Volume 93  ·   Issue 10  ·  2013

To explore the characteristics, early symptoms and valuable experiences in the diagnosis and treatment of urine-induced sepsis.

The clinical data of 30 cases of urineinduced sepsis between August 2008 and April 2012 were reviewed retrospectively.Among them, YAG laser lithotripsy (n=3) and transurethral prostatic resection (TURP)(n=2) were performed.One case was after transurethral resection of bladder (TURB).There were 3 cases after cystectomy and Bricker and another 18 during and after percutaneous nephrolithotomy (PCNL).

A total of 12 PCNL cases have significant drop in blood pressure during operation with non-bleed causes.A total of 26 cases developed symptoms of chills and high fever after 2 hours of surgery.The earliest symptoms of urine-induced sepsis were decline in blood pressure and high fever.After resistance to infection and symptomatic treatment actively, 27 patients were rescued successful (90%), 3 patients died (10%).Among the 3 deceased patients, 1 case was after YAG laser lithotripsy under ureteroscope, 1 case was after PCNL, 1 case was after TURP.

The onset of urine-induced sepsis onset may be insidious.But this condition deteriorates rapidly and it is rather difficult to reverse.Most cases could be managed when the onset characteristics were recognized early.Prevention and early treatment are keys of lowering the mortality.

To explore the value of detecting mutations on epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer (NSCLC) tissue by TaqMan-amplification refractory mutation system (TaqMan-ARMS).

TaqMan-ARMS and DNA sequencing were used to detect the EGFR exon 19 and 21 mutations in tumor tissues and the samples collected from 199 patients at 4 different 3 A hospitals in Beijing from January 2008 to March 2011.

The rate of mutations in EGFR exon 19 and 21 was 19.1% (38/199), according to their different pathological types.Based upon TaqMan-ARMS, the classification was as followed: adenocarcinoma (35.0% (36/103)), squamous carcinoma (2.2% (2/93)) and adenosquamous carcinoma (0).According to DNA sequencing, they were 19.6% (39/199), 35.9% (37/103), 2.2% (2/93) and 0 respectively.Thus, no statistically significant difference existed between two Methods (McNemar Test, P=1.000,κ=0.984).The mutation rate of adenocarcinoma was higher than those of squamous and adenosquamous carcinoma.

The detection of EGFR mutations is highly consistent in the NSCLC tissue by the Methods of TaqMan-ARMS and DNA sequencing.

To explore the expression of costimulatory molecule CD40 in thyroid tissue of Graves′disease patients and understand its immune pathogenetic significance.

From January 2008 to December 2011,8 patients undergoing partial thyroidectomy for Graves′disease (n=3) or non-toxic goiter (n=5) at Affiliated Suzhou Hospital of Nanjing Medical University and Third Affiliated Hospital of Soochow University were enrolled.Using the method of immunohistochemistry, the expression of CD40 was detected in their thyroid tissues.Variation in CD40 expression on thyroid follicular (TFC) in primary cultures was analyzed in the absence (no stimulation group) or presence of inflammatory cytokines including interleukin interferon-γ (IFN-γ)(IFN-γstimulation group), interferon-6 (IL-6)(IL-6 stimulation group) and tumor necrosis factor (TNF)-α (TNF-α stimulation group) with flow cytometry.IFN-γ-stimulated TFC were cultured with agonist CD40 monoclonal antibody (5C11)(IFN-γ+ CD40 group) or isotypic mouse IgG (mIgG) antibody (IFN-γ+mIgG group).And the proliferation of TFC was assessed by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assays for each donor.The production of free triiodothyronine (FT3) and free thyroxine (FT4) and the release of thyroglobulin (Tg) were measured with radioimmunoassays.

The expression of CD40 on infiltrated lymphocytes and TFC were detected in Graves′disease but not in non-toxic goiter patient tissues.Compared with no stimulation group (23.7%±7.3%), the expression of CD40 on TFC increased in IFN-γ stimulation group (86.4%±4.6%), IL-6 stimulation group (90.0%±4.2%) and TNF-α stimulation group (87.3%±4.2%).Compared with the IFN-γ+mIgG group (0.75±0.06), the TFC proliferation of IFN-γ+CD40 group (1.14±0.14) significantly increased (P<0.01).The levels of FT3, FT4 and Tg secretion of IFN-γ+CD40 group were (1.10±0.15) pmol/L, (0.80±0.14) pmol/L and (30.23±1.60) μg/L respectively.They were all significantly increased compared with the IFN-γ+ mIgG group,of which the FT3,FT4 and Tg production were (0.76±0.07) pmol/L, (0.63±0.09) pmol/L and (21.37±3.22) μg/L respectively (all P<0.05).

CD40 is abnormally expressed in thyroid tissue of Graves′disease patients.And its costimulatory signal may take part in the immunopathologic mechanism of Graves′disease.

To explore the values of arginine stimulation, oral glucose-insulin release and intravenous glucose tolerance tests to assess the function of islet β-cell in individuals with differential glucose tolerance.

Healthy subjects (n=38) and those with impaired glucose regulation (IGR) without a family history of diabetes (n=31), 32 normal glucose tolerance (NGT,n=32) and impaired glucose regulation as first-degree relatives of type 2 diabetes mellitus (DM) (IGR, n=36) and 35 newly-diagnosed type 2 DM (n=35) were recruited.All of them received arginine Cpeptide releasing test (AST), oral glucose-insulin release test (OG-IRT) and intravenous glucose tolerance test (IVGTT).ACR_ARGwas used to reflect non-glucose stimulated insulin secretion function, AIR_0-10for first-phase insulin secretion function and ΔI₃₀/ΔG₃₀for early insulin secretion function.The changes of islet-β-cell function indicators were detected in individuals with different glucose tolerance.

①No significant differences existed in ACRARG among five groups (all P>0.05).②In terms of AIR_0-10, the patients of type 2 DM had lower levels than those with IGR and NGT with or without a family history of DM (all P<0.01).No significant difference existed among the subjects of IGR regardless of a family history of DM (P>0.05).However, the subjects of IGR with or without a family history of DM had lower levels than those with NGT regardless of a family history of DM (P<0.01 or P<0.05).The subjects of NGT with a family history of DM had lower levels than those with NGT without a family history of DM (50.0 (24.3- 85.1) vs 69.4 (46.1-94.8),P<0.05).③ΔI₃₀/ΔG₃₀: no significant difference existed between the patients of type 2 DM and IGT with a family history of DM (P>0.05).However, the subjects of type 2 DM had lower levels than those of IGR without a family history of DM and NGT with or without a family history of DM (all P<0.01).The subjects of IGR with or without a family history of DM had lower levels than those with NGT regardless of a family history of DM (all P<0.01).No significant difference existed among the subjects of NGT with or without a family history of DM (P>0.05).And it was the same with IGR group.

The acute C-peptide response to arginine stimulation test may not be used to evaluate the early phase insulin secretion among the subjects of earlystage DM, impaired glucose regulation and high diabetic risks.

To explore the clinical application of transport distraction osteogenesis arthroplasty (TDOAP) in the treatment of temporomandibular joint (TMJ) bony ankylosis.

From December 1999 to December 2011, a total of 73 patients (89 sides of TMJ) underwent TDOAP were included.There were 39 males and 34 females with a mean age of 19.6 years (range:3-60).Among them, 27 patients were recurrence cases and 30 cases were accompanied with micrognathia and obstructive sleep apnea-hypopnea syndrome (OSAHS).The mean preoperative degree of mouth opening was 6.6 mm (range:1-20).After a release of ankylosis, a transport disc was performed and fixed to ramus with a distractor.Distraction began at Days 4-8 postoperation.The distraction rhythm and rate were 0.25 mm four times daily.Distraction stopped when planning distance was achieved.And a distractor was maintained in place for 3-6 months after completion of distraction and then removed.Active postoperative training of mouth opening was implemented.

The mean distance of distraction was 15.3 mm (range:12-23).The range of mouth opening of 65 patients increased to normal and bone formation in gaps were perfect.The mean follow-up period was 44.8 months (range:18-102).Eight cases were recurrent.And 1/37 over 15 years old and 7/36 under 15 years old had reankylosis.

TDOAP is an effective treatment for TMJ ankylosis.A pediatric patient,especially recurrent, should be operated after adolescence to decrease reankylosis.Micrognathia and OSA HS should be also considered during the treatment of ankylosis.

To explore the prevalence and clinical characteristics of isocitrate dehydrogenase (IDH)1 R132 and IDH2 R140/R172 gene mutations in acute myeloid leukemia (AML) patients.

Polymerase chain reaction (PCR) and direct sequencing were used to sequence exon 4 of IDH gene in 570 AML patients from 2005 to 2011.

In a cohort of 570 patients, AML IDH gene mutation was found in 90 (15.79%) patients.ID H1 and IDH2 mutations were detected in 27 (4.74%) patients and 63 (11.05%) patients respectively.None of them had the combined mutations of ID H1 and ID H2.The highest frequency of IDH mutations was found in AML M1 (according to the FAB scheme) compared with all other subtypes (P<0.01).The median age was 53 years in mutated group versus 40 years in wild-type group (P=0.010).Mutated and wild-type groups had no significant difference in gender, white blood cell count at diagnosis, hemoglobin count and bone marrow blast percentage, excepting for blood platelets level (median 52×10⁹/L vs 31×10⁹/L, P<0.01).IDH gene mutations were associated with cytogenetically normal (CN)-AML, NPM1 mutations and particularly with the genotype of mutated NPM1 without FLT3-ITD.IDH gene mutations had no significant correlation with WT1, FLT3-TKD and MLL-PTD mutations.IDH mutated patients had a lower complete remission rate than unmutated in non-M3 patients (58.1% vs 77.9%,P<0.05).And the patients with mutant IDH gene were associated with a shorter overall survival (28.4% vs 51.3%, P<0.01).

IDH gene mutations are more prevalent in elder AML patients and it may constitute a molecular marker for a poor prognosis in AML.

As an activator of JNK and p38, phosphorylated MKK4 is considered to be associated with tumor progression and prognosis.This study was to examine the expression of pMKK4 and evaluate its prognostic significance in colorectal carcinoma.

A total of 343 cases of colorectal cancer were followed up to analyze the associations between the expression of pMKK4 and various clinicopathological factors.The expression of Serine257/Threonine261 pMKK4 was detected immunohistochemically by tissue microarray.

The staining of pMKK4 was present in cytoplasm of colorectal carcinoma.And the expression of pMKK4 was correlated with invasion depth (P=0.003), differentiation (P=0.018),lymph node metastasis (P<0.001), metastasis (P<0.001), hepatic metastasis (P=0.039) and TNM stage (P<0.001).The patients with strong pMKK4 staining had a better overall survival than those with lowered levels (Log rank=4.531, P=0.033).Univariate analysis indicated that the expression of pMKK4 was correlated with either overall survival (HR=0.785, P=0.035) or relapse-free survival (HR=0.788, P=0.044).In multivariate analysis, there was no prognostic significance of pMKK4 after adjusting for invasion depth,differentiation, lymph node metastasis, metastasis, liver metastasis and TNM stage.

The down-regulation of S257/T261 pMKK4 is associated with more advanced stages and it plays an important role in tumor progression.A high-level expression of pMKK4 indicates favorable clinical outcomes, but it is not an independent predictor.

To evaluate the expression of vascular endothelial growth inhibitor (VEGI) in sporadic clear cell renal cell carcinoma (CCRCC) and explore its relationships between VEGI expression, pathologic grade and tumor staging.

Western blot and immunohistochemical staining were used to detect the expression of VEGI in CCRCC cell line (786-O cells), CCRCC and paired normal kidney tissues.A total of 50 CCRCC cases were recruited.There were 37 males and 13 females with an average age of 53±12 years.The tumor sizes were<7 cm (n=33) and≥7 cm (n=17).Their pathologic grades were G1 (n=14), G2 (n=22) and G3 (n=14) and pathologic stages pT1 (n=32), 10 pT2 (n=10) and pT3 (n=8).

VEGI protein was predominantly located in cytoplasm.Compared with normal kidney tissues (mean optic density (MOD) of VEGI staining:0.40±0.16), it was lower in CCRCC tissues (MOD:0.11±0.06, P<0.01).In addition, the positive rate of VEGI expression, the expression intensity and the M OD of VEGI protein were negatively correlated with the pathologic grade of CCRCC (r=-0.640, P<0.01|r=-0.831, P<0.01| r=-0.781, P<0.01 respectively).The MOD of VEGI expression in≥7 cm tumors (MOD, 0.08±0.04) was significantly lower than that in<7 cm tumors (MOD: 0.12±0.06, P<0.05).However, there was no correlations between the VEGI protein level and age, gender and pathologic stage of patients (P>0.05).

VEGI protein is predominantly located in cytoplasm.Compared with CCRCC tissues, VEGI protein level is higher in normal ones.In consideration of negative correlations between VEGI expression, pathologic grade and tumor size,it is implied that VEGI may play a negative regulatory role in the occurrence and development of CCRCC.

To report a novel method of establishing a stable shock model of swine.

A total of 12 Guangxi BA-MA female miniature swines (aged 4-6 months) into A and B groups according to a random number table.Total body surface area (TBSA) was calculated according to the Meeb-Rubner formula.After intravenous anesthesia and other steps, solidified gasoline was used as burns material to establish TBSA 40% skin burn (Group A:45 s, Group B:50 s).Burn depth by was judged by visual and histopathological examination.Fluid resuscitation was instituted at 2 h post injury.At the same time, the hourly and pre-injury data of heart rate (H R), blood pressure (BP), urinary volume, central venous pressure (CVP) and pulmonary arterial wedge pressure (PAWP) were recorded.The actual input quantities of fluid resuscitation were assessed at the 1st and 2nd 24 h post injury.And shock condition was estimated by shock index at 2 h post injury.

All miniature swines were successfully modeled without any mortality.The burn wound observed by visual examination and light microscope showed that Group A was of deepⅡburn and Group BⅢburn.Compared with the pre-injury data, HR (111±6 vs 74±9,119±13 vs 75±13 beats/min) increased, urinary volume (0.99±0.06 vs 1.45±0.05,0.82±0.04 vs 1.47±0.06 ml·kg^-1·h^-1), CVP and PAWP decreased at 2nd 24 h post-injury in two groups (P<0.01,P<0.05).But the changes of BP (120±6/93±9 vs 120±6/94±6, 121±15/90±11 vs 124±10/91±8 mm Hg) had no statistical significance (P>0.05).The average value of HP in 48 h post-injury had no statistical significance in two groups (P>0.05).The recovery liquid quantity in the first 24 h of Group B (1.66±0.10 ml·kg^-1·1%^-1TBSA) was significantly more than Group A (1.53±0.05 ml·kg^-1·1%^-1TBSA)(P<0.05).The ratio between liquid input and output of Groups A and B showed that the comparison had no statistical significance (P>0.05).In the second 24 h post-injury, the recovery liquid quantity (0.69±0.17, 0.65±0.16 ml·kg^-1·1%^{- 1}TBSA) and the ratio between liquid in and out (1.48±0.28, 1.52±0.27) % between Groups A and B had no statistical significance (P>0.05).The shock index at 2 h post-injury showed that Groups A and B were in shock compensatory period (0.92±0.07) and mild shock state (1.00±0.16) respectively (P>0.05).

Such a shock model is stable and dependable.

To explore the effects of leflunomide active metabolite A771726 on high glucose-induced podocyte cytoskeleton and its possible signaling pathway.

The conditionally immortal human glomeroular podocytes were divided into normal glucose (NG), mannitol (MA), high glucose (HG), high glucose with PDTC (pyrrolidine dithiocarbamate, a NF-κBp65 inhibitor) and high glucose with active leflunomide metabolite A771726 groups.Western blot was used to measure the ratio of p-NF-κBp65 to total NF-κBp65.And the protein and mRNA expressions of NF-κBp65, TRPC6 and nephrin were detected by Western blot and reverse transcription polymerase chain reaction (PCR).Immunofluorescence staining was used to detect the changes in the skeleton of podocyte.

(1) Podocytes with high glucose could activate the NF-κBp65 signaling pathway.There was a significant increase of p-NF-κBp65 protein at 60 min versus 0 min (1.20±0.04 vs 0.79±0.02,P<0.01).Little activation of the pathways was observed in groups NG and MA.The up-regulated protein expression of p-NF-κBp65 induced with high glucose was significantly inhibited by PDTC and A771726 (both P<0.05).The difference of NF-κBp65 mRNA expression was not statistically significant between the groups (all P>0.05). (2) High glucose-induced podocyte activated the NF-κBp65 signaling path.Its downstream TRPC6 mRNA and protein expression significantly increased than NG while nephrin became down-regulated more than NG.PDTC and A771726 inhibited the high expression of TRPC6 while the expression of nephrin was elevated (all P<0.05). (3) Immunofluorescent assay of high glucose-induced podocyte cytoskeleton showed disorderly F-actin and a disappearance of tensile fiber after 72 h.

Active leflunomide metabolite A771726 may protect podocytes through blocking the high glucose-induced signaling pathway of NF-κBp65.

To analyze the CD40-mediated in vitro proliferation and invasiveness of lung cancer cell H1299, and explore the role of phosphatidylinositol-3-kinase-protein kinase B (PI3KAKT) signaling pathway in the above-mentioned process.

Cell counting kit-8 (CCK-8) assay was used to detect the proliferation of tumor cells.And scratch test and Transwell chambers were applied to detect the migration and invasiveness of H1299.Western blot was utilized to examine the expressions of AKT and p-AKT and enzyme-linked immunosorbent assay (ELISA) for the CD40-mediated secretion of MMP-9.

According to the CCK-8 test, the absorbance of 5C11 groups at different timepoints was significantly higher than the control group (1.72±0.18),especially when the concentration was 10 mg/L at 96 hour (2.75±0.35, P<0.05).In scratch test, the scratching width of 5C11 groups was significantly shorter than control group (1.62±0.32 mm), especially when the concentration was 10 mg/L (0.05±0.01 mm, P<0.05).Western blot showed that the level of p-AKT protein increased at 10 minutes after activating by the signal of CD40 on H1299 cells.It was higher (0.65±0.28) than the un-activating group (0.33±0.16) at 30 min (P<0.05).Upon the addition of LY294002, 5C11-mediated up-regulation of p-AKT was inhibited.In Transwell invasion assay, the number of cells on membranes in 5C11 group (168±31) were significantly higher than the control group (105±19, P<0.05) and there was no significant difference between the 5C11-added LY294002 group (118±21) and the control group (P<0.05).The expression of MMP-9 of 5C11 group (192±31 ng/L) was higher than that of the control group (110±23 ng/L, P<0.05) and no significant difference existed between the 5C11-added L Y294002 group (120±21 ng/L) and the control group (P<0.05).

The in vitro proliferation and invasiveness of lung cancer H1299 may be regulated by the CD40-mediated activation of PI3K-AKT signaling pathway.