Objective To establish a method of GS-MS for determination of polychlorinated biphenyls (PCBs) in human cerumen. Methods The PCBs in cerumen samples was separated by n-hexane extraction, liquid-liquid extraction separation and column chromatography purification, then was analyzed quantitatively by gas chromatography and affirmed qualitatively by mass spectroscopy. Results The detection limit, average recovery rate and precision of this method were 0.15 ?g/g fat, 92.1% and 3.0%-4.6% (n=12) respectively. The contents of PCBs in 90 cerumen samples collected in polluted and control areas were detected. The levels(0.00-18.42 ?g/g fat) of PCBs in cerumen collected in polluted area were significantly higher than that in the control area(P

Objective To investigate the distribution and environmental pollution of dioxin-like polychlorinated biphenyls(PCBs) in a place of disused transformer and other electronic waste disassembled. Methods The ambient air, aerosol, water, soil and vegetable samples were collected to analyze the dioxin-like PCBs level with isotope internal standards and HRGC-HRMS techniques. Results The concentration of dioxin-like PCBs in ambient air was 4 031.77-4 942.34 pg/m3(namely 1.832-2.666 pg TEQ/m3), while in aerosol it was 325.45-3 499.77 pg/m3(0.370 9-1.536 3 pg TEQ/m3). In the soil sample, the value was 2 316.59-19 100.30 pg/g(1.358-15.264 pg TEQ/g) dry weight. In water it was 5.731-237.640 ng/L(2.585-73.046 pg TEQ/L). In vegetable it was 1 078.45-1 620.04 pg/g(0.778 3-1.126 2 pg TEQ/g) wet weight. In all air, aerosol, soil, water and vegetable samples, the content of PCB-118 was the most. PCB-126 had the highest TEQ value. In non-ortho PCBs, PCB-77 was of maximum content. In mono-ortho PCBs, PCB-118 was the most, and the followings were PCB-105, PCB-156 and PCB-167. All the 12 kinds of PCBs were investigated in all samples except PCB-169 absent in ambient air. Conclusions The investigated area around the disassembly of obsolete solid waste has been badly contaminated by dioxin-like polychlorinated biphenyls(PCBs), and PCBs already exist in the vegetables. According to the results of the present paper, some further studies should be carried out to evaluate the toxic effects on food and people health.

Objective To construct a fusion minigene expression vector of seven HCV NS3 epitopes and express the fusion gene in the eukaryotic cell line, in order to form a foundation for the investigation of the DNA immunization for HCV prevention. Methods Three pairs of complementary oligonucleotides primers covering all seven HCV NS3 epitopes were synthesized. Overlapping extension PCR were performed to construct the fusion minigene and cloned in pEGFP-N3 and pBuDCE vector respectively. The fusion protein was detected by Western blotting and flow cytometric analysis. The transfection efficiency was assessed with by flow cytometric analysis. Results The HCV NS3 epitope fusion minigene was obtained and inserted into pEGFP-N3 and pBuDCE vector respectively. The recombinant plasmid pEGFP-DR4 and pBuDCE-DR4 were expressed in 293T and 046W cell lines respectively. The Western blot result indicated that the expected 36 kDa minigene/EGFP fusion product was expressed from pEGFP-DR4, while 13kDa product from pBuDCE-DR4 respectively. Conclusion The HCV NS3 epitope fusion minigene was expressed in the eukaryocyte expression system.

Objective To verify the validity and feasibility of national early warning score (NEWS) in evaluation of death risk in elderly patients with critical illness,in order to find out which scoring method is more suitable for elderly critical illness patients.Methods A prospective case-control study was conducted.The critical illness patients aged over 60 years old with the length of hospital stay over 24 hours,and admitted to Department of Emergency of Qingdao Municipal Hospital from January to December 2015 were enrolled.The clinical data including in emergency and the actual outcome of patients were collected,and the patients were divided into death group and survival group according to 30-day outcome.Patients in the two groups were assessed by using NEWS and risk classification according to the first results of vital signs monitoring.Multivariate logistic regression model was used to analyze the relationship between the NEWS classification and the risk of death in elderly critical ill patients.Results 1 950 emergency elderly patients with critical illness were enrolled,with 78 cases (4.0％) dead within 30 days and 1 872 survived (96.0％).Compared with the survival group,patients in death group were older (years:79.8 ± 10.8 vs.75.3 ± 8.9,t =4.335,P ＜0.001),and had higher acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (22.9± 4.6 vs.18.2 ± 4.8,t =8.487,P ＜ 0.001),lower Glasgow coma scale (GCS) score (12.2 ± 4.5 vs.13.4 ± 5.2,t =-2.007,P =0.045),higher incidence of respiratory system diseases (29.5％ vs.17.9％,x 2 =12.742,P =0.013),higher NEWS score (11.2 ± 5.5 vs.3.9 ± 2.7,t =22.063,P ＜ 0.001),as well as higher proportion of patients with NEWS classification of high risk and very high risk (65.4％ vs.15.8％,x 2 =263.125,P ＜ 0.001).With the increase of NEWS risk classification,mortality rate was also increased,and the mortality rate in the patients with low,medium,high and very high risk were 0.81％ (9/1 108),3.63％ (18/496),5.83％ (13/223),30.89％ (38/123),respectively,with statistically significant difference (x 2 =179.741,P ＜ 0.001).It was showed by logistic regression analysis that the NEWS score of elderly patients with critical illness were positively correlated with 30-day death.The 30-day death risk of patients with middle risk,high risk and very high risk was 4.600,9.052 and 54.598 folds of the patients with low risk respectively.Conclusion NEWS score can be used to assess the risk of death in emergency elderly patients with critical illness.NEWS risk classification can quantify and classify the risk of death in the elderly patients with critical illness.

Objective To investigate the clinical and pathological characteristics of light chain proximal tubulopathy (LCPT).Methods Nine patients with LCPT diagnosed by renal biopsy in Peking University First Hospital from January 1,2011 to September 30,2016 were enrolled,and their clinical findings and pathological features were reviewed.Immunofluorescence (IF) of light chains (κ,λ) on paraffin sections after protease digestion and immunogold labeling of light chains (κ,λ) on ultrathin sections were performed in some cases.Results The main clinical manifestation of the nine patients was proteinuria of small molecules,with acute or chronic renal insufficiency,and six of them led to partial or complete Fanconi syndrome (FS).The hematologic diseases included 3 cases of multiple myeloma and 6 cases of monoclonal gammopathy of renal significance (MGRS).Pathological examination of renal biopsy showed two types:crystalline and noncrystalline LCPT.Seven cases of crystalline LCPT were stained for κ light chain,the proximal tubular epithelial cytoplasm exhibited fine granular vacuolation,with needle-shaped crystals and clear clefts by light microscopy,the intracytoplasmic inclusions of various shapes including rhomboidal,rectangular and rod-shaped crystals were identified by electron microscopy.Two cases of noncrystalline LCPT were stained for λ light chain,the prominent argyrophilic granules in cytoplasm of proximal tubular epithelia were observed by light microscopy,and intracytoplasmic large and irregular shaped phagolysosomes were found by electron microscopy,cast nephropathy were coexisted in these 2 cases,the additional light chain deposition disease were confirmed in one of them by electron microscopy and IF.All cases had monotypic staining of light chains in cytoplasm of proximal tubules by IF on frozen tissue and paraffin sections after protease digestion,with the latter method being more sensitive than the routine IF.The immunogold labeling showed specific monotypic labeling of κ and λ light chain on intracytoplasmic crystals and phagolysosomes respectively by immunoelectron microscopy.Conclusions LCPT is a rarely reported entity that manifested as acquired Fanconi syndrome and dysfunction of proximal tubules clinically.Pathologically it is divided into two types:crystalline and noncrystalline LCPT,with more prevalent of κ light chain related crystalline type,noncrystalline LCPT is mostly λ type,and is easily coexisted with cast nephropathy.The IF and immunoelectron microscopy of light chains(κ,λ) and ultrastructural examination by electron microscopy are important methods for the diagnosis of LCPT.

Purpose To investigate the clinicopathological features of adolescent mixed epithelial and stromal tumor of the kidney ( MESTK) and improve recognization of this rare disease. Methods Clinicopathological and immunohistochemical characteristics of MESTK occured in 17-year-old girl were studied. Reviewed the related literatures, clinical and pathological characteristics of adoles-cent MESTK were analysed comprehensively. Results Congenital perineal spill was the main clinical manifestations. Microscopically, the tumor showed nodules and was composed of a mixture of epithelial and stromal elements. Glands were lined with columnar or cilia-ted columnar cells. Stromal cells surrounding glands seemed like ovaries and away from glands seemed like the differentiation of smooth muscle. Immunohistochemical staining revealed that the epithelial cells were positive for CK7 and vimentin. Stromal cells expressed desmin, smooth muscle actin ( SMA) , ER and PR. It was noteworthy that stromal cells away from glands expressed desmin. Conclu-sions As a kind of rare benign neoplasm of kidney, MESTK often occurs in perimenopausal women. But MESTK can also occur in ad-olescence, and has nothing to do with the use of hormone. Therefore, the renal tumor occurred in teenagers with biphasic differentiation should be differentiated from MESTK.

Objective To evaluate the efficacy of modified quadruple therapy for patients who were failed in previous Helicobacter pylori ( Hp) eradication treatment .Methods A total of 86 patients with confirmed Hp infection and failed in previous Hp eradication treatment were collected from Qingdao Municipal Hospital during January 2012 and January 2014.Patients were randomly assigned into two group:43 patients in control group were given conventional quadruple therapy ( rabeprazole +colloidal bismuth pectin +amoxicillin +clarithromycin for 14 d ) , and 43 patients in test group were given modified quadruple therapy ( rabeprazole +colloidal bismuth pectin +amoxicillin +clarithromycin for 7 d, and lansoprazole +colloidal bismuth pectin +levofloxacin +metronidazole for 7 d).Chi square test was performed to analyze per-protocol (PP) eradication rates, intent-to-treat (ITT) eradication rates, and Hp recurrence rates between two groups .Results Among 43 patients in test group , 42 completed treatments with PP eradication rate of 100.00% and ITT eradication rate of 97.67%.All patients in control group completed treatments , and Hp eradication was observed in 24 patients , and both PP and ITT eradication rates were 55.81%.The differences in PP and ITT eradication rates between two groups were of statistical significance (χ2 =23.90 and 21.11, P<0.05).The 3-month and 6-month Hp recurrence rates in test group (14.29%and 20.00%) tended to be lower than those in control group (20.83%and 34.78%), but the differences were not of statistical significance (χ2 =0.12 and 1.68, P>0.05).Conclusion The efficacy of modified quadruple therapy for patients who were failed in the previous Hp eradication treatment is satisfactory.

Objective To elucidate the clinicopathological and hereditary characteristics in Fabry nephropathy.Methods The clinical and pathological features of 14 patients with Fabry nephropathy were collected.The activities of α-Gal A were measured in 4 probands.Screenings of GLA mutations were done in 12 patients.Results The ratio of Fabry nephropathy in the patients with renal biopsy was 0.074 ％ (14/19 005),the average diagnostic age was (30.57±9.32) years,male to female ratio was 2.5∶ 1.All the patients had proteinuria,and the median of urine total protein (UTP)was 1.71 g/24 h (0.32-4.71 g/24 h).Two of them got nephrotic proteinuria,5 of them got microscopic hematuria,4 of them got renal function insufficiency.Angiokeratomas was the most common manifestation (10/14),followed by cardiac involvement (6/14).Hypohidrosis and diseases of central neural system could also be seen in these patients.There were 9 classic phenotype,the remaining 5 were variants/renal variants.The family information was collected in 10 pedigrees,6 of them had family histories of kidney disease.Renal pathology showed vacuolization of glomerular visceral epithelial cells was the prominent feature,global and segmental glomerulosclerosis were seen in some patients by light microscopy.The myelin bodies or zebra bodies were identified in podocytes by electron microscopy,but only were found in some podocytes of 2 females.The activity of α-Gal A was decreased compared with the normal range in 4 probands.The mutations of GLA were demonstrated in 11 patients.Three novel mutations of GLA gene were identified,which were all deletion/insertion mutations with classic phenotypes.Most variants carried the mutations located in the buried/partial buried areas of α-Gal A (3/11).The classical phenotype carried GLA mutations as W162X,F169S,S201F,N272K,L310R,while variant phenotype carried GLA mutations as I91T,R112H,Q312H.Conclusions The ratio of Fabry nephropathy in patients with renal biopsy is 0.074％.Angiokeratomas and cardiac involvement are often accompanied with Fabry nephropathy.The genotypes of GLA may have close relationships with the phenotypes of Fabry nephopathy.

Objective:To investigate the clinicopathological characteristics of renal light and heavy chain amyloidosis (AHL).Methods:Ten patients with renal AHL diagnosed by renal biopsy in Peking University First Hospital and Institute of Nephrology of Peking University from January 2015 to June 2020 were enrolled. Clinicopathological data of these patients was collected and reviewed.Results:AHL typically affected older patients, with a male/female ratio of 7:3. The clinical manifestations were mainly edema and heavy proteinuria. At the same time, 7/10 of patients presented with nephrotic syndrome, 7/10 presented with microscopic hematuria, and 3/10 presented with renal insufficiency. Laboratory examinations showed monoclonal immunoglobulin in blood and urine in all patients, and IgGλ was the most common one (5/10). Decreased serum complement could be seen in some patients. The ratio of serum free κ light chain and free λ light chain was abnormal in all patients who underwent serum free light chain test. None of the 10 patients met the diagnostic criteria of multiple myeloma. Except for one of the 10 patients who was diagnosed as Waldenstrom's macroglobulinemia, the rest were diagnosed as monoclonal gammopathy of renal significance (MGRS). Bone marrow of 2/6 of patients were positive for amyloid. Cardiac involvement was confirmed in only one patient. Renal biopsy demonstrated amorphous eosinophilic material, which was Congo red positive, was deposited in glomerular mesangial area (10/10), capillary vessels (8/10), renal interstitium (9/10), peritubular capillary walls (9/10) and arterioles (8/10). This material showed apple green birefringence under polarized light. Immunofluorescence showed that single heavy chain and single light chain were positive at the same time, which was consistent with the results of mass spectrometry analysis. Ultrastructural evaluation revealed randomly oriented, non-branching fibrils with a diameter of 8-12 nm.Conclusions:Main clinical manifestations of AHL amyloidosis are edema and massive proteinuria, along with a high incidence of hematuria, a low portion of heart involvement and high frequency of whole molecule of monoclonal immunoglobulin (IgGλ dominant) by serum immunofixation electrophoresis. Renal pathology shows the commonly involved kidney compartments of amyloid deposits are glomerular capillary walls and peritubular capillary walls in patients with AHL amyloidosis.

Objective To study interleukin-23 (IL-23) levels in serum and dendritic cells of acute-on-chronic liver failure (ACLF) patients with chronic hepatitis B (CHB) and to explore its relationship with the prognosis.Methods Peripheral blood mononuclear cells and serum were collected from 40 ACLF patients with CHB (including survival group 27 cases and non-survival group 13 cases) and 26 healthy controls.Monocytes were induced to immature dendritic cell in vitro and TNF-α was added to induce dendritic cell maturation.IL-23 mRNA of dendritic cells was detected by real time polymerase chain reaction (PCR), and serum IL-23 level was measured by enzyme-linked immuno sorbent assay (ELISA).Differences among the parameters with normal distribution were compared using t test, those with non-normal distribution were compared using non-parametric Mann-Whitney U-test, and the relationship between two variables was assessed by Spearman′s rank correlation.Results International normalized rate (INR) and model for end-stage liver disense (MELD) scores in non-survival group of ACLF were higher than those in survival group (INR: 2.32 vs 1.64, U=69.00, P=0.002 2;MELD:36 vs 30, U=64.50, P=0.001 4).However, there were no significant differences between two groups at gender, age, alanin aminotransferase (ALT), aspartate aminotrans ferase (AST), bilirubin, creatinine, hepatitis B virus (HBV) DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and serum IL-23.IL-23 mRNA level in dendritic cells at baseline in non-survival group of ACLF was significantly higher than that in survival group (76 vs 43, U=71.50, P=0.002 8).After treatment, serum IL-23 was significantly declined in survival group ([160±75] ng/L vs [91±49] ng/L, t=4.012, P=0.000 2), but not in non-survival group.Significant positive correlation was observed between IL-23 mRNA level in dendritic cells and MELD score at baseline (r=0.7198,P<0.01).Conclusions Persistent high serum IL-23 level suggests poor prognosis in ACLF patients with CHB.IL-23 mRNA expression in dendritic cells has good consistency with MELD score and the patients with high IL-23 mRNA expression has poor outcome.