Volume 93  ·   Issue 28  ·  2013

To explore the expressions of indoleamine 2, 3-dioxygenase (IDO) in hepatocellular carcinoma and analyze its relationship with clinicopathological parameters.

Quantitative real-time polymerase chain reaction (PCR), fluorogenic quantitative PCR, immunohistochemical and immunofluorescence were used to detect the expression of indoleamine 2,3-dioxygenase in hepatocellular carcinoma.

The IDO mRNA expression in cancerous tissues increased markedly than that in the corresponding non-cancerous tissues (2^-ΔΔCT=1.71, P=0.001).The immunohistochemical and immunofluorescence results showed that IDO protein was expressed in cytoplasm of hepatocellular carcinoma and tumor-surrounding tissues.But there was no expression in normal liver tissues from benign hepatic lesions and corresponding non-cancerous tissues.An overexpression of IDO protein was detected in 43 patients (48.3%), a low expression in 25 (28.1%) and no expression in 21 (23.6%).Relationship between IDO expression and clinicopathological parameters: an over-expression of IDO in HCC was associated with recurrence, survival time, metastasis and TNM stage (P<0.05), but not associated with patient′s cirrhosis, AFP level, histological differentiation type, Barcelona clinic liver cancer stage, gender, age, HbsAg positivity, number of tumors and tumor size (P>0.05).

An over-expression of IDO in HCC patients may affect patient prognosis.

To compare the clinical efficacies and safety of primary versus second liver resection for recurrent colorectal metastases to liver.

Between January 2000 and March 2011,a total of 126 patients underwent liver resections for metastases from colorectal cancer at our institution.Among these, 16 patients underwent repeat liver resections.The comparisons were made for primary and second liver resections in blood loss volume, complications, hospital stay, operative duration and 1,3, 5- year survival rates.

Compared with primary liver resection group, the 1, 3, 5-year survival rates of second liver resection group were 93.8%, 56.1% and 37.4% respectively.For second hepatectomy group, blood loss volume was (323.8±230.9) ml, operative duration (216.9±79.7) min, postoperative hospital stay (23.4±13.9) days and complication rate 18.8%.Compared with primary hepatectomy, there was no difference between two groups.

Second liver resection may provide long-term survival rates similar to those of primary liver resections.Repeat liver resection is warranted when potentially curative.

To explore the safety and feasibility of laparoscopic right hemihepatectomy via an anterior-inferior approach through retrohepatic tunnel in the dissection of short hepatic veins (SHVs).

After partial freeing of right liver, anterior peritoneum of inferior cava vena (ICV) was opened.Retrohepatic space was dissected via an anterior-inferior approach to establish the posterior tunnel partially.Then the first branch of right side SH Vs could be freed and ligated after its exposure through the right part of retrohepatic tunnel.The above procedure was repeated until the right side SHVs or the third hepatic portal became partially or completely blocked.If right side SH Vs were completely freed and ligament of right liver fully isolated, right hepatic vein could be exposed and ligated and selective blockage of the second hepatic portal blood flow accomplished.This technique was applied in 7 cases of laparoscopic right hemihepatectomy through curettage transaction and aspiration with laparoscopic Peng′s multifunctional operative dissector (LPMOD).

Six patients were treated successfully.In one case of right hepatic hemangioma, small margin auxiliary hematischesis was attempted because of troublesome hemostasis of middle hepatic vein branch.All of them underwent partial dissection of right side of SHVs.Two cases had complete dissection in which right hepatic vein was freed and ligated, the second hepatic porta blood flow controlled and right hemihepatectomy anatomically achieved.Operative duration was 300-540 min[mean, 399.1±74.7].The time of dissecting hepatic porta was 30-75 min[mean, 50.7±16.2].The time of dissecting SH Vs was 35-95 min[mean, 57.1±22.1].The time of liver transection was 60-160 min[mean, 115.9±32.3].Operative blood loss had a volume at 600-3000 ml[mean,1485.7±809.2].The postoperative hospital stay was 10-18 days[mean, 12.4±2.6].The postoperative time for ambulation, diet and flatus was 2-4, 1-4 and 2-4 days respectively.No severe postoperative complications occurred.

During laparoscopic right hemihepatectomy, dissecting SH Vs is both safe and feasible through a retrohepatic tunnel via an anterior-inferior approach.

To explore the electroencephalographic changes of patients with hyperphenylalaninemia (HPA) after dietetic treatments.

From July 2005 to February 2010, 34 HPA patients with abnormal electroencephalograms were recruited from China-Japan Friendship Hospital.Their electroencephalographic results and clinical features were compared before and after dietetic and antiepileptic treatments.There were 16 males and 18 females with an age range of 2 months to 8 years.All electroencephalographic results were abnormal.Among them, there were epileptic discharges (n=17), atypical sharp-slow wave complex (n=1) and slow wave background (n=16).All patients received a low intake of phenylalanine at 13- 50 mg·kg^-1·d^-1. Twenty patients had antiepileptic drug therapy.Before and after treatment for 3 months, 6 months, 1 year and 2 years, blood phenylalanine concentration, electroencephalogram, convulsion and other clinical parameters were observed.Before and after 1 year treatment, the tests of developmental quotient (DQ) or intelligence quotient (IQ) were performed.

The phenylalanine concentrations of the same patient group decreased significantly after 3 months, 6 months, 1 and 2 years after dietetic treatment ((0.51±0.39), (0.50±0.29), (0.59±0.42), (0.53±0.27) vs (1.33±0.64) mmol/L, all P=0.000).Among 19 epileptic cases, 16 ceased after treatment.And 14 stayed convulsion-free within 3 months post-treatments,2 cases had intermittent tonic-clonic seizures while there was 1 case of focal seizures.Fourteen cases withdrew antiepileptic medications or reduced their doses.After treatment, the electroencephalographic results became totally normal in 8 of the 19 epileptic cases.Electroencephalographic results improved in 5 cases versus pre-treatment.Spikes were controlled in 10 cases including 5 patients with hyperarrhythmia in the abnormal electroencephalographic group with epilepsy.Six cases continued abnormal electroencephalogram.In the abnormal electroencephalographic group without epilepsy (n=15),electroencephalograms were normal (n=3),improved (n=6) and continued mild abnormal waves (n=2).However,there were 4 cases with deteriorated electroencephalogram and 3 of them showed spikes.DQ (IQ) was evaluated in 16 cases.And variable mental retardation was observed in all of them.After,year treatment,7 cases were evaluated DQ (IQ), and their results showed improved (57±16 vs 50±16,t=-5.42, P=0.002).

Etiological treatment for HPA patients is important for controlling epilepsy and improving brain function.And electroencephalogram is useful for monitoring brain functions and evaluating treatment outcomes.

To explore the composition of bilateral urinary tract calculi so as to guide treatments.

A total of 126 patients with bilateral urinary calculi were recruited to undergo percutaneous nephrolithotomy or ureteroscopes lithotripsy.And 256 extracted stones were analyzed by infrared spectrophotometry.

The components of these stones were mainly mixture.The compositions included calcium oxalate (n=214,85%), carbonate apatite (n=113,45%), phosphate (n=43, 17%) and uric acid (n=28, 11%).And 63 patients had the same stone composition in bilateral urinary stones and 26 of them were of pure calcium oxalate, 2 phosphate while another 36 mixture.Different compositions were present in 60 patients with bilateral urinary stones.

The compositions of bilateral urinary stones are not always identical in a patient.The overall patient status should be considered if one side urinary stone is treated according to the composition analysis results of another side counterpart.

To discuss the role of direct lymphangiography in the diagnosis of primary chyluria.

Iodized oil was infused into lymphatic vessel of unilateral foot in 25 cases.The condition of lymphatic system imaging was dynamically observed via Innova 2100-IQ digital flatpanel angiography system.

All of them underwent direct lymphangiography.The results revealed unilateral (n=18) or bilateral (n=7) renal reflux of contrast media.Among 22 stenotic cases, there were partial (n=18) or whole (n=4) cervical section of thoracic duct, no imaging in cervicothoracic section of thoracic duct (n=1); thin imaging in cervical section and partial thoracic section (n=2); retroperitoneal lymphangictasia & tortuous lymphatic vessels (n=21); reflux of contrast medium to pelvic and/or contralateral sacroiliac part (n=13).

Capable of acquiring dynamic images of chyle (lymph) circulation, direct lymphangiography provides accurate estimation for the location, scope,extent of lesions of renal chylous reflux in primary chyluria so as to guide surgery.Stenotic cervical section of thoracic duct plays an important etiologic role in primary chyluria.

To explore the clinical characteristics and prognosis of patients with primary testicular non-Hodgkin′s lymphoma.

The clinical profiles and prognostic factors of 21 cases newly diagnosed as primary testicular non-Hodgkin′s lymphoma at Peking University Cancer Hospital from January 2005 to December 2012 were retrospectively analyzed.

Their median age was 59 (34-86) years.And they were classified as Ann Arbor stageⅠ (n=8), stageⅡ (n=2) and stageⅣ (n=11).There were B symptoms (n=4),extranodal involvement outside testis (n=12) and elevated lactate dehydrogenase (LDH) at diagnosis (n=6).The scores of international prognostic index (IPI) were 0-1 point (n=10), 3 points (n=10) and 4 points (n=1).The regimens included orchidectomy as the initial treatment (n=15), chemotherapy followed by radiotherapy (n=7) and CNS prophylaxis during treatment (n=15).All patients were pathologically diagnosed as diffuse large B-cell lymphoma.And 11 cases belonged to the non-germinal center B cell-like subgroup.First-line chemotherapy was either R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone) or CHOP-like-based regimen.Complete response was achieved in 85.7% of patients.The median follow-up period was 18 (6-58) months.The 1,2 and 3-year survival rates were 100%, 80% and 60% respectively.Statistical analysis showed that the first-line chemotherapy with rituximab was a prognostic factor (P=0.038).Other factors included stage (P=0.275), LDH level (P=0.179),β2-microglobulin level (P=0.229) and IPI (P=0.275).

The prognosis of primary testicular non-Hodgkin′s lymphoma is usually poor.The first-line chemotherapy with rituximab is a prognostic factor.

To evaluate the predicative value of serum parathyroid hormone (PTH) levels in outpatients of heart failure (HF) for hospitalization.

A total of 102 consecutive HF outpatients were enrolled.The receiver operating characteristic (ROC) curves demonstrated the optimal cut-off points of PTH levels for hospitalization due to HF.And Logistic regression analysis model was employed to analyze the independent association between PTH and hospitalization for HF.

The more advanced grade of New York Heart Association (NYHA), the higher serum level of PTH.The ROC curves showed PTH levels≥56.05 ng/L were the optimal cut-off point for hospitalization for HF with a sensitivity of 90.0%, a specificity of 89.2% and the area under ROC curve of 0.92.After adjustment for predictors for hospitalization due to HF (gender, age, diabetes mellitus, hypertension, left ventricular ejection fraction, estimated glomerular filtration rate and brain natriuretic peptide), PTH levels were associated with hospitalization due to HF (OR=1.282,95% CI 1.026-1.362).

The serum level of PTH in HF outpatients is an independent predicator for hospitalization due to HF.

To explore the clinical patterns and clinical significance for RET screening in adrenal pheochromocytoma (PHEO) associated with multiple endocrine neoplasia type 2A (MEN2A).

The clinical data of 32 PHEO patients with MEN2A from 13 unrelated MEN2A pedigrees from August 1989 to January 2013 were analyzed.The comprehensive medical data included systemic examinations and germline RET gene screening.

Among 68 patients belonging to 13 MEN2A families, 32 (47.1%) presented with PHEO.There were 19 males and 13 females with a mean age of (41±12) years.And the mean maximum diameter of PHEO was (4.6±2.2) cm.The diagnosis of PHEO was made after medullary thyroid carcinoma (n=12, 37.5%), simultaneously (n=12, 37.5%), initially (n=7,21.9%) and death during appendectomy for PHEO-induced hypertensive crisis (n=1, 3.1%).The diagnosis of PHEO was made before (n=22) or after (n=10) clinical screening.The former had 12 symptomatic cases while the latter only 1 case (12/22 vs 1/10,P=0.024).Except for 5 asymtomatic fatal cases during non-PHEO operations, bilateral PHEO was found in 17 cases including 3 unilaterally treated cases developing another PHEO in contralateral adrenal with a lag period of 5, 10 and 17 years.There were 7 symptomatic patients in bilateral cases versus 6 in unilateral cases (7/17 vs 6/10,P=0.440).Twenty-five patients underwent PHEO surgery: laparascopic approach in 14 cases (8 with bilateral simultaneous adrenalectomy) and open approach in 11 (2 with bilateral simultaneous adrenalectomy).And 10 patients undergoing bilateral adrenal-sparing operations or adrenalectomy required hormonal replacement therapy.During a mean observation period of 72 (1-282) months, no local recurrence, distant metastasis or Addisonian crisis were noted in 25 cases (contralateral relapse in 3 cases).Among them, 2 cases developed adrenocortical insufficiency unresponsive to an adjustment of hormonal doses.RET screening showed 4 recurrent missense substitutions in 32 MEN2A-PHEO patients: p.C634Y exon 11 (n=27, 84.4%), p.C634R exon 11 (n=3, 9.4%), p.C634F exon 11 (n=1, 3.1%) and p.C618R exon 10 (n=1, 3.1%).

The mutations of RET proto-oncognene of PHEO in MEN2A are frequently located at codon 634.A combination of pedigree examination and RET gene screening may facilitate an early diagnosis and early treatment of asymptomatic PHEO patients in MEN2A.Laparoscopic cortical-sparing adrenalectomy for preserving adrenocortical function is a preferred surgical approach.

To explore the polymorphism of cholesteryl ester transfer protein (CETP) gene -629C/A among the coronary heart disease (CHD) Han population of Tianjin area and evaluate the influences of genetic factors on atorvastatin therapeutic effects and clinical outcomes in pharmacogenomics and provide theoretical rationales for individualized treatment.

A total of 332 angiographically confirmed CHD patients at Tianjin Chest Hospital were recruited from October 2010 to July 2011.The CETP gene promoter polymorphism at position -629 was determined by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP).The serum level of CETP was determined by enzyme-linked immunosorbent assay (ELISA).Lipid levels were determined at baseline and 12 months post-treatment with 20 mg/d atorvastatin in all patients.Clinical follow-up were performed for more than 1 year (range, 12- 23 months).And major adverse cardiac events (MACE, including death, non-fatal infarction, revascularization and stroke) were analyzed.The Kaplan-Meier Log-rank test was used to compare MACE-free survival between different genotypes.

(1) The frequencies of variant -692A allele was 0.476, AA genotype showed reduced CETP levels and higher HDL-C levels compared with CC and CA genotypes.But it did not reach statistical significance (F=0.893, P=0.411 and F=1.279, P=0.282 respectively).Although a negative trend correlation existed between serum levels of HDL-C and CETP, it did not reach statistical significance (r=-0.151, P=0.081). (2) After 12-month therapy of atorvastatin, CCgenotype was shown to be associated with higher LDL-C, LP(a) reduction and HDL-C elevation in response to atorvastatin compared with CA and AA genotype. LDL-C levels decreased 43.5% in CC homozygotes, 25.5% in CA heterozygotes and 11.7% in AA homozygotes (P=0.001).HDL-C levels increased 9.2% in CC homozygotes,6.8% in CA heterozygotes and 5.5% in AA homozygotes.However the changes of HDL-C levels in three genotypes showed no significant difference (P=0.412). (3) There was a 7.83% incidence of MACE after a mean follow-up of (18.66±5.99) months.The outcomes were death (n=3, 0.90%), nonfatal infarction (n=7, 2.11%), revascularization (n=13, 3.92%) and stroke (n=3,0.90%).The cumulative MACE free survival rates were 96.2%, 92.1% and 87.3% in CC, CA and AA genotypes respectively (Log-rank P=0.444).

Variant AA genotype shows a higher level of HDL-Cs and a lowered level of CETP.However CC genotype offers a better benefit of statin therapy associated with lowered levels of LDL-Cand LP(a).And the long-term clinical prognosis is not affected among three genotypes.

To explore the relationship between the mRN A expression level of mitochondrial cytochrome oxidase and the maternal inheritance of asthma.

From January to December 2009, 220 asthma patients, 162 patient kins and 260 healthy subjects were recruited from Departments of Respiratory Critical Care Medicine and Pediatric Medicine at First Affiliated Hospital, Kunming Medical College.Lung function tests were performed and serum IgE level measured.The polymorphism of mitochondrial cytochrome oxidase gene polymorphisms was detected by direct sequencing.And the peripheral level of COX mRNA was measured by reverse transcriptionpolymerase chain reaction (RT-PCR).

No significant difference existed in age, gender among 3 groups.For 3 groups, the first second forced expiratory volume (FEV₁) /forced vital capacity (FVC) were 90.6±6.2, 92.3±2.3, 102.3±2.3 and FEV₁ percentage of expected value (FEV₁%) were (82.9±10.8)%, (94.8±5.4)% and (98.3±8.6)% respectively.The lung function was not significant difference among three groups.The m RNA expression level of mitochondrial cytochrome oxidase in peripheral blood were 0.357±0.217, 0.637±0.473 and 0.975±0.260 in the asthma, kin and control groups respectively.No significant difference existed in the expression level of COX3 mRNA among 3 groups (F=21.45, P=0.012).The serum level of lgE was the highest for the asthma patients.And it was significantly higher in the asthma group than that in the control group ((283.6±62.4) vs (52.3±13.7) μg/L, F=48.31, P<0.05).Moreover, the serum level of IgE was significantly higher in the kin group than that in the control group ((116.4±57.5) vs (52.3±13.7) μg/L, F=20.45, P<0.05).However, there was a negative correlation between the mRNA expression level of mitochondrial cytochrome oxidase and the serum level of IgE among 3 groups.

The down-regulated m RNA expressin of mitochondrial cytochrome oxidase may participate in allergic inflammation by regulating the level of IgE.And the maternal inheritance of asthma is in effect.

To evaluate the effects of down-regulated expression of transient receptor potential canonical (TRPC1) by RNA interference (RNAi) on proliferation and invasiveness of human lung adenocarcinoma cell A549 in vitro.

A549 cells were transfected with chemically synthesized small interfering RNA (siRNA) targeting TRPC1 gene.The mRNA and protein of TRPC1 were analyzed by realtime polymerase chain reaction (PCR) and Western blot respectively.To assess malignant phenotypes of transfected A549 cells, the assays of methyl thiazolyl tetrazolium (MTT), cell cycle and cell invasion were performed.

siRNA targeting TRPC1 dramatically suppressed TRPC1 expression.In vitro study showed that siRNA targeting TRPC1 significantly inhibited cell proliferation of A549 cells with an inhibitory rate of 34.7% while negative control siRNA had no effect on cell proliferation.Flow cytometric analysis showed that siRNA targeting TRPC1 increased the number of cells in G₀/G₁ phase (P<0.05).Moreover,a knockdown of TRPC1 expression effectively inhibited cell invasiveness in A549 cells (P<0.05).

Knocking down TRPC1 expression can inhibit proliferation and invasiveness of A549 cells in vitro.

To explore the therapeutic efficacy of double suicide gene system driven by carcinoembryonic antigen (CEA) promoter (Cp-CDglyTK) on colorectal carcinoma xenograft in nude mice.

The plasmid pcDNA3.1 (-) Cp-CDglyTK was transfected into the CEA-positive SW480 and CEA-negative HeLa cells respectively.The expression of suicide gene was detected by RT-PCR.And the transfected cells were treated with 5-fluorocytosine (5-FC) and ganciclovir (GCV) at different concentrations and the cell-killing and bystander effects assayed by methyl thiazolyl tetrazolium (MTT).By a transplantation of cultivated cells,SW480 or HeLa cell lines were injected subcutaneously into right axillary of nude mice to establish 96 SW480 and 72 HeLa tumor animal models.Nude mice were completely randomized with statistical software according to tumor volume.For prodrug therapy, 48 SW480-bearing mice were divided equally into 4 groups ofⅠ-Ⅳ. At the same time, 48 HeLa-bearing mice were divided equally into 4 groups of Ⅴ-Ⅷ. GroupsⅠ&Ⅴ received an intratumoral injection of PBS, groups Ⅱ & Ⅵ GCV and 5-FC intratumorally, groups Ⅲ & Ⅶ PBS intraperitoneally and groups Ⅳ & Ⅷ GCV and 5-FC intraperitoneally.Forty-eight SW480-bearing mice were divided equally into 4 groups of Ⅸ~Ⅻ and 24 Helabearing ones into groups of Ⅷ & Ⅹ in therapy experiment by suicide gene plus prodrug.Six groups received an intratumoral injection of liposome Lipofectamine^(R) and plasmid CP-CDglyTK and then an intraperitoneal injection of drug.The groups of Ⅸ and Ⅷ received an injection of PBS, group XGCV, group Ⅺ 5-FC and groups Ⅻ & Ⅹ GCV and 5-FC.The observation parameters included tumor bulk, tumor weight, survival time and treatment effect in each group.

SW480 cells transfected by plasmid pcDNA3.1 (-) Cp-CDglyTK expressed CDglyTK gene.The inhibition rates of GCV and 5-FC were significantly higher than those of HeLa cells (59.87%±0.21% vs 9.90%±0.09%, P<0.01).And higher inhibition rates and stronger bystander effect existed in double versus single produg (all P<0.05).Tumor size, final tumor weight and survival time of nude mice in groups of Ⅱ,Ⅳ,Ⅵ & Ⅷ had no significant difference with groups ofⅠ,Ⅲ,Ⅴ & Ⅶ (all P<0.05).Final tumor size and weight of group Ⅻ was significantly smaller than those of groups of Ⅸ,Ⅹ and Ⅺ ((150.0±3.2) vs (522.5±1.9) and (256.8±10.4) and (260.7±2.2) mm³, (54.1±10.4) vs (682.0±12.0) and (251.8±15.1) and (271.6±17.7) mg, all P<0.05).Meanwhile, the tumor inhibition rate and survival time of group Ⅻ (92.1% and (25.7±0.8) d) were significant higher and longer than group Ⅹ (63.1% and (21.8±0.5) d) and groupⅪ (60.2% and (18.0±0.9) d)(all P<0.05).However, no significant difference existed in tumor size, final tumor weight and survival time between groups Ⅷ and Ⅹ (all P>0.05).The inhibition rate of group Ⅹ was merely 0.9%.

CDglyTK double suicide gene system driven by CEA promoter may inhibit CEA positive colorectal cancer xenograft in prodrug-treated nude mice.

To explore the expression and effect of Toll-like receptor 4 (TLR-4) in the lung tissue of rats established by passive smoking or intratracheal instillation of lipopolysaccharide (LPS).

Eight-week-old male Sprague-Dawley rats (n=15) were randomly divided into 3 groups,including: (1) group A:conventional breeding; (2) group B:the rats were placed into a 120-L organic glass box with twice-daily exposure to cigarette smoking plus an intratracheal instillation of water at Day 1 and 14; (3) Group C: exposure to cigarette smoking the same as group B plus intratracheal instillation of lipopolysaccharide (1 mg/kg) at Day 1 and 14.Four weeks later, general status, arterial blood gas, pulmonary function and histopathology were analyzed.The expressions of T LR-4 and nuclear factor kappa-B (NF-κB) were determined by immunohistochemistry.Western blot was used to measure the protein contents of T LR-4, NF-κB, p-Iκ-Kα/β, Iκ-Kα/β, IκB-α.And realtime polymerase chain reaction (PCR) was employed to detect the mRNA levels of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6).

Rats in Groups B and C were marantic with intermittent cough and dyspnea.Peak expiratory flow (PEF) and 50% expiratory flow-volume (EP50) were much lower in Group C ((10.6±1.4), (0.77±0.14) ml/s) than that in Groups A ((13.5±2.0), (1.01±0.08) and B (12.3±0.9), (0.91±0.10) ml/s)(all P<0.05).Accumulated volume (AV) and carbon dioxide pressure (PCO2) were much higher in Groups B ((4358±1501) ml, (52.77±1.97) mm Hg)(1 mm Hg=0.133 k Pa) and C ((10 077±1866) ml, (51.03±4.96) mm Hg) than that in Group A ((1735±798) ml, (39.57±1.43) mm Hg)(all P<0.05).Hematoxylin and eosin stain showed chronic bronchitis and emphysema in Groups B and C.Besides, quantitative analysis demonstrated that in unit area, mean lining interval (MLI) and destruction index (DI) in Group B ((84±13) μm, 0.228±0.047) and Group C ((86±10) μm, 0.294±0.060) significantly increased versus Group A ((65±6) μm,0.036±0.012)(all P<0.05).Immunohistochemical staining indicated that the expression of TLR-4 in cytoplasm and cytomembrane and NF-κB in nucleus markedly increased in Groups B and C versus Group A.Relative expressions of TLR-4 and NF-κB assayed by Western blot increased in Group B (0.68±0.03, 0.21±0.08) and Group C (1.12±0.11, 0.59±0.06) than that in Group A (1.36±0.07, 1.04±0.08).Compared with Group A, the expression levels of T LR-4, NF-κB and IκB-αand the phosphorylation levels of Iκ-Kα/βin Group B and C significantly increased (all P<0.05).The mRNA levels of TNF-αand IL-6 increased in Group B (3.95±0.29, 5.04±0.28) and C (5.33±0.26, 7.23±0.39) versus that in Group C (1.00±0.37, 1.00±0.25)(all P<0.05).

Both passive smoking and intratracheal instillation of LPS may cause lung injury analogous to chronic obstructive pulmonary disease via NF-κB signaling pathway.And TLR4 plays an important role in this process.

To explore the effects of glycyrrhizin on airway mucus hypersecretion induced by interleukin-13 (IL-13) in rats.

A total of 50 SD rats were divided randomly into 5 groups with a random digit table:control group, IL-13 group, and different dosage (25, 50, 75 mg/kg) glycyrrhizin groups.The integral of expression intensity in positive cells of airway epithelium under mucus histochemical stain was calculated with modality-quantitative method.HBE-16 cells were divided into 6 groups: negative control (physiological saline), IL-13 control (10μg/L IL-13+ physiological saline), different concentration glycyrrhizin interference (10μg/L IL-13 + 25, 50 and 75μmol/L glycyrrhizin, respectively) and positive control (10μg/L IL-13+ 25μmol/L zopolrestat).The expression of mucin (MUC) 5AC mRNA, MUC5AC protein, aldose reductase (AR) activity and reactive oxygen species (ROS) content were detected by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, fluorometric method and fluorescence intensity with General Oxidative Stress Indicator (CM-H₂DFDA) catheter respectively.

In vivo, the integral of expression intensity in positive stain cells of airway epithelium were 0.12±0.03,0.87±0.13,0.56±0.08,0.46±0.06 and 0.35±0.04 respectively while the integral of different dosage glycyrrhizin groups was significantly lower than that of IL-13 group (all P<0.05) with dose depentency and the IL-13 group was stronger than control group (P<0.05).In vitro, the index of AR activity and ROSat 48 h of HBE16 cells in every group were 0.156±0.021,0.692±0.039,0.436±0.019,0.323±0.042 and 0.290±0.027; 5.127±0.033, 24.257±3.263, 11.966±0.283, 8.892±0.521 and 6.426±0.173 respectively.The indices of IL-13 control group were higher than those of negative control group (P<0.05) while those of different concentration glycyrrhizin interference groups were lower than those of IL-13 control group (all P<0.05).The expressions of MUC5AC mRNA and protein of HBE16 cells in every group were 0.82±0.05, 3.22±0.12, 2.57±0.34, 2.09±0.54 and 1.58±0.22; 0.18±0.04, 0.65±0.15, 0.48±0.11,0.33±0.19 and 0.26±0.06 respectively.The indices of IL-13 control group were higher than those of negative control group (P<0.05) and those of different concentration glycyrrhizin interference groups were lower than those of IL-13 control group (P<0.05).

Glycyrrhizin may inhibit the expression of MUC5AC mRNA and MUC5AC protein induced by IL-13 and control the hypersecretion of airway mucus.

To explore the efficacy of muscovite on iodoacetamide-induced ulcerative colitis in rats and elucidate its possible mechanism.

Ulcerative colitis was induced in female Sprague-Dawley (SD) rats by an intracolonic injection of iodoacetamide.A total of 48 rats were divided randomly (by the method of random digits table) into 6 groups:control group, model group, low-dose muscovite group (360 mg/kg), high-dose muscovite group (720 mg/kg), 5-aminosalicylie acid (5-ASA) group and muscovite plus 5-ASA group (combined treatment), and each group had 8 rats.The body weight, disease activity index (DAI), macroscopic damage and microscopic score of rats in each group were subsequently evaluated after dosing for 7 days.The protein levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8) and myeloperoxidase (MPO) activity were detected by enzyme-linked immunosorbent assay (ELISA) while the activity of nuclear facor (NF) -κB was determined by immunohistochemistry.One way ANOVA and rank-sum test were used.

After doing, body weight macroscopic damage, microscopic score, TNF-αconcentration, MPO and NF-κB activity of rats in each group were all significantly correlated with the dose of muscovite (r=0.573,-0.647,-0.569,-0.681,-0.811,-0.842, all P<0.05).High-dose muscovite group had no significant difference with 5-ASA group in body weightt, DAI, macroscopic damage, microscopic score, IL-8 concentration, TNF-αconcentration, MPO and NF-κB activities ((166±5) vs (167±5) g, 0.33 (0.00, 1.17) vs 0.17 (0.00, 0.83), 2.50 (2.00, 4.00) vs 3.00 (2.00, 3.00),3.00 (2.00,3.00) vs 2.50 (2.00,3.00), (109±17) vs (111±15) pg/ml, (166±38) vs (155±45) pg/ml, (52±6) vs (49±4) U/g, 7.39±0.42 vs 7.41±0.34, all P>0.05).The MPO and NF-κB activities of combined treatment group were lower than those of 5-ASA group ((40±4) vs (49±4) U/g, 4.67±0.72 vs 7.41±0.34, all P<0.05).However, other indices showed no significant difference with 5-ASA group (all P>0.05).

Rectal administration of muscovite ameliorates colonic inflammation of iodoacetamide-induced colitis.Its underlying mechanism is probably due to the regulation of inflammatory response.Muscovite may be a potential therapeutic agent for treatment of ulcerative colitis.

To establish a method of detecting circulating immunoglobulin E (IgE) autoantibodies for BP180NC16A and evaluate its significance in bullous pemphigoid (BP).

GST-NC16A fusion proteins were expressed in Escherichia coli using the pGEX-2T expression system and purified by glutathione affinity chromatography.For optimal working conditions of enzyme-linked immunoabsorbent assay (ELISA),checkerboard titrations were performed with serial dilutions of antigen.Also optimized dilution of secondary antibody was confirmed.Sera samples from 56 patients with BP, 24 healthy controls,18 with pemphigus and 1 with Stevens-Johnson syndrome at our hospital during February 2011 to October 2012 were examined by the modified ELISA approach.The optimal cut-off point for a positive result was selected with receiver operating characteristic analysis.

The optimized ELISA was performed with plates coated with 500μg/ml GST-NC16A.And the optimal dilutions of sera samples and secondary antibody were 1∶10 and 1∶1000 respectively.According to the established cut-off value (0.549),40 of 56 BP patients and none of controls had detectable levels of BP180NC16A IgE.

The established ELISA provides a highly specific tool for the detection of IgE anti-BP180NC16A in BP patients.