A method was developed for purification of 20S proteasome (20S core particle, CP) by combining differential centrifugations with nondenaturing polyacrylamide gel electrophoresis (native-PAGE), irrespective of species origins of CPs. CP purified from human erythrocytes was subjected to proteomic analysis by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), revealing 33 spots of subunit isoforms with different molecular weights and isoelectric points, more than 14 constituent subunits. Furthermore, other four CPs were purified from yeast, mouse liver, two pancreatic cancer cell lines SW1990 and PANC-1 using this method mentioned above, and subjected to proteasome heterogeneity analysis by native/SDS-PAGE (native/sodium dodecyl sulphate polyacrylamide gel electrophoresis), together with CP from erythrocytes. The method described acts as a rapid and effective tool for CP isolations, and the results obtained may be served as a footstone for the investigations of species-dependent proteasome heterogeneity.

Objective To obtain a reliable estimate of the risk of H. pylori infection in causing pancreatic cancer ,by perform‐ing a M eta‐analysis of the existing observational studies evaluating the association .Methods Observational studies comparing the prevalence of H. pylori infection in patients with pancreatic cancer and healthy controls were identified through systematic search in the Medline ,EMBASE ,the Cochrane ,PubMed ,VIP database .H. pylori infection was confirmed by serological testing using an anti‐gen‐specific enzyme‐linked immunosorbent assay .Pooled adjusted odds ratios (AOR) and associated 95% confidence intervals (CI) were obtained by using a Dersimonian and Laird random‐effects model .Results Six studies involving a total of 2 335 patients met our eligibility criteria .A significant association between H. pylori seropositivity and development of pancreatic cancer (AOR=1 .38 , 95% CI:1 .08-1 .75 ;P=0 .009) was seen .No significant association had been seen on pooled analysis of the three studies assessing the relationship between CagA positivity and pancreatic cancer (AOR=1 .14 ,95% CI:0 .66-1 .97 ,P=0 .639) .Conclusion The da‐ta suggests an association between infection with H. pylori and the development of pancreatic cancer .Further research is needed to confirm our findings .

Objective There is a lack of information on the effect of hypoxia on the virulence of Cryptococcus neoformans . This study was to construct a Cryptococcus neoformans bilfilm model in vitro and observe the influence of hypoxia on the biofilm forma-tion. Methods We constructed a Cryptococcus neoformans biofilm model in vitro in 24-well and 96-well microculture plates using DMEM culture medium at the oxygen concentrations of 21%( normoxia ) and 1% ( hypoxia ) .We collected the Cryptococcus neofor-mans biofilms at 2, 4, 8, 12, 24, 48, and 72 hours after culturing and observed their thickness , structure, and growth activity in the two different oxygen conditions by light microscopy , confocal laser scanning microscopy , and MTT assay . Results The Cryptococcus neoformans biofilm model was successfully constructed in the conditions of both hypoxia and normoxia .The processes of biofilm forma-tion in the two conditions were similar , involving adhesion , aggregation , micro-colony formation , and biofilm maturation , with the ulti-mate biofilm thickness of about 16 μm.The cell density and growth activity of the biofilms increased with the extension of incubation time, gradually stabilized with their maturity , and both were relatively higher at 1%than at 21%oxygen concentration . Conclusion The abilities of Cryptococcus neoformans biofilm formation vary with different oxygen concentrations , and hypoxia can promote the for-mation of Cryptococcus neoformans biofilms .

Recurrent laryngeal nerve injury is one of the serious complications after thyroidectomy.Unilateral injury causes hoarseness,while bilateral injury causes difficulty in breathing or even life-threatening glottis obstruction.Analyzing the root cause of the thyroid injury,firstly it is the anatomical factors of recurrent laryngeal nerve,namely the close and complex relationship between the recurrent laryngeal nerve and the inferior thyroid artery,the existence of branches of recurrent laryngeal nerve and its variation,and the presence of non recurrent laryngeal nerve and Zuckerkandl nodules.Those are all made the recurrent laryngeal nerve easy to be damaged.Secondly it is because of the vulnerability of the recurrent laryngeal nerve itself.Last improper using of energy operation instrument will cause heat injury on nerves.Below counter measures can be implemented to prevent recurrent laryngeal nerve injury.Dissect to show recurrent laryngeal nerve or make it ‘ visualization’ during thyroidectomy.Elaborately anatomize recurrent laryngeal nerve to appropriate degree.Be familiar with the property of energy operation instrument and thus safely use them to reduce the heat injury of recurrent laryngeal nerve.Reasonably use the intraoperative nerve monitoring in the surgery,which assist to reduce the risk of injury of recurrent laryngeal nerve.

Objective To investigate the heat effects of the high-frequency electric knife on the recurrent laryngeal nerve ( RLN) in pigs and the safety margin in which electric knife can be used .Methods Totally 12 pigs for experiment were randomly divided into 3 groups by the distance between the head of the electric knife and the nerve(2 mm, 1 mm, 0 mm).The application time of the electric knife touching RLN was set to be 3 s and the application energy of the electric knife was 90 W.The data of electromyogram were measured by means of nerve detector before and after operation .Statistical analysis was made based on those data .The nerve tissues were taken to make paraffin sections so that the histological change can be observed and compared before and after damage.Results The data from electromyogram by nerve detector indicated that the difference of the amplitude of group 2(1 mm)and group 3(0 mm)had statistical significance(P<0.05)within group.There were significant difference of the amplitude between the 3 groups.Histological study showed that the tissues of group 3(0 mm)had obvious injury .Conclusions In thyroid surgery , the safety range of high frequency electrical knife used around RLN is:the distance from electrical knife head to nerve should be no less than 2 mm.

Objective To analyze the risk factors of incision infection and spinal canal hematoma after lumbar spinal posterior surgery.Methods Data of 33 patients with incision infection and 25 patients with spinal canal hematoma after posterior surgery for lumbar spinal disease from January 2010 to December 2014 were retrospectively analyzed.For the patients with incision infection,20 of them were males and 13 were females,with an average age of 58.85±8.76 years (range 38-79 years).Of these patients,5 were diagnosed with lumbar disc herniation,9 with lumbar spondylolisthesis,15 with lumbar spinal stenosis and 4 with lumbar vertebral fracture.For patients with spinal canal hematoma,17 of them were males and 8 were females,with a mean age of 60.96±11.37 (range,38-77).The diagnoses of them were lumbar disc herniation in 18 patients,spondylolisthesis in 3 patients and spinal stenosis in 4 patients.From the same period database,patients who underwent lumbar posterior lumbar surgery with no postoperative complications were selected by 3:1 ratio as the control group according to age,gender and diagnosis.Results For patients with incision infection,it was found that diabetes mellitus,preoperative RBC,hemoglobin,total protein,albumin,serum calcium,operation time,number of segment fusion,intraoperative blood loss,postoperative WBC,RBC,hemoglobin and platelet were significantly different from those with non-infection group.Moreover,multivariate logistic analysis showed that diabetes mellitus (OR=3.716,P=0.032),preoperative serum calcium (OR＜ 0.001,P=0.001),intraoperative blood loss (OR=1.002,P=0.014),postoperative hemoglobin (OR=0.923,P=0.018) and postoperative platelet (OR=1.007,P=0.017) were independent risk factors for postoperative incision infection.For patients with spinal canal hematoma,it was found that patients' preoperative total protein,albumin,serum calcium,platelet,operation time,intraoperative blood loss and postoperative total protein were significantly different from non-hematoma group.Multivariate logistic analysis showed that preoperative serum calcium (OR＜ 0.001,P=0.001),preoperative total protein (OR=1.298,P=0.043),intraoperative blood loss (OR=1.003,P=0.021) and postoperative total protein (OR=1.080,P=0.028) were independent risk factors for postoperative spinal canal hematoma.Conclusion The preoperative diabetes mellitus,serum calcium,intraoperative blood loss,postoperative hemoglobin and platelet were important risk factors for lumbar incision infection.And preoperative serum calcium,total protein,intraoperative blood loss and postoperative total protein were the risk factors for spinal canal hematoma.

Objective To study the expression of β-catenin and cyclin-D1 in sporadic parathyroid adenomas and the clinical significance.Methods Immunohistochemistry and RT-PCR were used to examine the expression of β-catenin and cyclin-Dl in 20 cases of sporadic parathyroid adenomas,10 cases of parathyroid hyperplasia tissues and 8 cases of normal parathyroid tissues respectively.Results The results of immunohistochemistry showed that the expression of β-catenin was decreased on cell membrane of normal parathyroid tissues,parathyroid hyperplasia tissues and sporadic parathyroid adenomas,and was increased in the cytoplasm and the nucleus.The expression of cyclin-Dl was significantly higher in the adenoma group and hyperplasia group than in the normal group( P ＜ 0.05 ).The difference had no statistical significance in terms of cyclin-Dl expression between the adenoma group and the hyperplasia group( P ＞0.05 ).The abnormal expression of β-catenin was significantly correlated with the overexpression of cyclin-D1 in sporadic parathyroid adenomas( P ＜ 0.05 ).RT-PCR analysis showed that the expression of cyclin-D1 mRNA was 2.36 ± 1.12 vs 1.50 ± 1.03 ( P ＜ 0.05 ),and the expression of β-catenin mRNA was 1.02 ± 0.45 vs 0.88 ± 0.56( P ＞ 0.05 ) in adenomas and normal parathyroid tissues respectively.Conclusion The abnormal expression of β-catenin activates cyclin-Dl and thus leads to the uncontrolled cell proliferation and differentiation,which may be one of the mechanisms of the occurrence of sporadic parathyroid tumors.

Objective There have been a few reports on chronically recurrent and generalized superficial mycosis caused by trichophyton rubrum.This article was to investigate the cause, diagnosis and therapy of the mycosis. Methods 5 patients with chron-ically recurrent and generalized superficial mycosis caused by trichophyton rubrum were collected from June 2012 to June 2014 in our hospital.Bacterioscopic examination and cultivation were made on skin lesions of the patients.A typical patient who had 7-year course of desease with toenails seriously infected and widespread skin eruption was selected for histopathology examination on skin lesions, mi-crobiology and molecular biology study on 4 bacterial strains isolated from skin lesions in different parts, and in vitro chemosensitivity assay for drug selection.PCR (rDNA ITS sequence analysis) was performed for diagnosis and early treatment. Results Microscopic examintion on skin lesions demonstrated numerous septate, branched hyphae.Cultivation and molecular biology study identified tricho-phyton rubrum.The strain was identified as trichophyton rubrum by ITS sequence analysis and the isolated strains from different lesions were the same fungal species.Histopathology examination revealed slight hyperplasia of squamous epithelium , epidermal hyperkeratini-zation and the upper dermis presented a sparse perivascular lymphocytic infiltrate.The PAS-stain confirmed the presence of few hyphae in the horny layer.The pathogen of this case was trichophyton rubrum. A combination therapy with systemic itraconazole and topically applied terbinafine hydrochloride cream was successful.A follow-up examina-tion one year later showed no recurrence of symptoms. Conclusion The isolation and identification of pathogen is the key to the diagnosis of chronically recurrent and generalized superficial mycosis, with ad-ditional attention to all or none toenail infection.The therapy should not focus simply on the tinea corporis, while comprehensive treatment combined with chemosensitivity assay is preferred.

To detect furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone ( AMOZ ) in fish sample, an Eu3+ labeling time-resolved fluoroimmunoassay ( TRFIA ) was developed. The effects of experimental conditions including AMOZA-OVA concentration, dilution of antibody, and reaction time on the sensitivity of TRFIA were explored. The results showed that the optimized assay conditions were as follows:the AMOZA-OVA concentration was 0. 25 μg/mL; the antibody was diluted 5í104 folds, and the competitive reaction time was 50 min. Under optimal conditions, the method showed a detection limit of 0. 01 ng/mL, an IC50 of 0. 26 ng/mL and a linear range (IC20-IC80) of 0. 025-2. 83 ng/mL. The recoveries of AMOZ in fish at three spiked levels ranged from 78 . 0% to 86 . 0%, and the relative standard deviations were less than 15%. Good correlation between the ic-TRFIA and high performance liquid chromatography-tandem mass spectrometry was obtained for spiked food samples. The proposed ic-TRFIA method was suited for the determination of AMOZ residue in food samples.

Objective:To investigate the clinical efficacy in one stage reconstruction of composite defects of Achilles tendon and surrounding soft tissues with a flap transfer combined with allogeneic tendon transplantation.Methods:From July 2018 to August 2022, a total of 12 patients, including 9 males and 3 females, with a mean age of 31.5(ranged 8 to 56) years old, had surgery with flap transfer combined with transplantation of allogeneic tendon in one stage reconstruction for compound defects of Achilles tendon and soft tissue at the Department of Orthopaedics of First Affiliated Hospital of Nanchang University. The defects of Achilles tendons ranged from 4.0 to 9.0 cm, and the soft tissue defects sized from 3.0 cm × 4.0 cm to 14.0 cm × 6.0 cm. Of the 12 patients, 6 received transfers of sural neurovascular flaps, 3 with peroneal perforator flaps and 3 with free anterolateral thigh flaps(ALTF). The flaps sized from 4.0 cm × 4.5 cm to 15.0 cm×7.0 cm, and in addition, allogeneic tendon grafts were used to reconstruct the defects of Achilles tendons in all patients. All the flap donor sites were either directly sutured or covered with skin grafts. Follow-up was carried out by visits of outpatient clinic or telephone or WeChat distant interviews. The flap survival and recovery of ankle function and Achilles tendon were observed.Results:During the 3 months to 2 years of follow-up, none of the patient showed obvious immunological rejection against the transplanted allogeneic tendon. All 12 flaps survived well with the colour and texture close to the surrounding skin. No ulceration occurred in both of the donor and recipient sites. There was no re-rupture of the transplanted allogeneic tendon. At the final follow-up, ankle movement was measured at 13.4°±2.6° in dorsal extension and 33.6°±3.2° in plantar flexion. According to American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hind foot function score, a score of 88.7±5.6 was achieved with 7 patients in excellent, 4 in good and 1 was acceptable.Conclusion:In patients with a composite defect of Achilles tendon and surrrounding soft tissue, the application of a flap transfer combined with a homogeneous allograft tendon transplantation in an one stage surgery is a feasible surgical procedure. It can achieve a satisfactory outcome with less trauma and fewer complications.