[A combined method of proximalization of the patella tendon and double boundle reconstruction of medial patellofemoral ligament with adductor magnus tendon autograft for the treatment of habitual dislocation of patella in adolescent].

Objective: To present the technique and clinic effect of the combined method of proximalization of the patella and double boundle reconstruction of medial patellofemoral ligament (MPFL) with adductor magnus tendon autograft for the treatment of habitual dislocation of patella (HDP) in adolescent. Methods: From August 2016 to August 2018, a total of 33 consecutive skeletally immature adolescent patients (37 knees) with HDP were surgically treated in Beijing Jishuitan Hospital and were retrospectively reviewed. Among them, 10 patients (12 knees) with severe quadriceps contracture underwent the index comprehensive procedure. There were 4 males and 6 females with a mean age of (12.1±1.4) years (10 to 14 years old) at the operation. Before surgery and at the final follow-up, subjective symptoms were scored by Lysholm knee score, physical examination and radiological assessment were performed. Results: of patellar tracking were assessed by congruence angle and lateral patellofemoral angle. Results Patients were followed up for an average period of 23 months (12-36 months). No infection, patella redislocation were observed in all cases. Lysholm scores improved from 77±9 before surgery to 96±6 at the final follow-up (t= -23.155, P<0.05). There was a statistically significant improvement in the congruence angle, from 72.4°±17.2° preoperatively to -7.5°±4.8° at the final follow-up (t=21.392, P<0.01) and in the lateral patellofemoral angle, from -64.6°±9.4° preoperatively to 6.5°±3.7° at the final follow-up (t=-22.874,P<0.01). Conclusion: In this short term study, the novel comprehensive procedure, including proximalization of the patella and double boundle reconstruction of MPFL with adductor magnus tendon autograft, treats HDP effectively in skeletally immature adolescent patients with severe quadriceps contracture.

[Single portal arthroscopic-assisted reduction technique in the treatment of irreducible developmental dislocation of the hip in infants and toddlers].

Objective: To investigate the efficacy of single-portal arthroscopic-assisted reduction technique in the developmental dislocation of the hip in infants. Methods: From January 2014 to December 2016, 12 dislocated hips in 12 children with a median age of 14 months (10 to 20 months) were treated with single-portal arthroscopic-assisted reduction technique. The indication for intervention was failure of closed reduction after bilateral adductor and unilateral iliopsoas release under anesthesia. Adductor and iliopsoas tendon were released routinely through a medial approach. Through the same medial approach a single-portal arthroscopic-assisted reduction technique was selected, with a medial sub-adductor portal for both 4.0 mm cannulated system with a 30° arthroscope and the instruments. After assessing of the intraarticular structures, the hypertrophic ligamentum teres and acetabular pulvinar were resected, and transverse acetabular ligament (TAL) was incised and a limited release of the capsule was performed prior to reduction of the hip. Arthrography was performed after reduction in all children. Safe zone angle and the medialization ratio on the arthrogram were compared pre and post arthroscopic reduction. Acetabular index were compared at two time points: before operation and at the latest follow-up with paired t test. Results: All hips were reduced with single-portal arthroscopic procedures. The reduction was confirmed on arthrography. With a median follow-up of 26 months (18 to 36 months), all 12 hips remained stable. Safe zone angle increased from 18.5°±3.8° to 61.9°±6.5° immediately after arthroscopic reduction(t=-28.944, P<0.01); and the medialization ratio on the arthrogram increased from 62%±20% to 104%±16% immediately after arthroscopic reduction (t=-3.519, P<0.05). The mean acetabular index decreased from 40.6°±5.0° to 29.4°±5.0° at the latest follow-up (t=5.463, P<0.01). However, Kalamchi-MacEwen type Ⅰ avascular necrosis was developed only in 1 case, and residual dysplasia was observed in 2 hips. Conclusions: Single-portal arthroscopic-assisted reduction technique is a safe and effective treatment for developmental dislocation of the hip in infants and toddlers.

Nicotinamide N-methyltransferase as a potential marker for cancer.

Cancer is the main leading cause of death in the world, although it has been made noteworthy advances in cancer research in the past decades. Early detection of cancer is extremely important in improving the chances of successful therapy. Thus, it is urgently needed to make further efforts to explore novel tumor markers for treatment. Nicotinamide N-methyltransferase (NNMT) is a cytosolic enzyme which catalyzes the N-methylation of nicotinamide to form 1-methylnicotinamide (1-MNA), and plays an important role in controlling the intracellular concentration of nicotinamide. Nicotinamide, the precursor to NAD+, is an important cofactor that associates cellular redox states with energy metabolism. Growing evidence shows that NNMT protein levels are elevated in a variety of human cancers, and increased NNMT expression has been linked to tumor aggressiveness. This paper presents a review for the role of NNMT expressed in a series of human cancers and the regulating mechanism involved, and offers its potential value of NNMT in cancer detection and treatment.

[Clinical application of 125I radioactive seeds brachytherapy in the treatment of the pediatric soft tissue sarcoma in head and neck].

Objective: To investigate the clinical application and preliminary results of 125I radioactive seeds brachytherapy in the comprehensive treatment of the pediatric soft tissue sarcoma in head and neck. Methods: A total of 24 pediatric patients with soft tissue sarcoma in head and neck were treated at Peking University School of Stomatology from April 2012 to July 2015. The data was collected and analyzed through statistical methods, which included the pathological type, gender, age, tumor location, volume, treatment and the clinical results after the application of 125I radioactive seeds brachytherapy. Results: Among the 24 patients, there were 18 patients of rhabdomyosarcoma, 2 Ewing's sarcoma, 2 fibrosarcoma, 1 synovial sarcoma and 1 malignant fibrous histiocytoma. After a follow-up of 9-48 months, 1 case of temporal rhabdomyosarcoma was observed to have a progression to the the lateral cranial base at the time of 12 months, 2 cases realized local control and systemic progression, the rest were completely relieved, and there was no recurrence during the follow-up period.The Kaplan-Meier survival analysis showed that the 1-year and 3-year local control rate were both 94.1%, the 1-year and 3-year event-free survival rate were 87.4% and 77.7%. Conclusion: The preliminary results indicate that 125I radioactive seeds brachytherapy play a very important role in the improvement of the local control rate and survival quality of the pediatric soft tissue sarcoma patient in head and neck, and it's a minimally invasive treatment with the advantage of accuracy andsmall side effects.

[Study on the impact of labro-chondral complex on development of the acetabulum after close reduction in developmental dysplasia of the hip].

Objective: To explore the influence of the labro-chondral complex (LLC) on the development of the acetabulum after close reduction in developmental dysplasia of the hip (DDH). Methods: Sixty-one cases (72 hips) with DDH presented in Beijing Jishuitan Hospital were reviewed, all the patients were treated by closed reduction, arthrogram and Spica casting from March 2010 to December 2013. The anterior-posterior pelvic radiography was performed to evaluate the morphology of the labro-chondral complex and reduction of the hip. The cases were divided into Ⅰ, Ⅱ, Ⅲ, Ⅳ four groups according to the shape of the LLC initially, and when performed the secondary Spica cast after 3 months, these cases were divided into 0-0.4, 0.4-0.6, and >0.6, three groups based on the height difference ratio (HDR) of the LLC. The relationship between the shape and HDR of the LLC was analyzed. The AI and CE angle were used to evaluate development of the hip during the latest follow up. The impact of the shape and HDR of the LLC on the development of the acetabulum was explored as well. Results: The HDR was the least in the type Ⅰ hips, all cases were less than 0.6, the AI in this group was significantly lower than the others(24.33°±3.12°), and the CE angle was significantly higher in the type Ⅰ hips(15.22°±3.11°) during the latest follow up. The CE angle was significantly different among the three groups of HDR. The HDR was lower, the CE angle was higher. The AI in 0-0.4 group was significantly lower than the others(14.24°±3.10°). Conclusion: The shape of the LLC is helpful to judge development of the acetabulum when closed reduction was performed in DDH. And the HDR in the secondary cast change could be used as a sensitive index to predict development of the hip.

Nitric oxide activates intradomain disulfide bond formation in the kinase loop of Akt1/PKBα after burn injury.

Severe burn injury is an acute inflammatory state with massive alterations in gene expression and levels of growth factors, cytokines and free radicals. During the catabolic processes, changes in insulin sensitivity and skeletal muscle wasting (unintended loss of 5-15% of lean body mass) are observed clinically. Here, we reveal a novel molecular mechanism of Akt1/protein kinase Bα (Akt1/PKBα) regulated via cross-talking between dephosphorylation of Thr308 and S-nitrosylation of Cys296 post severe burn injury, which were characterized using nano-LC interfaced with tandem quadrupole time-of-fight mass spectrometry (Q-TOF)micro tandem mass spectrometry in both in vitro and in vivo studies. For the in vitro studies, Akt1/PKBα was S-nitrosylated with S-nitrosoglutathione and derivatized by three methods. The derivatives were isolated by SDS-PAGE, trypsinized and analyzed by the tandem MS. For the in vivo studies, Akt1/PKBα in muscle lysates from burned rats was immunoprecipitated, derivatized with HPDP-Biotin and analyzed as above. The studies demonstrated that the NO free radical reacts with the free thiol of Cys296 to produce a Cys296-SNO intermediate which accelerates interaction with Cys310 to form Cys296-Cys310 in the kinase loop. MS/MS sequence analysis indicated that the dipeptide, linked via Cys296-Cys310, underwent dephosphorylation at Thr308. These effects were not observed in lysates from sham animals. As a result of this dual effect of burn injury, the loose conformation that is slightly stabilized by the Lys297-Thr308 salt bridge may be replaced by a more rigid structure which may block substrate access. Together with the findings of our previous report concerning mild IRS-1 integrity changes post burn, it is reasonable to conclude that the impaired Akt1/PKBα has a major impact on FOXO3 subcellular distribution and activities.

Combining breast and cervical screening in an attempt to increase cervical screening uptake. An intervention study in a South African context.

During 2008-2009, only 4% of women targeted for cervical screening were screened in Tshwane, South Africa. The purpose of the study was to determine whether cervical screening uptake could be improved when breast and cervical screening are combined. An intervention research design was used. The intervention was assessed in terms of two outcomes, namely cervical screening uptake and the findings of the screening. The study was conducted in a resource poor environment in Tshwane. Convenience sampling was used to recruit the sample (n = 299) and a baseline survey was conducted before delivering the intervention. Only 14% of the sample (n = 299) reported having been screened for cervical cancer previously. The total sample (n = 299) were willing to have a clinical breast examination; however, only 65.4% of those eligible for cervical screening (n = 283) used the opportunity to be screened. The majority of the sample screened (n = 185) using acetic acid for visual inspection (VIA) were VIA negative; 12.4% were VIA positive and 4.4% were VIA positive, invasive cancer; the screening of 8.7% failed. Despite women's lack of knowledge of cervical cancer and the screening thereof, combining cervical screening and breast screening lead to an increase in cervical screening uptake.

SILAM for quantitative proteomics of liver Akt1/PKBα after burn injury.

Akt1/protein kinase Bα (Akt1/PKBα) is a downstream mediator of the insulin signaling system. In this study we explored mechanism(s) for its role in burn injury. Akt1/PKBα in liver extracts from mice with burn injury fed with (2H7)-L-Leu was immunoprecipitated and isolated with SDS-PAGE. Two tryptic peptides, one in the kinase loop and a control peptide just outside of the loop were sequenced via nano-LC interfaced with quadruple time-of-flight tandem mass spectrometry (Q-TOF tandem MS). Their relative isotopologue abundances were determined by stable isotope labeling by amino acids in mammalians (SILAM). Relative quantifications based on paired heavy/light peptides were obtained in 3 steps. The first step included homogenization of mixtures of equal amounts of tissue from burned and sham-treated animals (i.e., isotope dilution) and acquisition of uncorrected data based on parent monoisotopic MS ion ratios. The second step included determination of isotopic enrichment of the kinase from burned mice on Day 7 and the third step enrichment correction of partially labeled heavy and light monoisotopic MS ion ratios for relative quantification of bioactivity (loop peptide) and expression level (control peptide). Protein synthesis and enrichment after injury were found to be dependent on tissue and turnover of individual proteins. Three heavy and light monoisotopic ion ratios for albumin peptides from burned mice indicated ~55% enrichment and ~16.7-fold downregulation. In contract, serum amyloid P had ~66% enrichment and was significantly upregulated. Akt1/PKBα had ~56% enrichment and kinase level in response to the burn injury was upregulated compared with the control peptide. However, kinase bioactivity, represented by the Cys296 peptide, was significantly reduced. Overall, we demonstrated that i) quantitative proteomics can be performed without completely labeled mice; ii) measurement of enrichment of acyl-tRNAs is unnecessary and iii) Cys296 plays an important role in kinase activity after burn injury.

Human papillomavirus in esophageal squamous cell carcinoma of the high-risk Kazakh ethnic group in Xinjiang, China.

AIMS: To investigate human papillomavirus (HPV) genotype-specific prevalence in the high-risk Kazakh ethnic group with esophageal squamous cell carcinoma (ESCC). METHODS: Sixty-seven Kazakh patients with primary ESCC were studied. From each patient, two tissue samples were collected: one sample of the tumor and one sample of normal esophageal tissue from an area away from the tumor. Tissues were analyzed by INNO-LiPA HPV Genotyping test v2 assay allowing the detection of at least 24 different HPV genotypes. RESULTS: Twenty cancer patients (30%) had HPV DNA detected in collected specimens. Interestingly, 14 patients (21%) had HPV only in the tumor and six (9%) had HPV only in the normal esophageal tissue. Overall, HPV16 was the viral type most frequently detected being present in eight out of 20 positive cases (40%). No correlation between the presence of HPVs and the gender or ESCC grade was observed. CONCLUSION: If the causative factors of esophageal carcinogenesis remain to be firmly established in the Kazakh population, HPV found in 30% of patients might play a role in the etiology of esophageal SCC.

Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus

The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group) infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 103) were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1) and albendazole (50 mg kg-1 day-1), separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1). Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01), but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1 percent) in the group receiving the combined treatments (18.3 ± 4.6 mg). IL-4 and total IgE were decreased (939 ± 447 pg/mL and 2.03 ± 0.42 IU/mL, respectively) in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 ± 619 pg/mL and 3.31 ± 0.37 IU/mL; P < 0.01). These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus.

The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group) infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 10(3)) were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1) and albendazole (50 mg kg-1 day-1), separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1). Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01), but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1%) in the group receiving the combined treatments (18.3 +/- 4.6 mg). IL-4 and total IgE were decreased (939 +/- 447 pg/mL and 2.03 +/- 0.42 IU/mL, respectively) in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 +/- 619 pg/mL and 3.31 +/- 0.37 IU/mL; P < 0.01). These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

Synthesis of a macromolecular camptothecin conjugate with dual phase drug release.

A water soluble macromolecular conjugate of camptothecin (CPT) with a new, dual phase hydrolytic drug release mechanism was prepared on the basis of a 60 kDa biodegradable hydrophilic "stealth" polyacetal, poly(1-hydroxymethylethylene hydroxy-methyl formal). Succinamido-glycinate was used as a prodrug releasing group. A model preparation with 7.5% CPT content w/w was water soluble. The lipophilic camptothecin prodrug, camptothecin-(O20)-succinimidoglycinate, was released from the conjugate with t(1/2) = 2.2 +/- 0.1 h in rodent plasma. The blood clearance in a rodent model as measured by CPT was release limited, t(1/2) = 2.1 +/- 0.2 h, while the conjugate half-life was 14.2 +/- 1.7 h. In a xenograft tumor model, the conjugate demonstrated higher antineoplastic efficacy than CPT at a less than equitoxic dose. This improved therapeutic window is in line with the modified drug pharmacokinetics and with camptothecin release in a stabilized lipophilic prodrug form. Regulation of prodrug release and hydrolysis rates through linker structure modification will open the way to further improve both pharmacokinetics and pharmacodynamics.

Treatment with the snail peptide CGX-1007 reduces DNA damage and alters gene expression of c-fos and bcl-2 following focal ischemic brain injury in rats.

Delayed cell death following ischemic brain injury has been linked to alterations in gene expression. In this study we have evaluated the upregulation of several genes associated with delayed cell death (c-fos, bax, and bcl-2) during the initial 24 h of transient middle cerebral artery occlusion (MCAo) in the rat and the effects of postinjury treatment with the NR2B subunit specific NMDA receptor antagonist CGX-1007 (Conantokin-G, Con-G). C-fos mRNA levels peaked at 1 h postinjury in both cortical and subcortical ischemic brain regions (30-fold increase), remained elevated at 4 h and returned to within normal, preinjury levels 24 h postinjury. The increase in mRNA levels correlated to increased protein expression in the entire ipsilateral hemisphere at 1 h. Regions of necrosis at 4 h were void of C-Fos immunoreactivity with continued upregulation in surrounding regions. At 24 h, loss of C-Fos staining was observed in the injured hemisphere except for sustained increases along the border of the infarct and in the cingulate cortex of vehicle treated rats. CGX-1007 treatment reduced c-fos expression throughout the infarct region by up to 50%. No significant differences were measured in either bcl-2 or bax mRNA expression between treatment groups. However, at 24 h postinjury CGX-1007 treatment was associated with an increase in Bcl-2 immunoreactivity that correlated to a reduction in DNA fragmentation. In conclusion, CGX-1007 effectively attenuated gene expression associated with delayed cell death as related to a neuroprotective relief of cerebral ischemia.

Cysteine metabolism and whole blood glutathione synthesis in septic pediatric patients.

OBJECTIVE: To investigate whole body in vivo cysteine kinetics and its relationship to whole blood glutathione (GSH) synthesis rates in septic, critically ill pediatric patients and controls. DESIGN: Prospective cohort study. SETTING: Multidisciplinary intensive care unit and pediatric inpatient units at a children's hospital. PATIENTS: Ten septic pediatric patients and ten controls (children admitted to the hospital for elective surgery). INTERVENTIONS: Septic patients (age, 31 months to 17 yrs) and controls (age, 24 months to 21 yrs) received a 6-hr primed, constant, intravenous tracer infusion of l-[1-13C]cysteine. Blood samples were obtained to determine isotopic enrichment of plasma cysteine and whole blood [1-13C]cysteinyl-glutathione by gas-chromatography mass spectrometric techniques. The plasma flux and oxidation rate of cysteine and the fractional and absolute synthesis rates of GSH were determined. Septic patients received variable protein and energy intake, as per routine clinical management, and controls were studied in the early postabsorptive state. MEASUREMENTS AND MAIN RESULTS: Plasma cysteine fluxes were increased in the septic patients when compared with the controls (68.2 +/- 17.5 [sd] vs. 48.7 +/- 8.8 micromol x kg(-1) x hr(-1); p <.01), and the fraction of plasma cysteine flux associated with oxidative disposal was similar among the groups. The absolute rates of GSH synthesis in whole blood were decreased (p <.01) in the septic patients (368 +/- 156 vs. 909 +/- 272 micromol x L(-1) x day(-1)). The concentration of whole blood GSH also was decreased in the septic group (665.4 +/- 194 vs. 1059 +/- 334 microM; p <.01) CONCLUSIONS: Whole blood glutathione synthesis rates are decreased, by about 60%, in critically ill septic children receiving limited nutritional support. Plasma cysteine fluxes and concentration of cysteine were increased in the septic patients, suggesting a hypermetabolic state with increased protein breakdown. The mechanisms whereby GSH synthesis rates are decreased in these patients are probably multifactorial, presumably involving an inflammatory response in the presence of limited nutritional support. The role of nutritional modulation and the use of cysteine prodrugs in maintaining GSH concentration and synthesis remain to be established.

Blood glutathione synthesis rates in healthy adults receiving a sulfur amino acid-free diet.

The availability of cysteine is thought to be the rate limiting factor for synthesis of the tripeptide glutathione (GSH), based on studies in rodents. GSH status is compromised in various disease states and by certain medications leading to increased morbidity and poor survival. To determine the possible importance of dietary cyst(e)ine availability for whole blood glutathione synthesis in humans, we developed a convenient mass spectrometric method for measurement of the isotopic enrichment of intact GSH and then applied it in a controlled metabolic study. Seven healthy male subjects received during two separate 10-day periods an L-amino acid based diet supplying an adequate amino acid intake or a sulfur amino acid (SAA) (methionine and cysteine) free mixture (SAA-free). On day 10, L-[1-(13)C]cysteine was given as a primed, constant i.v. infusion (3 micromol x kg(-1) x h(-1)) for 6 h, and incorporation of label into whole blood GSH determined by GC/MS selected ion monitoring. The fractional synthesis rate (mean +/- SD; day(-1)) of whole blood GSH was 0.65 +/- 0.13 for the adequate diet and 0.49 +/- 0.13 for the SAA-free diet (P < 0.01). Whole blood GSH was 1,142 +/- 243 and 1,216 +/- 162 microM for the adequate and SAA-free periods (P > 0.05), and the absolute rate of GSH synthesis was 747 +/- 216 and 579 +/- 135 micromol x liter(-1) x day(-1), respectively (P < 0.05). Thus, a restricted dietary supply of SAA slows the rate of whole blood GSH synthesis and diminishes turnover, with maintenance of the GSH concentration in healthy subjects.

Oxoproline kinetics and oxoproline urinary excretion during glycine- or sulfur amino acid-free diets in humans.

L-5-oxoproline (L-5-OP) is an intermediate in glutathione synthesis, possibly limited by cysteine availability. Urinary 5-OP excretion has been proposed as a measure of glycine availability. We investigated whether 5 days of dietary sulfur amino acid (SAA-free) or glycine (Gly-free) restriction affects plasma kinetics of 5-OP and urinary excretion of L- and D-5-OP in 6 healthy men. On day 6, L-5-[1-(13)C]oxoproline and [3,3-(2)H(2)]cysteine were infused intravenously for 8 h (3 h fast/5 h fed). In a control study (adequate amino acid mixture), plasma oxoproline fluxes were 37.8 +/- 13.8 (SD) and 38.4 +/- 14.8 micromol x kg(-1) x h(-1); oxidation accounted for 85% of flux. Cysteine flux was 47.9 +/- 8.5 and 43.2 +/- 8.5 micromol x kg(-1) x h(-1) for fast and fed phases, respectively. Urinary excretion of L- and D-5-OP was 70 +/- 34 and 31.1 +/- 13.3 micromol/mmol creatinine, respectively, during days 3-5, and 46.4 +/- 13.9 and 22.4 +/- 8.3 micromol/mmol over the 8-h tracer study. The 5-OP flux for the Gly-free diet was higher (P = 0. 018) and tended to be higher for the SAA-free diet (P = 0.057) when compared with the control diet. Oxidation rates were higher on the Gly-free (P = 0.005) and SAA-free (P = 0.03) diets. Cysteine fluxes were lower on the the Gly-free (P = 0.01) and the SAA-free diets (P = 0.001) compared with the control diet. Rates of L-5-OP excretion were unchanged by withdrawal of SAA or Gly for 5 days but increased on day 6 (P = 0.005 and P = 0.019, respectively). Thus acute changes in the dietary availability of SAA and Gly alter oxoproline kinetics and urinary 5-OP excretion.

[Detection of EBV-DNA, EBNA2 and LMP-1 in tumor tissues of han and uygur patients with nasopharyngeal carcinoma in Xinjiang].

OBJECTIVE: To investigate the possible differences of human nasopharyngeal carcinogenesis between Han and Uygur patients in xinjiang. METHOD: Detection of EBV-DNA, EBNA2 and LMP-1 in tumor tissues of 73 patients (Han 41, uygur 32) with nasopharyngeal carcinoma (NPC) were performed by PCR and immunohistochemistry. RESULT: Tho positive rate of EBV-DNA, EBNA2 expression was 48.2%(20/41), 43.9% (18/41) in Han and 59.3%(19/32), 43.7%(14/32) in Uygur respectively, no significant difference was found (P > 0.05). Expression of LMP-1 in NPC, in both positive and negative EBV-DNA specimens, was higher in Uygur (78.9%) than that in Han (40%) (P < 0.05), CONCLUSION: The EBV infection may involve in nasopharyngeal carcinogenesis in xinjiang; the LMP-1 of EBV could be a more critical factor in malignant transformation of normal nasopharyngeal epithelium in uygur people than in han Chinese.

Plasma L-5-oxoproline carbon and nitrogen kinetics in healthy young adults.

L-5-oxoproline (OP), an intermediate of the gamma-glutamyl cycle of glutathione synthesis and degradation, may serve as a probe for the state of glutathione kinetics. We explored the whole-body carbon and nitrogen kinetics of OP in five male healthy subjects (75.2 kg; 181 cm; 26 y) after a 5-d adaptation to an adequate L-amino acid-based diet (160 mg N x kg(-1) x d(-1); 188 kJ x kg(-1) x d(-1)), using a crossover design. On day 6 of the diet period, we carried out an 8-h tracer protocol (3 h fast; 5 h fed; 2/3 of daily nitrogen intake) with intravenous infusion of L-[1-(13)C]oxoproline and L-[3, 3-(2)H]cysteine or, in randomized order, on the second occasion, L-[(15)N]oxoproline and L-[3,3-(2)H]cysteine. Plasma OP was isolated by cation exchange and after addition of internal standards (DL-[(2)H(3)]-5-oxoproline; L-[(15)N, U-(13)C(5)]-5-oxoproline; DL-[(2)H(3)]-glutamic acid) derivatized to form TBDMS esters and measured by gas chromatography/mass spectrometry. Plasma OP concentration did not differ between fed and fasted state (fast: 59. 4 +/- 8.3; fed 59.2 +/- 8.9 nmol/mL). (13)C- and (15)N OP flux during the fasted and fed state were 19 +/- 3.6, 21.2 +/- 3.2, and 22.6 +/- 3.9, 25.8 +/- 4.3 micromol x kg(-1) x 30 min(-1), respectively. OP oxidation was 15.6 +/- 3.6 and 17.9 +/- 3.5 micromol x kg(-1) x 30 min(-1), in fasting and feeding, respectively, (P < 0.05). More than 80% of the plasma flux was oxidized. These findings are compared with the published literature on GSH turnover in plasma of human subjects and underscore the need to define more completely the dynamic aspects of glutathione metabolism and of the intermediates of the gamma-glutamyl cycle.

Targeted antisense modulation of inflammatory cytokine receptors.

Antisense technology is potentially a powerful means by which to selectively control gene expression. We have used antisense oligonucleotides to modulate the response of the hepatoma cell line, HepG2, to the inflammatory cytokine, IL-6, by inhibiting the expression of its multifunctional signal transducer, gp130. HepG2 cells respond to IL-6 by upregulating acute phase proteins, such as haptoglobin, by five- to tenfold. Gp130 is central to this response, as the upregulation of haptoglobin is almost completely blocked by the addition of high concentrations ( approximately 100 microg/ml) of a monoclonal antibody to gp 130. Antisense oligodeoxynucleotides complementary to the mRNA encoding gp 130 inhibited the upregulation of haptoglobin by IL-6-stimulated HepG2 cells by about 50%. However, a nonsense sequence also inhibited haptoglobin secretion by about 20%. To improve the specificity and efficiency of action, we targeted the antisense oligonucleotides to HepG2 cells using a conjugate of asialoglycoprotein-poly-L-lysine. The targeted antisense reduced the binding of IL-6 to HepG2 cells, virtually eliminating high affinity binding. In addition, it inhibited haptoglobin upregulation by over 70%. Furthermore, the dose of targeted antisense required for biological effect was reduced by about an order of magnitude as compared with unconjugated antisense. These results demonstrate the potential of antisense oligonucleotides as a means to control the acute phase response as well as the need for a greater understanding of the mechanism and dynamics of antisense molecules as they are developed toward therapeutic application.