[The surgical effect on obstructive sleep apnea with laryngopharyngeal reflux].

Objective:To explore the effect of surgical treatment for OSA with laryngopharyngeal reflux（LPR）. Method:A retrospective analysis was made in 42 patients diagnosed as moderate to severe OSA with LPR and treated with modified-coblation assisted Uvulopalatopharyngoplasty（M-CAUP）. The results of PSG, reflux symptom index（RSI）, reflux finding score（RFS） and 24-hour esophageal pH monitoring before and after operation were compared. Result:The AHI after operation was significantly lower than that before operation（P<0.05）, and the average oxygen saturation and minimum blood pressure saturation were increased（P<0.05）. The total scores of RSI and RFS after operation were lower than those before operation（P<0.05）. The percentage of time of pH<4.0 in esophagus 24 hours after operation was lower than that before operation（P<0.05）. Conclusion:For patients with moderate to severe OSA combined with LPR with oropharyngeal obstruction, surgical treatment can not only reduce airway stenosis and obstruction, but also improve the symptoms and signs of LPR.

[Effect of different course of omeprazole in the treatment of laryngopharyngeal reflux].

Objective:To compare the efficacy and safety of omeprazole in the treatment of laryngopharyngeal reflux. Method:One hundred and sixty patients with laryngopharyngeal reflux in our hospital from March 2015 to January 2017 (diagnosis treatment for 8 weeks) were selected and randomly divided into group A, B, C, D, with 40 cases in each group. Each group of patients were treated with omeprazole enteric-coated capsules (20mg, Bid) for 1, 2, 3 and 4 months, respectively. The reflux symptom index(RSI), reflux finding score(RFS) score, and 24h intraesophageal pH-metry , pepsin detection were compared between the four groups before treatment, at the end of treatment, at 1 month and 2 months after treatment. The efficacy and side effects were evaluated.Result:With the increase of treatment course, patient's RSI and RFS score, 24h pH-metry <4.0 percentage, pepsin positive rate decreased gradually, and there was a significant difference between before and after treatment. After treatment, there was a statistical significance in group B and group A, group C and group B. There was no significant difference between group D and group C. After the end of treatment and up to 2 months of follow-up, the RSI and RFS scores gradually increased and 24h Ph-metry <4.0 percentage, pepsin positive rate increased in group A and B,and those were not significantly different in group C and group D. With the increase of treatment duration, the effective rate of treatment increased, which was 67.5%, 80.0%, 90.0% and 95.0%, respectively. The difference was statistically significant, but the adverse effects did not increase (P>0.05).Conclusion:Omeprazole is safe and effective in the treatment of laryngopharyngeal reflux. The symptoms and signs of the patients are obviously improved during the 5 months of the course, and the condition is relatively stable and the recurrence rate is low.

[Application of reflux symptom index in diagnosis of allergic patients with laryngopharyngeal reflux].

Objective:To explore the diagnostic value and optimal diagnostic threshold of reflux symptom index(RSI) in allergic patients with laryngopharyngeal reflux. Method: All the adult allergic patients with respiratory tract symptoms completed the RSI with the consent of the patients. A total of 150 patients with RSI＞13 were screened out. The LPR patients were confirmed by 24 h pH-metry of pharyngo-laryngoesophageal (allergic group). The same number of non allergic laryngeal reflux patients were selected as control group (non allergic group). The ratio of RSI＞13 to LPR was calculated. The total score of RSI and each score were compared between the allergic group and the non allergic group. ROC curve was used to analyze the highly suspicious RSI score thresholds for patients with reflux laryngitis. Result:Only 53(35.33%) of 150 patients with RSI＞13 were diagnosed as LPR after 24 h pH-metry test in allergic patients. Among the 9 symptoms of RSI, the scores of 6 symptoms in allergic group were significantly higher than those in non allergic group(P<0.05). The average RSI score of allergic group was 23.57±3.17. The average RSI score of non allergic group was 17.57±2.64. The total score of allergic group was higher than that of non allergic group(P<0.05). According to the RSI score ROC curve of allergic group, the area under 95% confidence interval curve was 0.815. RSI had certain diagnostic accuracy for allergic patients with LPR. The best critical value of RSI score for allergic patients with LPR was 18, the sensitivity was 94.3%, and the specificity was 56.7%. Conclusion:RSI can be used to screen allergic patients with LPR, diagnostic score threshold RSI＞18 points, has a certain diagnostic accuracy.

[A preliminary study on the relationship between laryngopharyngeal reflux and chronic rhinosinusitis].

Objective:To study the relationship between laryngopharyngeal reflux and chronic rhinosinusitis. Method:A total of 46 patients were enrolled in this study including 25 cases with chronic rhinosinusitis with nasal polyps, 10 cases with chronic rhinosinusitis without nasal polyps and 11 cases underwent surgery due to abnormal nasal anatomy such as nasal septum deviation, bubble in the turbinate, etc. as control group. The expression of pepsin was detected using immunohistochemistry in three groups. The intensity of pepsin expression and CT score of sinus, blood eosinophils percentage, blood neutrophils percentage, blood basohils percentage, blood mononuclear percentage, blood lymphocytes percentage were analyzed. Result:There were 8 strong positive cases (32%, 8/25), positive in 8 cases (32%, 8/25), 2 weakly positive cases (8%,2/25), 7 negative cases (28%, 7/25) in chronic rhinosinusitis with nasal polyps group. In the chronic rhinosinusitis without nasal polyps group, the expression of pepsin was strong positive in 4 cases (40%, 4/10), positive in 3 cases (30%, 3/10), weakly positive in 1 cases (10%, 1/10), negative in 2 cases (20%, 2/10). There were no strong positive expression in the control group, positive in 2 cases (18.2%, 2/11), weakly positive in 3 cases (27.3%, 3/11), negative in 6 cases (54.5%, 6/11), chronic rhinosinusitis with nasal polyps group and chronic rhinosinusitis without nasal polyps group higher than the control group (P<0.05). There was no significant difference in pepsin expression between chronic rhinosinusitis with nasal polyps group and without group (P=0.617). Spearman correlation analysis indicated that the intensity of pepsin was positively correlated with the score of Lund-Markay (r=0.349,P=0.017), there was no correlation with the percentage of various inflammatory cells. Conclusion:The positive expression intensity of pepsin in chronic rhinosinusitis without nasal polyps and chronic rhinosinusitis with nasal polyps is significantly higher than that in normal control group, suggested that there is a correlation between laryngopharyngeal reflux and chronic rhinosinusitis. Laryngopharyngeal reflux is positively correlated with the severity of nasal polyps. Chronic nasal inflammation caused by laryngopharyngeal reflux is not mediated by a certain kind of inflammatory cells.

Antagonistic effect of orphanin FQ on opioid analgesia in rat.

AIM: To study the effect of orphanin FQ (OFQ), a newly discovered heptadecapeptide, on nociception and opioid analgesia. METHODS: The intracerebroventricular (i.c.v.) and intrathecal (i.t.h.) injections were used to give the drugs. The tail-flick model of rats were used to test the pain threshold. RESULTS: OFQ (i.c.v. or i.t.h.) 0.1 microgram had no effect on nociception but 0.5-10 micrograms induces hyper-reaction of rat to noxious electric stimulus; the decapeptide (OFQ1-10 i.c.v.), a fragment of the OFQ, did not affect the pain reaction of rats. Fentanyl (1 microgram, i.c.v. or i.t.h.), a selective mu-receptor agonist, DSLET (5 micrograms, i.c.v. or i.t.h.), a selective delta-receptor agonist, or U50488H (1 microgram, i.t.h.), a kappa-receptor agonist, induced an increase in pain threshold, when OFQ (0.1 or 1 microgram) was added together with one of them (except for the ith injection of DSLET), the increase of pain threshold was reduced obviously. CONCLUSION: OFQ induces hyperalgesia and antagonizes opioid analgesia mediated by mu- and delta-receptors in the brain and by mu- and kappa- but not delta-receptors in the spinal cord of rats.

Molecular study on the chromosome 15 breakpoints in the translocation t(15; 17) in acute promyelocytic leukemia (APL).

Chromosomal translocation t(15; 17) is a specific marker of acute promyelocytic leukemia (APL). In this study, molecular cloning of the t(15; 17) breakpoint was carried out in a Chinese APL patient. It has been shown that the retinoic acid receptor alpha (RARA) gene, normally located on chromosome 17, was fused with a new transcription unit PML, normally localized on chromosome 15. We have subsequently cloned a portion of the PML gene and generated a panel of probes. A PML gene rearrangement was detected in 33 out of 36 APL cases studied. 24 rearrangements were clustered in a 4.4 kb region, designated here as PMLbcr1 whereas 9 rearrangements were concentrated in a 6.5 kb region, defining another breakpoint cluster region (PMLbcr2). These two types of rearrangement constitute the basis for the heterogeneity of the PML-RARA fusion gene and its possible biological significance remains to be explored.

Inhibitory property characterization and reactive site exploration of the arrowhead proteinase inhibitor.

Affinity chromatography was used to separate two components A and B of the crystalline arrowhead proteinase inhibitor. Both A and B are double-headed and multifunctional proteinase inhibitors. Inhibitor A is capable of inhibiting equimolarly trypsin and chymotrypsin simultaneously, and has a weak inhibitory activity toward kallikrein; whereas inhibitor B can inhibit two molecules of trypsin simultaneously, and shows rather higher inhibitory activity toward kallikrein than inhibitor A, but its inhibitory activity toward chymotrypsin is much weaker than that of inhibitor A. The results of chemical modification and the competitive binding of trypsin and chymotrypsin with inhibitor A showed that the two reactive sites of both inhibitors A and B are Lys and Arg residues. Among them the Lys reactive site is specific for inhibiting mainly trypsin, whereas the active domain composed of the Arg reactive site appears to be multifunctional and capable of inhibiting many different Ser proteinases. Based on the structural characteristics of inhibitors A and B, it was predicated that the two reactive sites should be located in the positions Lys-Ser (44-45) and Arg-Tyr-Lys (76-78), respectively. In inhibitor A, there exists another hydrophobic residue involved in inhibiting chymotrypsin. This residue might be situated in the reactive region composed of the Arg reactive site.

Total synthesis of Trichosanthes trypsin inhibitor and its analogue.

Trichosanthes trypsin inhibitor (TTI) is a peptide consisting of 27 amino acid residues with three pairs of disulfide bonds. This paper reports the total synthesis and disulfide bond refolding of this inhibitor and its analogue. After purification, the amino acid sequence and stoichiometrical inhibitory activity against trypsin of the synthetic inhibitor were compatible with those of the natural inhibitor. The analogue of this inhibitor in which residue Met in position 6 was replaced by Ala was also synthesized. The antitrypsin activity of this synthetic analogue was also approximate to that of the natural inhibitor.

Studies on natural and modified peptide Trichosanthes trypsin inhibitors.

A peptide trypsin inhibitor was isolated and purified from the roots of Trichosanthes kirilowii (a Chinese medical herb) by using immobilized anhydro-trypsin affinity chromatography and HPLC C18 column reverse chromatography. It contains two major components, both consisting of 27 amino acid residues with three pairs of disulfide bonds. The sequence determination indicated that the difference between them is only in the ninth position, being Gln and Lys, respectively. The peptide bond of the inhibitor reactive site Arg-Ile (3-4) is easy to cleave at low pH by trypsin, resulting in a modified inhibitor. It might be the smallest naturally occurring protein inhibitor so far known. The modification reaction of the Trichosanthes inhibitor with trypsin is similar to the catalytic enzyme-substrate reaction. The dissociation constant of the modified inhibitor with trypsin is around fourfold that of the natural inhibitor.

Crystal structure determination of mung bean trypsin inhibitor Lys fragment-bovine trypsin complex--molecular replacement, electron density map at 3.0 angstron resolution.

The orientation and position of the trypsin molecule in the complex crystal cell mung bean trypsin inhibitor Lys fragment (MBILF)-bovine trypsin (BTRY) have been successfully determined by molecular replacement method with the model of the refined bovine trypsin molecule. Starting from the BTRY coordinates which were oriented and located in the correct azimuth and position in the complex cell according to the result from rotation function and translation function, sim-weighted Fourier map with coefficients 2/Fo/-/Fc/ at 3.0 A resolution was calculated. Besides the electron density which is obviously attributed to itself, in the vicinity of the active site of BTRY the dense contour levels corresponding to the MBILF and and its boundary could be clearly seen in this map. The size of MBILF was approximately estimated at 15 x 15 x 25 A.

The crystallographic study of the complex of the Lys active fragment of the mung bean trypsin inhibitor with bovine trypsin.

The Lys fragment of mung bean trypsin inhibitor can combine with bovine trypsin to form a complex at an equal molar ratio. The single crystals of the complex were obtained by using the micro-still-setting method and the X-ray diffraction extended to 1.8A resolution. Its space group is P212121 with cell dimensions a = 62.9(1)A, b = 63.4(1)A and c = 69.7 (2)A. There is one complex molecule in a crystallographic asymmetric unit.

Studies on the mung bean trypsin inhibitor--reduction and reoxidation of the disulfide bonds of the Lys active fragment.

Two peptide chains A1 and A2 of the Lys active fragment, linked via a couple of inter-disulfide bonds, could be separated from each other after reduction with dithiothreitol and gel filtration on Sephadex G-25. Reoxidation of the reduced peptide chain A1 resulted in recovering the inhibitory activity with 25% yield, based on the original activity of the Lys fragment. The A1 active fragment was further purified by affinity chromatography with immobilized trypsin. Sephadex G-25 gel filtration produced two forms of the A1 active fragment, the major fraction being a monomer and the minor one being a dimer with lower activity. The results obtained offered evidence of the evolution of mung bean inhibitor from an ancestral single-headed inhibitor by fused gene duplication with A2 as a connecting peptide. The CD spectra of the Lys fragment and the reoxidized peptide chain A1 were also compared.

Studies on bradykinin potentiating peptide (BPP) from venom of Zhejiang pit viper (Agkistrodon halys pallas)--relationship between structure and function of BPP.

The structure of the bradykinin potentiating peptide component BPP1 from a 70% methanol extract of the lyophilized powder of pit viper from Zhejiang Province is shown as follows: (formula see text) When the pyroglutamyl residue was removed by pyroglutamate aminopeptidase, the activity of BPP1 was enhanced two-fold but decreased gradually during the course of Edman degradation. The above facts show that the structure integrity of the C-terminal is absolutely necessary for the activity of BPP1.