (TNF-α), interleukin-1ß (IL-1ß), and IL-6 in wound was determined by enzyme-linked immunosorbent assay. Fibroblasts in wound were counted after Masson staining and number of microvessels was counted after CD31 antibody immunohistochemical staining. Data were processed with analysis of variance for repeated measurement, LSD-t test, paired samples t test, and Bonferroni correction. Results: (1) Immediately after surgery, there was no granulation tissue in basal wound of rabbits in the three groups, with rich blood supply and obvious edema. On PSD 7, much granulation tissue was found in basal wound of rabbits in the three groups, with no or mild edema and no obvious redness and swelling in wound edge. (2) There were no significant differences in dry to wet weight ratios of tissue around the wound among and within the three groups immediately after surgery and on PSD 7 (with F values respectively 0.70 and 0.09, t values from 0.17 to 0.52, P values above 0.05). (3) Immediately after surgery, the content of bacteria in wounds of rabbits in groups PD, BD, and SBD was respectively (603.0±146.0) ×10(4,) (573.0±63.0) ×10(4,) and (590.0±100.0)×10(4) colony-forming unit (CFU)/g, with no significant difference among them (F=0.13, P>0.05). On PSD 7, the content of bacteria in wounds of rabbits in groups PD, BD, and SBD were respectively (5.4±0.8) ×10(4,) (4.6±0.9) ×10(4,) and (3.5±0.9)×10(4) CFU/g. Among them, the content of bacteria in wounds of rabbits in group SBD was lower than that in groups PD and BD, respectively (with t values respectively 3.78 and 2.29, P<0.05 or P<0.01). The content of bacteria in wounds of rabbits in the three groups on PSD 7 was decreased compared with that immediately after surgery (with t values from 10.05 to 21.81, P values below 0.01). (4) There was no significant difference in content of TNF-α, IL-1ß, and IL-6 in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values from 0.10 to 1.89, P values above 0.05). The content of TNF-α in wounds of rabbits in the three groups on PSD 7 was significantly higher than that immediately after surgery (with t values from 2.93 to 5.01, P<0.05 or P<0.01). (5) There was no significant difference in amount of fibroblasts in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values respectively 0.01 and 0.81, P values above 0.05). The amount of fibroblasts in wounds of rabbits in the three groups on PSD 7 was larger than that immediately after surgery (with t values from 4.78 to 11.58, P values below 0.01). (6) There was no significant difference in number of microvessels in wounds of rabbits in the three groups immediately after surgery and on PSD 7 (with F values respectively 2.42 and 2.49, P values above 0.05). The number of microvessels in wounds of rabbits in the three groups on PSD 7 was larger than that immediately after surgery (with t values from 7.17 to 11.14, P values below 0.01). Conclusions: SBD is better at inhibiting the growth of bacteria. PD, BD, and SBD have almost the same effects on reducing tissue edema and inflammatory reaction, and on promoting the accumulation of collagen fibers and tissue vascularization.

Quantifying Sarcopenia Reference Values Using Lumbar and Thoracic Muscle Areas in a Healthy Population.

BACKGROUND: Sarcopenia is defined as the loss of skeletal muscle mass and function associated with aging. Muscle mass can be reliably and accurately quantified using clinical CT scans but reference measurements are lacking, particularly in healthy US populations. METHODS: Two-phase CT scans from healthy kidney donors (age 18-40) at the University of Michigan between 1999-2010 were utilized. Muscle mass was quantified using two thoracic and two lumbar muscle cross-sectional area (CSA) measures. Indexed measurements were computed as area divided by height-squared. Paired analyses of non-contrast and contrast phases and different Hounsfield Unit (HU) ranges for muscle were conducted to determine their effect on CSA muscle measures. We report the means, standard deviations, and 2SD sarcopenia cutoffs from this population. RESULTS: Healthy population CSA (cm2) cutoffs for N=604 males/females respectively were: 34.7/20.9 (T12 Dorsal Muscle), 91.5/55.9 (T12 Skeletal Muscle), 141.7/91.2 (L3 Skeletal Muscle), 23.5/14.3 (L4 Total Psoas Area), and 23.4/14.3 (L4 Psoas Muscle Area). Height-indexed CSA (cm2/m2) cutoffs for males/females respectively were: 10.9/7.8 (T12 Dorsal Muscle), 28.7/20.6 (T12 Skeletal Muscle), 44.6/34.0 (L3 Skeletal Muscle), 7.5/5.2 (L4 Total Psoas Area), and 7.4/5.2 (L4 Psoas Muscle Area). We confirmed that a mask of -29 to 150 HU is optimal and shows no significant difference between contrast-enhanced and non-contrast CT scan CSA measurements. CONCLUSIONS: We quantified reference values for lumbar and thoracic muscle CSA measures in a healthy US population. We defined the effect of IV contrast and different HU ranges for muscle. Combined, these results facilitate the extraction of clinically valuable data from the large numbers of existing scans performed for medical indications.

[Effects of different fluid resuscitation program on renal function in swine during shock stage of severe burn].

Objective: To explore the effects of different fluid resuscitation program on renal function in swine during shock stage of severe burn. Methods: Twenty-four Guangxi Bama miniature swine were inflicted with 40% total body surface area on the back, and then they were divided into four groups according to the random number table, with 6 swine in each group. At post injury hour (PIH) 2, swine in succinylated gelatin group (S), hydroxyethyl starch group (H), and allogeneic plasma group (A) were respectively resuscitated with succinylated gelatin, hydroxyethyl starch 130/0.4, and plasma according to burn shock " domestic general" resuscitation formula, and swine in Parkland group (P) were resuscitated with lactated Ringer's solution according to Parkland formula. Hemodynamic indexes including heart rate, blood pressure, urine volume, pulmonary capillary wedge pressure, and central venous pressure before injury, at the first and second PIH 24 were recorded. The volume of resuscitation fluid was calculated at the first and second PIH 24. Blood and urine samples were collected before injury and at PIH 4, 8, 24, and 48, and then serum creatinine and urea nitrogen were detected by automatic biochemical analyzer, urine microalbumin and urine creatinine were detected by automated urine analyzer and the ratio of which was calculated. The renal tissue of swine in each group was obtained at PIH 48, and the pathologic changes were observed by optical microscopy and electron microscope. Data were processed with analysis of variance of repeated measurement, one-way analysis of variance, and LSD test. Results: (1) The hemodynamic indexes of swine in each group were similar before injury and at the first and second PIH 24 (with P values above 0.05). Compared with those before injury, except that the heart rate of swine in group A had no significant change at the first PIH 24 (P>0.05), the heart rate of swine in each group was significantly increased at the first and second PIH 24 (with P values below 0.01); except that the systolic blood pressure of swine in group P was significantly increased at the first and second PIH 24 (P<0.05 or P<0.01), there were no significant changes of systolic blood pressure and diastolic blood pressure of swine in each group at the first and second PIH 24 (with P values above 0.05); except that urine volume of swine in groups S and A was significantly decreased at the first PIH 24 (P<0.05 or P<0.01), there were no significant change of urine volume of swine in each group at the first and second PIH 24 (with P values above 0.05); pulmonary capillary wedge pressure and central venous pressure of swine in each group were significantly increased at the first and second PIH 24 (P<0.05 or P<0.01). (2) Compared with that in group A, the volume of resuscitation fluid of swine in groups S and H had no significant change in the first and second PIH 24 (with P values above 0.05), while it was significantly increased in group P in the first PIH 24 and significantly decreased in the second PIH 24 (with P values below 0.05). (3) Compared with those in group A, except that serum creatinine of swine in group H was significantly increased at PIH 24 and significantly increased in group P at PIH 4, 8, 24, and 48, urea nitrogen of swine in group P was significantly decreased at PIH 4 and 8 and significantly increased at PIH 48, the ratio of urine microalbumin to urine creatinine of swine in group P was significantly increased at PIH 8, 24, and 48 (P<0.05 or P<0.01), serum creatinine, urea nitrogen, and the ratio of urine microalbumin to urine creatinine of swine in each group had no significant change at each time point (with P values above 0.05). Serum creatinine of swine in group P was (125±16) µmol/L at PIH 24, which was significantly higher than that before injury [(75±13) µmol/L, P<0.05]. Urea nitrogen of swine in group S was (2.90±1.17) µmol/L at PIH 48, which was significantly lower than that before injury [(4.60±0.47) µmol/L, P<0.05]; urea nitrogen of swine in group H was (4.82±0.82) µmol/L at PIH 4, which was significantly higher than that before injury [(3.80±0.73) µmol/L, P<0.05]; urea nitrogen values of swine in group A were respectively (4.80±0.33), (4.92±0.35), and (2.60±0.27) µmol/L at PIH 4, 8, and 48, while those at PIH 4, 8 were significantly higher and at PIH 48 was significantly lower than the value before injury [(3.93±0.32) µmol/L, with P values below 0.01]. The ratios of urine microalbumin to urine creatinine of swine in group P were respectively (106.7±16.4) and (171.6±36.9) mg/mmol at PIH 24 and 48, which were significantly higher than the ratio before injury [(59.0±3.0) mg/mmol, with P values below 0.01]. The serum creatinine, urea nitrogen, and the ratio of urine microalbumin to urine creatinine of swine in each group at the other time points were similar to those before injury (with P values above 0.05). (4) The renal tissue of swine in the four groups had no obvious pathological change. Conclusions: According to the renal function results, fluid resuscitation with electrolyte and colloids are better than with lactated Ringer's solution in swine during shock stage of burn injury, while natural colloids and succinylated gelatin have similar effects, and both are superior to hydroxyethyl starch 130/0.4.

[Effects on wound bed of deep burn following eschar excision with different wound management in rabbits].

OBJECTIVE: To compare the effects on wound bed of deep burn following eschar excision with different wound management in rabbits. METHODS: Eighteen full-thickness burns models of Japanese white rabbits were established. They were randomly divided into 3 groups of traditional dressing, biological dressing and negative pressure wound therapy (NPWT) (n=6 each), according to the random number table. Eschar excision was performed three days later. The wound bed was observed and wound tissue was harvested for counting the quantity of bacteria, tissue dry wet ratio, measuring the level of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1ß and IL-6, the amount of collagen fibers and the microvessel density instantly and again seven days later. Statistical analyses were performed. RESULTS: The NPWT group was better than other groups by observing the wound bed. The quantity of bacteria of traditional dressing group, biological dressing group and NPWT group at the time point of seven days after escharectomy turned out to be (9.4±1.5)×10(4,) (8.1±2.7)×10(4,) (3.9±0.7)×10(4) cfu/g, the NPWT group was significantly lower than traditional dressing group and biological dressing group (both P<0.05), and all lower than that at the time point of the day when escharectomy was performed (576.9±169.5)×10(4,) (589.9±99.6)×10(4,) (583.0±160.4)×10(4) cfu/g ( all P<0.05). There were no statistically significant differences among three groups at two time points in tissue dry wet ratio (all P>0.05). The IL-6 of biological dressing group was higher than that of traditional dressing group at the time point of seven days after the eschar excision was performed[(94±10) vs (76±8) ng/L, P<0.05]. The amount of collagen fibers of three group at the time point of seven days after escharectomy turned out to be (60±9), (55±12), (77±17). The NPWT group was significantly higher than traditional dressing group and biological dressing group (P<0.05), and all higher than that at the time point of the day when escharectomy was performed[(39±6), (39±11), (38±6)](all P<0.05). The microvessel density of three groups at the time point of seven days after escharectomy turned out to be (42±6), (53±4), (82±10). The NPWT group was higher than that of the other two groups, and biological dressing group was higher than that of traditional dressing group (all P<0.05). The biological dressing group and NPWT group were both higher than that of the day when the eschar excision was performed (36±5) and (36±5) (P<0.05). CONCLUSIONS: NPWT is the optimal selection for wound to inhibit the growth of bacteria, promote the accumulation of collagen and tissue vascularization. But these managements have similar effects on reducing tissue edema and inflammatory reaction.

Lipopolysaccharide binding protein inhibitory peptide protects against acetaminophen-induced hepatotoxicity.

Acetaminophen (APAP)-induced liver injury remains the main cause of acute liver failure in the United States. Our previous work demonstrated that LPS binding protein (LBP) knockout mice are protected from APAP-induced hepatotoxicity. LBP is known to bind avidly to LPS, facilitating cellular activation. In this study, we sought to specifically inhibit the interaction between LBP and LPS to define the role of this interaction in APAP-induced liver injury. The peptide LBPK95A was able to inhibit LBP-mediated LPS activation of RAW 267.4 cells in a dose-dependent manner in vitro. In vivo, C57Bl/6 mice were treated with either LBPK95A or vehicle control concurrently with the administration of APAP (350 mg/kg). Mice treated with LBPK95A had significantly lower serum aspartate aminotransferase and alanine aminotransferase levels. Morphometric analysis of the liver tissue showed significantly less liver injury in mice treated with LBPK95A. To assess whether the LBPK95A altered glutathione depletion and APAP metabolism, we measured total glutathione levels in the liver after APAP. We found no difference in the glutathione levels and APAP-adduct formation between LBPK95A vs. vehicle control both at baseline and after APAP. In conclusion, our results support the hypothesis that LBP-induced liver injury after APAP is due to its ability to mediate activation by endogenous LPS. Our results suggest that blocking LBP-LPS interactions is a potential therapeutic avenue for the treatment of APAP-induced liver injury.

Investigation of valence orbitals of propene by electron momentum spectroscopy.

The binding energy spectra and momentum distributions of all valence orbitals of propene were studied by electron momentum spectroscopy (EMS) as well as Hartree-Fock and density functional theoretical calculations. The experiment was carried out at impact energies of 1200 eV and 600 eV on the state-of-the-art EMS spectrometer developed at Tsinghua University recently. The experimental momentum profiles of the valence orbitals were obtained and compared with the various theoretical calculations. Moreover, the experiment with a new analysis method presents a strong support for the correct ordering of the orbital 8a' and 1a'', i.e., 9a' < 8a' < 1a'' < 7a'.

Direct observation of distorted wave effects in ethylene using the (e,2e) reaction.

We report here the direct measurements of electron momentum distributions for ethylene using the (e,2e) reaction at different impact energies from 400 to 2400 eV. The "turn up" effects in the (e,2e) cross sections of the 1b(3g) orbital compared with the plane-wave impulse approximation calculations were observed at low and high momentum regions, and such discrepancies become smaller with the increase of the impact electron energies. It is suggested that the observed discrepancies are due to the distorted-wave effects in molecules, while appropriate theoretical calculations using distorted waves in molecules could not be achieved until now.

An investigation of valence shell orbital momentum profiles of difluoromethane by binary (e,2e) spectroscopy.

The electron binding energy spectra and momentum profiles of the valence orbitals of difluoromethane, also known as HFC32 (HFC-hydrofluorocarbon) (CH(2)F(2)), have been studied by using a high resolution (e,2e) electron momentum spectrometer, at an impact energy of 1200 eV plus the binding energy, and by using symmetric noncoplanar kinematics. The experimental momentum profiles of the outer valence orbitals and 4a(1) inner valence orbital are compared with the theoretical momentum distributions calculated using Hartree-Fock and density functional theory (DFT) methods with various basis sets. In general, the shapes of the experimental momentum distributions are well described by both the Hartree-Fock and DFT calculations when large and diffuse basis sets are used. However, the result also shows that it is hard to choose the different calculations for some orbitals, including the methods and the size of the basis sets employed. The pole strength of the ionization peak from the 4a(1) inner valence orbital is estimated.

The outer valance orbital electron densities of cyclopentane by binary (e,2e) spectroscopy.

The binding energy spectra and electron distributions in momentum space of the valence orbitals of cyclopentane (C(5)H(10)) are studied by Electron Momentum Spectroscopy (EMS) in a noncoplanar symmetric geometry. The impact energy was 1200 eV plus binding energy and energy resolution of the EMS spectrometer was 1.2 eV. The experimental momentum profiles of the outer valence orbitals are compared with the theoretical momentum distributions calculated using Hartree-Fock and density functional theory (DFT) methods. The shapes of the experimental momentum distributions are generally quite well described by both the Hartree-Fock and DFT calculations when the large and diffuse basis sets are used.

Increased severity of alcoholic liver injury in female rats: role of oxidative stress, endotoxin, and chemokines.

Alcoholic liver injury is more severe and rapidly developing in women than men. To evaluate the reason(s) for these gender-related differences, we determined whether pathogenic mechanisms important in alcoholic liver injury in male rats were further upregulated in female rats. Male and age-matched female rats (7/group) were fed ethanol and a diet containing fish oil for 4 wk by intragastric infusion. Dextrose isocalorically replaced ethanol in control rats. We analyzed liver histopathology, lipid peroxidation, cytochrome P-450 (CYP)2E1 activity, nonheme iron, endotoxin, nuclear factor-kappa B (NF-kappa B) activation, and mRNA levels of cyclooxygenase-1 (COX-1) and COX-2, tumor necrosis factor-alpha (TNF-alpha), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-2 (MIP-2). Alcohol-induced liver injury was more severe in female vs. male rats. Female rats had higher endotoxin, lipid peroxidation, and nonheme iron levels and increased NF-kappa B activation and upregulation of the chemokines MCP-1 and MIP-2. CYP2E1 activity and TNF-alpha and COX-2 levels were similar in male and female rats. Remarkably, female rats fed fish oil and dextrose also showed necrosis and inflammation. Our findings in ethanol-fed rats suggest that increased endotoxemia and lipid peroxidation in females stimulate NF-kappa B activation and chemokine production, enhancing liver injury. TNF-alpha and COX-2 upregulation are probably important in causing liver injury but do not explain gender-related differences.

Comorbid illness is an important determinant of health-related quality of life in patients with chronic hepatitis C.

OBJECTIVES: Chronic hepatitis C (CHC) patients selected for entry into treatment trials have been reported to have impaired health-related quality of life (HRQOL). However, these trials have an inherent selection bias, and HRQOL in CHC patients may have been underestimated because of the exclusion of patients with comorbid illness. The aim of this study was to assess HRQOL in an unselected group of CHC patients and to identify factors associated with impairment in HRQOL. METHODS: A total of 220 consecutive eligible CHC patients were enrolled from a hepatology clinic. HRQOL was assessed by the short form 36 (SF-36) and comorbid illnesses were assessed by an interview. RESULTS: CHC patients had significantly lower SF-36 scores in all subscales and in the summary scales when compared to those of the healthy general population in the United States (p < 0.001). Compared to CHC patients entering treatment trials, our patients had lower SF-36 scores on five subscales (p < 0.001). The presence of comorbid illness was the most important predictor of HRQOL in CHC patients. However, CHC alone resulted in significantly lower SF-36 scores in all subscales and summary scales (p < or = 0.003) compared to those of the healthy U.S. population. There was no correlation between SF-36 scores and history of i.v. drug use or dependence. alcohol dependence. and serum aminotransferase levels. CONCLUSIONS: We conclude that unselected CHC patients presenting for medical evaluation have a reduced HRQOL, which is lower than that reported for CHC patients entering treatment trials. CHC alone is associated with significant impairment in HRQOL, but the presence of comorbid illness leads to further diminution in HRQOL.

Outcome of liver transplantation for hepatitis B: report of a single center's experience.

Results of liver transplantation (LT) for hepatitis B have improved significantly with the use of hepatitis B immune globulin (HBIG) and/or lamivudine. The aim of this study is to review the long-term outcome of patients who underwent LT for hepatitis B. Records of 41 patients who underwent LT for hepatitis B and survived 3 months or longer post-LT were reviewed. Twenty patients were administered no immunoprophylaxis or short-term intramuscular HBIG, whereas 21 patients were administered high-dose intravenous (IV) HBIG. Median post-LT follow-up in these 2 groups was 76 months (range, 4 to 155 months) and 25 months (range, 4 to 68 months), respectively. Hepatitis B recurred in 15 (75%) and 4 patients (19%) who underwent LT in the pre-HBIG and post-HBIG eras, respectively. Cumulative rates of recurrent hepatitis B at 1 and 3 years post-LT in these 2 groups were 66% and 77% and 20% and 20%, respectively (P <.001). Recurrent hepatitis B in the post-HBIG era correlated with antibody to hepatitis B surface antigen titer less than 100 IU/L. Nine patients with recurrent hepatitis B were administered lamivudine for 13 to 49 months (median, 28 months); 6 patients continued to have stable or improved liver disease, whereas 3 patients developed virological breakthrough with slow deterioration of liver disease. Long-term IV HBIG is effective in preventing recurrent hepatitis B. The risk for recurrent hepatitis B is negligible after the first year post-LT. Among patients with no virological breakthrough, lamivudine can stabilize or improve liver disease for up to 4 years in patients with recurrent hepatitis B post-LT.

Protegrin-1 enhances bacterial killing in thermally injured skin.

OBJECTIVE: Septic complications and the emergence of drug-resistant microbes represent serious risks to patients. Recently, naturally occurring peptides have been discovered that possess potent and broad-spectrum antimicrobial activity. Protegrin-1 is particularly attractive for clinical use in human wounds because, unlike defensins, protegrin-1 retains broad antimicrobial and antifungal activity at physiologic salt concentration and in the presence of serum. The objective of this study was to examine the efficacy of protegrin-1 in killing multiple drug-resistant microbes isolated from human burn patients. DESIGN: For thein vitroexperiment, bilayer radial diffusion was performed comparing standard antibiotics with protegrin-1 on multiple-drug-resistant microbial organisms isolated from infected burn wounds. In vivo, rats received a 20% total body surface area partial-thickness burn by immersion in 60 degrees C water for 20 secs followed by wound seeding with 106 colony forming units of Silvadene-resistant Pseudomonas aeruginosa. SETTING: University of Michigan research laboratory. SUBJECTS: Adult, male Sprague-Dawley rats. INTERVENTIONS: Rats were randomized into three groups: those receiving synthetic protegrin-1, acetic acid (carrier), or gentamicin (positive control). Protegrin-1 was administered by topical application or intradermal injection. Wound tissues were harvested aseptically at different time points for quantitative bacterial counts. MEASUREMENTS AND MAIN RESULTS: In vivo and in vitro experiments revealed rapid and significant decreases in bacterial counts for protegrin-1-treated groups compared with controls. CONCLUSIONS: This study shows that protegrin-1 potentially may be used as an alternative or adjunct therapy to standard agents used to treat wound infections.

Feasibility of biolistic gene therapy in burns.

Skin is an especially attractive target for genetic manipulation because it is readily accessible and easily monitored for both the presence and the expression of inserted genes. This study was designed to assess the feasibility of particle mediated gene transfer to burned skin and to compare the transfection efficiency, anatomic distribution, and duration of transgene expression achievable in normal versus burned skin. Two days following scald injury of varying depths in 60 degrees C water (10 s: superficial partial; 20 s: deep partial; 40 s: full thickness) reporter gene (beta-galactosidase) constructs were delivered using a gene gun at various helium pressures (200-600 psi) to normal and burned skin. A time course study was performed to examine the kinetics of transgene expression. Animals received a superficial partial thickness burn and were sacrificed 12 h, 1, 3, 5, 7, 14, or 21 days after gene transfer. India Ink injection and immunohistochemistry were used to assess the depth of the scald injury. Transfection efficiency was measured in skin homogenates 24 h after gene transfer by morphometric and chemoluminescent assays. We found that the extent of tissue damage was directly related to the duration of heat source exposure. Reporter gene activity was significantly higher in superficial partial thickness burns compared to normal controls and gradually declined with increasing tissue injury. No activity was seen in the full thickness burn group. Beta-galactosidase activity reached a maximum level 12 h after gene transfer in both normal and superficial partial thickness burned skin with no levels seen after 5 days post-transfection. These findings indicate that particle-mediated gene transfer in thermally injured skin is feasible and may provide a means of introducing biologic agents into injured tissue capable of enhancing bacterial clearance and improving wound healing.

Lipopolysaccharide-binding protein accelerates and augments Escherichia coli phagocytosis by alveolar macrophages.

BACKGROUND: The first step in bacterial clearance by leukocytes is attachment and phagocytosis. Although lipopolysaccharide-binding protein (LBP) is best known for potentiating LPS-induced cytokine production through a CD14-dependent pathway, recent studies suggest that LBP plays a critical role in clearance of gram-negative bacteria and is essential for survival after bacterial challenge. We therefore sought to examine LBP's effect on Escherichia coli phagocytosis by alveolar macrophages (AMs) and to determine if this effect is mediated through CD14. MATERIALS AND METHODS: Phosphatidylinositol-specific phospholipase C (PIPLC)-treated and untreated rat AMs were incubated in the presence of increasing doses of recombinant LBP or negative control protein (choramphenicol acetyltransferase) prior to E. coli-FITC (Ec-F) BioParticle challenge. Phagocytosed bacteria were assayed by fluorescence measurement. A time course study was also performed. RESULTS: LBP potentiated phagocytosis of Ec-F BioParticles by AMs in a dose-dependent fashion. Kinetic studies showed that LBP augmented Ec-F phagocytosis by 76% at 30 min. Treatment of AMs with PIPLC to remove CD14 resulted in only a partial decrease in LBP-mediated enhancement of phagocytosis. CONCLUSION: These results clearly demonstrate that LBP plays an important role in enhancing Ec-F binding and phagocytosis in a time- and dose-dependent manner. This observed increase may not require the presence of CD14 as significant potentiation of phagocytosis still occurred after PIPLC treatment. We postulate that the LBP-mediated increase in Ec-F phagocytosis can occur in the absence of CD14 through the presence of another receptor.

Skin lipopolysaccharide-binding protein and IL-1beta production after thermal injury.

In response to a burn injury, skin can have an inflammatory response characterized by the production of inflammatory cytokines, recruitment of immune cells, containment of invading organisms, and clearance of noxious substances from the wound. Lipopolysaccharide-binding protein (LBP) is a molecule that is capable of coordinating all 4 functions; we previously found evidence that suggested that LBP is produced within surgical wounds. Because of the central role of LBP in the response to bacterial infection, as well as in the high rate of infection after burn injuries, we sought to determine whether a thermal injury could affect wound LBP production and thereby affect host responses against bacterial infection. Rats were given either a burn or a sham burn and were killed 24, 48, and 72 hours after the injuries. Wound specimens were assayed for bacterial counts and for the presence of LBP, messenger (m)RNA, and interleukin (IL)-1beta mRNA. Wound LBP mRNA was significantly upregulated at 24 hours in the group with burn injuries (P < .05; burn vs sham burn); this was followed by decreases at 48 and 72 hours. Immunohistochemistry showed LBP protein in the epidermis of animals with burns. Bacterial counts increased in the group with burn injuries (P < .05; burn vs sham burn) and continued to rise for 72 hours. IL-1beta mRNA levels were elevated at all time points in the group with burn injuries (P < .05). These results suggest an inverse correlation between burn wound LBP expression and bacterial wound counts. This failure to maintain local LBP production after severe thermal injury despite localized inflammation shown by high IL-1beta levels may predispose local wounds to bacterial invasion.

Mechanism of the alcohol cyclic pattern: role of the hypothalamic-pituitary-thyroid axis.

The cause of the cycle of urinary alcohol levels (UALs) in rats fed ethanol continually at a fixed rate is unknown. Rats were fed ethanol intragastrically at a constant dose for 2 mo, and daily body temperatures and UALs were recorded. Body temperature cycled inversely to UAL, suggesting that the rate of metabolism could be mechanistically involved in the rate of ethanol elimination during the cycle. To document this, whole body O(2) consumption rate was monitored daily during the cycle. The rate of O(2) consumption correlated positively with the change in body temperature and negatively with the change in UAL. Since the metabolic rate responds to changes in body temperature, thyroid hormone levels were measured during the UAL cycle. T(4) levels correlated positively with the O(2) consumption rate and negatively with the UALs. In a second experiment using propylthiouracil treatment, UALs did not cycle and a fall in body temperature failed to stimulate an increase in the rate of ethanol elimination. Consequently, rats died of overdose. Likewise, in a third experiment using rats with severed pituitary stalks, UALs failed to cycle and rats died of overdose. From these observations it was concluded that the UAL cycle depends on an intact hypothalamic-pituitary-thyroid axis response to the ethanol-induced drop in body temperature by increasing the rate of ethanol elimination.

Kupffer cell activation by lipopolysaccharide in rats: role for lipopolysaccharide binding protein and toll-like receptor 4.

Lipopolysaccharide (LPS) binding protein (LBP) is a key serum factor that mediates LPS activation of mononuclear cells. In the presence of LBP, 1/1,000 the concentration of LPS is sufficient to activate peripheral blood monocytes. Previous studies with Kupffer cells have shown a variable effect of serum on LPS activation of these cells and led to the conclusion that, unlike extrahepatic mononuclear cells, Kupffer cells do not respond to LPS in an LBP-dependent fashion. Because there are multiple components in serum other than LBP that might affect LPS activation, these reports with serum are difficult to interpret. To investigate the specific role of LBP in LPS activation of Kupffer cells, we produced a functional recombinant rat LBP using a baculovirus expression system, which we used to selectively examine the role of LBP's on Kupffer-cell function. Isolated Kupffer cells exposed to increasing concentrations of LPS (0, 1, 10 ng/mL) showed a dose-dependent increase in TNF-alpha production, which was augmented and accelerated by the presence of LBP. The effects of LBP on Kupffer cell activation by LPS are dependent on a functional Toll-like receptor 4 (Tlr 4) because Kupffer cells from C3H/HeJ mice failed to respond to LPS in the presence of LBP. LBP plays an important role in mediating Kupffer cell activation by LPS, and these effects are dependent on the presence of functioning Tlr 4.