Objective:To establish mice models of Staphylococcus aureus bloodstream infections so as to investigate the inflammatory responses and histopathological changes in bloodstream infections (BSIs) mice.Methods:C57BL/6 mice were inoculated with S.aureus intravenously or intraperitoneally to induce BSIs.Survival rate , weight loss and murine sepsis scores ( MSS ) were ob-served.Blood samples and tissue homogenates were plated on agar to determine bacterial burden .Inflammatory proteins ( CRP,PCT) and cytokines ( IL-1β, IL-6 and TNF-α) were determined by ELISA kits.Histopathologic changes were also assessed by pathological inflammation scores(PIS),macroscopic and microscopic examination.Results: About 70% survival rate was observed in 4.5×108 CFU/ml S.aureus induced BSIs mice.Body weight decreased and sepsis scores increased significantly since 24 h post-infection in BSIs mice,and more prominent in IV group.The counts of WBC began to significantly increase at 3 h post-infection,while CRP and PCT levels peaked at 48 hours in IV and IP groups ( 60.80 ±5.63 vs 40.58 ±7.54 for CRP;6.796 ±1.16 vs 2.740 ±0.36 for PCT ) . Moreover,the levels of IL-1β,IL-6 and TNF-αin serum and tissue homogenates ( liver,lungs,and kidneys ) were significantly elevated in BSIs mice.Pathological changes in tissues (liver,lungs and kidneys) and higher pathological inflammation scores (PIS) were also observed in BSIs mice.Conclusion:Our study represents an effective approach for S.aureus BSIs model to mimic human sepsis.Our results demonstrated that inflammation protein (PCT,CRP) and cytokines(IL-6,IL-1βand TNF-α) play an important role in the in-flammatory response and histopathological changes during BSIs caused by S .aureus.

Objective To establish mild cognitive dysfunction (MCI) models in elderly rats,and to investigate the pathophysiological features.Methods Totally 40 SD rats (14 to 18-month-old) were randomly divided into 2 groups:the model group (n=20) and the sham operation group (n=20).Bilateral carotid artery stenosis was prepared in the model group while bilateral carotid artery was seperated with no bilateral narrowing in the sham operation group.30 days after the operation,Morris water maze test was performed,pathomorphological and electron microscopic observations of the cerebral tissue were examined and the expression of G protein-coupled receptor kinase 2(GRK2) in hippocampus tissue w detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blottin.Results The mortality in model group was only 10％.Pathological morphology and ultrastructure showed that hippocampal tissue structure was almost normal in sham operated group,but in model group group,hippocampal CA1 pyramidal cells were in ischemic demyelination,arranged loose,and part of the cells showed nucleus pyknosis,deeply stained; there was no obvious infarct in white matter,part of the white matter fiher hecame thinner and disorder,nucleolus became smaller and steped aside,cytoplasmic electron density increased,lipofuscin appeared occasionally.Rough endoplasmic reticulum and Golgi were expanded,cytosolic free ribosomes increased,part of mitochondria became swelled,vacuolated.Morris water maze test results showed that the average escape latency in model group was longer than in sham group (P＜0.05).In spatial probe test,the average time of crossing the first original platform in model rats was significantly longer than the sham operated group [(36.80±7.68) s vs.(20.87±6.16)s,P＜0.05].The average number of crossing the original platform in 60 seconds in model group was significantly less than in sham group(1.43±0.51 vs.3.10±1.45,P＜0.05).The expressiones of GRK2 mRNA and protein in the hippocampus were significantly increased in model group rats than in sham group (P＜0.05).Conclusions The model of severe CCA stenosis in elderly rats can be applied for MCI animal models with good stability and repeatability.Compared with sham group,the cells morphology and ultrastructure in model group appeare more obvious pathological changes and mild impairments in cognitive function.GRK2 may play an important role in the development of MCI.

Objective To investigate the clinical significance of the serum sialic acid (SA) detection for the diagnosis and therapy monitoring in liver cancer patients.Methods Patients and healthy people of Chinese academy of medical science cancer hospital from January 2011 to October 2012,were enrolled,including 221 liver cancer patients (183 primary hepatic carcinoma patients and 38 metastatic hepatic carcinoma patients),117 benign liver disease patients and 150 healthy people.The concentration of serum SA were tested by ROCHE P800.The intra-assay and inter-assay coefficient of variation (CV) of SA kit were evaluated by use of low and high concentration samples,measured for 5 days and 4 times each day.Receiver operating characteristic (ROC) curve were used to determine the cut-off of SA using data of 183 cases of primary liver cancer and 150 healthy controls.The area under the curve (ROC-AUC) were used to evaluate the diagnostic value of SA.The changes of serum SA level in 103 cases of primary hepatic carcinoma patients were monitored before therapy and at the 1 st day,7 th day,14 th day,1 st month,3rd month,6 th month and 9 th month after treatment.SPSS16.0 was used to analyse the results.Results The intra and inter-day CVs for low level sample were 2.4％ and 3.2％ respectively,and for high level sample were 2.2％ and 3.1％.The cut-off value of the serum SA was 659 mg/L for liver cancer,the sensitivity and specificity was 63.4％ (1 16/183) and 94.7％ (142/150) respectively.The serum SA level of liver cancer group [(726 ± 173) mg/L] was higher than that of liver benign disease patients group [(552 ± 128) mg/ L] and healthy controls group [(599 ± 62) mg/L,U values were 1832.52 and 887.00,P ＜ 0.01].The serum SA level were tracked in 103 cases of primary hepatic carcinoma patients during therapy period.The serum level of SA elevated to [(817 ± 193) mg/L,t =-3.272,P ＜ 0.05] at I st week after treatment and kept at high level until late in 1st month after treatment [(782 ±173) mg/L,t =-2.694,P＜0.05].In the 3rd month,the SA level decreased to that of pretreatment [(662 ± 138) mg/L,t =1.225,P ＞ 0.05].In the 6th months,the SA level declined to [(615 ± 144) mg/L,t =1.999,P ＜0.05],as well as the level of healthy control group.There were 85 cases of hepatic carcinoma patients with decreased SA level compared with that of pretreatment,and the coincidence rate was 82.5％ (85/103),the Kappa value was 0.79.There were 5 cases of patients with hepatic carcinoma relapse after treatment in 9 th months and the SA levels increased significantly to (939 ± 175) mg/L.Conclusion The serum SA has significant values possibly in the diagnosis and therapy monitoring in liver cancer patients.

Objective To investigate the risk factors of ventilator-associated pneumonia (VAP) and the distribution and drug resistance of pathogens in intensive care unit (ICU) of county hospital. Methods 234 patients on mechanical ventilation for more than 48 hours admitted to ICU of Shexian People's Hospital of Huangshan City from January 2016 to June 2018 were enrolled. The clinical data of all patients including gender, age, past medical history, exposure to antibiotics, medication, the duration of mechanical ventilation, the length of ICU stay, serum albumin, tracheotomy, re-intubation, prognosis, and pathogenic bacteria and drug sensitivity test of VAP patients were collected. The patients were divided into VAP group and non-VAP group according to the occurrence of VAP. The differences of each index between the two groups were compared. The risk factors of VAP were analyzed by multivariate Logistic regression. The distribution and drug resistance of pathogenic bacteria in sputum culture of lower respiratory tract of VAP patients were analyzed. Results Among the 234 patients on mechanical ventilation, 95 patients had VAP, and the incidence of VAP was 40.60%. ① Risk factors of VAP: it was shown by univariate analysis that there were significant differences between VAP patients and non-VAP patients in past history, the duration of mechanical ventilation, the length of ICU stay, albumin < 28 g/L, antibiotic exposure and tracheotomy, but there were no significant differences in gender, age, glucocorticoid, sedative, gastric motility and coma between the two groups. It was shown by multivariate Logistic regression analysis that brain injury and cerebrovascular accident, the duration of mechanical ventilation > 7 days, albumin < 28 g/L and tracheotomy were independent risk factors for VAP occurrence [brain injury: odds ratio (OR) =41.40, 95% confidence interval (95%CI) = 2.14-799.60, P = 0.014; cerebrovascular accident: OR = 36.07, 95%CI =1.86-699.64, P = 0.018; the duration of mechanical ventilation > 7 days: OR = 1.23, 95%CI = 1.11-1.36, P < 0.001;albumin < 28 g/L: OR = 2.27, 95%CI = 1.03-5.01, P = 0.042; tracheotomy: OR = 3.33, 95%CI = 1.30-8.56, P = 0.012].② Distribution and drug resistance of VAP pathogens: a total of 108 strains of pathogens were isolated from sputum samples of 95 patients with VAP. Gram-negative (G-) bacteria accounted for 86.11% (93/108). The isolation rate of Klebsiella pneumoniae was the highest, reaching 31.48% (34/108); the isolation rates of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii were 22.22% (24/108), 8.33% (9/108) and 9.26% (10/108), respectively. Gram-positive (G+) bacteria accounted for 6.48% (7/108), of which Staphylococcus aureus was 4.63% (5/108); and fungi was 7.41% (8/108). Drug resistance analysis showed that Klebsiella pneumoniae was 100% sensitive to amikacin (AMK), meropenem (MEM) and polymyxin (POL), and were suggested as the preferred drug. Pseudomonas aeruginosa was 100% sensitive to AMK, tobramycin (TOB) and POL, but 100% resistant to compound trimethoprim (PCST). Stenotrophomonas maltophilia was 100% sensitive to PCST and 100% resistant to AMK, piperacillin (PIP), piperacillin tazobactam (TZP) and TOB. Acinetobacter baumannii was 100% sensitive to cefoxitin (FOX), cefuroxime (CXM) and POT. Staphylococcus aureus was 100% sensitive to gentamicin (GEN), furantoin (NIT), rifampicin (RIF), vancomycin (VAN) and teicoplanin (TEC), while the drug resistance to clindamycin (CLI) and penicillin (PEN) was high (both 80.00%). Most pathogens were multidrug-resistant. The mortality of patients with multidrug resistant bacteria infection was significantly higher than that of non-multidrug resistant bacteria infection [51.85% (28/52) vs. 30.56% (11/36), χ2= 4.240, P = 0.046]. Conclusions VAP was associated with brain injury and cerebrovascular accident, duration of mechanical ventilation > 7 days, albumin < 28 g/L and tracheotomy. VAP patients were infected mainly with G- bacteria and showed multiple drug resistance.

Objective To identify disease-causing mutations in a large panel of Chinese Han patients with hereditary spastic paraplegia(HSP).Methods The coding sequence of the SPG11 gene in the probands of 28 families with ARHSP and 14 sporadic HSP patients was analyzed,and the identified changes in the sequence were tested to exclude being a benign polymorphism by sequencing 200 chromosomes from normal controls.Results Identified 13 causative mutations in SPG11 gene in 7 ARHSP and 3 sporadic HSP The mutations were:c.5977C>T/p.Q1993X、c.4668T>A/p.Y1556X、c.6898_6899delCT/p.L2300AfsX23.38、c.3719_3720delTA/p.11240VfsX263、c.733_734delAT/p.M245VfsX246、c.7088_7089insATTA/p.Y2363X、c.2163_2164insT/p.1722Yfsx731、c.7101-7102insT/p.K2368X、c.6790_6791insC/p.12264PfsX2339、c.654_655delinsG/p.S218RfsX219、c.7151+4_7151+7delAGTA/p.K2384fsX2386、c.6355-21_6355-18delTCT、c.3004C>T/p.G1002X.Among them,12 were novel mutations.The rate of mutation in the SPG11 gene was 25.0%(7/28)in ARHSP,6/6 in ARHSP-TCC and 3/14 in sporadic cases.Conclusions In Chinese Han population,patients with ARHSP-TCC and sporadic HSP-TCC should be screened for SPG11.Sequencing of the SPG11 gene in these patients with is valuable for clinical diagnostic testing.

BACKGROUND/AIMS: This study was to investigate whether transforming growth factor-β1 (TGF-β1) plays a role in hyperalgesia in chronic pancreatitis (CP) and the underlying mechanisms. METHODS: CP was induced in male adult rats by intraductal injection of trinitrobenzene sulfonic acid (TNBS). Abdominal hyperalgesia was assessed by referred somatic behaviors to mechanical stimulation of rat abdomen. Dil dye injected into the pancreas was used to label pancreas-specific dorsal root ganglion (DRG) neurons. Whole cell patch clamp recordings and calcium imaging were performed to examine the effect of TGF-β1 on acutely isolated pancreas-specific DRG neurons. Western blot analysis was carried out to measure the expression of TGF-β1 and its receptors. RESULTS: TNBS injection significantly upregulated expression of TGF-β1 in the pancreas and DRGs, and TGF-β1 receptors in DRGs (T9-T13) in CP rats. Intrathecal injection of TGF-β receptor I antagonist SB431542 attenuated abdominal hyperalgesia in CP rats. TGF-β1 application depolarized the membrane potential and caused firing activity of DRG neurons. TGF-β1 application also reduced rheobase, hyperpolarized action potential threshold, and increased numbers of action potentials evoked by current injection of pancreas-specific DRG neurons. TGF-β1 application also increased the concentration of intracellular calcium of DRG neurons, which was inhibited by SB431542. Furthermore, intrathecal injection of TGF-β1 produced abdominal hyperalgesia in healthy rats. CONCLUSIONS: These results suggest that TGF-β1 enhances neuronal excitability and increases the concentration of intracellular calcium. TGF-β1 and its receptors are involved in abdominal hyperalgesia in CP. This and future study might identify a potentially novel target for the treatment of abdominal pain in CP.
Abdomen ; Abdominal Pain ; Action Potentials ; Adult ; Animals ; Blotting, Western ; Calcium ; Diagnosis-Related Groups ; Fires ; Ganglia, Spinal ; Humans ; Hyperalgesia ; Injections, Spinal ; Male ; Membrane Potentials ; Neurons* ; Pancreas ; Pancreatitis, Chronic* ; Rats*

Objective: To detect the levels of D-dimer (D-D) and fibrinogen (Fbg) in plasma of patients with malignant tumors, and to evaluate the diagnostic value of combined detection of D-D and Fbg. Methods: The clinical data of 99 patients with malignant tumors were retrospectively analyzed, including 47 patients in DVT group and 52 patients in control group. DVT diagnosis was based on color Doppler or venography. The ROC curve was used to evaluate the diagnostic value of D-D, Fbg and the combination of D-D and Fbg for the malignant tumor patients with DVT. Results: The D-D levels in the DVT group and the control group were 7.17 μg/ml and 4.25 μg/ml, respectively, and the Fbg levels were 4.01 mg/ml and 2.02 mg/ml, respectively, and the differences were statistically significant (all P<0.01). For the area under the ROC curve (AUC), specificity and sensitivity for diagnosing malignant tumors with DVT, D-D were 0.728, 62.50%, and 72.50%, respectively, and Fbg was 0.642, 65.50%, and 58.80%, respectively, and D-D+Fbg was 0.764, 83.30% and 59.00% respectively. Conclusions D-D and Fbg has a significantly high level in malignant tumor patients with DVT. The combined detection of D-D and Fbg has potential application value in the early diagnosis, early intervention and better prognosis for the malignant tumors patients with DVT.

Objective:According to the characteristics and common problems of hematology analysis in cancer patients, an autoverification scheme for cancer patients was formed, and the effectiveness and efficiency of the autoverification scheme were verified.Methods:The hematology review of international consensus and ourselves were respectively combined with Chinese multicenter autoverification rules to form two autoverification schemes. 10 063 blood samples (460 cases reviewed by microscope) were selected as the establishment group. Retrospective judgment was made in the instrument middleware, and various indexes such as autoverification pass rate and missed detection rate of different schemes were compared. By analyzing the data of missed cases one by one, the autoverification rules are adjusted according to the characteristics of diagnosis and treatment of cancer patients. By analyzing the platelet count variation range within 7 days in 19 300 cases, the Delta rules of platelet count were established. The platelet count Delta rules and the adjusted autoverification rules were combined to form the autoverification rules of our hospital and then combined with our hematology review rules to form the autoverification scheme of our hospital. The establishment group and verification group (10 876 cases, including 1 740 cases of microscopic examination) of the autoverification schemes were judged. The recognition function of Ethylenediaminetetraacetic acid-dependent pseudo thrombocytopenia (EDTA-PTCP) and PLT Delta check were programmed in the laboratory information system (LIS), and other rule judgment functions are performed in middleware. After four months of clinical trial application of 61 602 specimens, the effectiveness of our autoverification scheme was comprehensively evaluated.Results:The autoverification pass rates of international hematology review rules, our review rules, and Chinese multicenter autoverification rules are 46.36%, 52.26%, and the missed detection rates are 2.02%, 1.06%, respectively. The autoverification pass rates of our hospital autoverification scheme in the establishment group and the verification group are 51.19% and 52.78%, the missed detection rates are 0% and 0.03%, and the true positive rate are 100% and 99.95%, respectively. 56.06% of cases were passed automatically during the clinical trial application, and there were no missing cases, the true positive rate is 100%. The performance of our autoverification scheme is superior to the current autoverification schemes combined with mainstream hematology review rules and autoverification rules. The median time of TAT by autoverification was shortened by 15 minutes, and the 90th percentile time was shortened by 58 minutes, which was significantly lower than that of the same period last year. The marker function of "EDTA-PTCP" identified 31 special patients and 68 samples had been analyzed in total. After correction, the median increase of PLT was 76.5×10 9/L ( Z=-7.17, P<0.001). Conclusions:This study has established an autoverification scheme that combined by rules of hematology review and autoverification rules. It is suitable for cancer patients with high pass rate and very low rate of missed detection. This autoverification scheme can ensure the accuracy of the hematology analysis of cancer patients in our hospital and improve work efficiency.

Objective:To investigate the different outcomes of two types of acute kidney injury (AKI) according to standard of Kidney Disease: Improving Global Outcomes-AKI (KDIGO-AKI), and to analyze the risk factors that affect the prognosis of intensive care unit (ICU) patients in China.Methods:A secondary analysis was performed on the database of a previous study conducted by China Critical Care Clinical Trial Group (CCCCTG), which was a multicenter prospective study involving 3 063 patients in 22 tertiary ICUs in 19 provinces and autonomous regions of China. The demographic data, scores reflecting severity of illness, laboratory findings, intervention during ICU stay were extracted. All patients were divided into pure AKI (PAKI) and acute on chronic kidney disease (AoCKD). PAKI was defined as meeting the serum creatinine (SCr) standard of KDIGO-AKI (KDIGO-AKI SCr) and the estimated glomerular filtration rate (eGFR) at baseline was ≥ 60 mL·min -1·1.73 m -2, and AoCKD was defined as meeting the KDIGO-AKI SCr standard and baseline eGFR was 15-59 mL·min -1·1.73 m -2. All-cause mortality in ICU within 28 days was the primary outcome, while the length of ICU stay and renal replacement therapy (RRT) were the secondary outcome. The differences in baseline data and outcomes between the two groups were compared. The cumulative survival rate of ICU within 28 days was analyzed by Kaplan-Meier survival curve, and the risk factors of ICU death within 28 days were screened by Cox multivariate analysis. Results:Of the 3 063 patients, 1 042 were enrolled, 345 with AKI, 697 without AKI. The AKI incidence was 33.11%, while ICU mortality within 28 days of AKI patients was 13.91% (48/345). Compared with PAKI patients ( n = 322), AoCKD patients ( n = 23) were older [years old: 74 (59, 77) vs. 58 (41, 72)] and more critical when entering ICU [acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score: 23 (19, 27) vs. 15 (11, 22)], had worse basic renal function [eGFR (mL·min -1·1.73 m -2): 49 (38, 54) vs. 115 (94, 136)], more basic complications [Charlson comorbidity index (CCI): 3 (2, 4) vs. 0 (0, 1)] and higher SCr during ICU stay [peak SCr for diagnosis of AKI (μmol/L): 412 (280, 515) vs. 176 (124, 340), all P < 0.01]. The mortality and RRT incidence within 28 days in ICU of AoCKD patients were significantly higher than those of PAKI patients [39.13% (9/23) vs. 12.11% (39/322), 26.09% (6/23) vs. 4.04% (13/322), both P < 0.01], while no significant difference was found in the length of ICU stay. Kaplan-Meier survival curve analysis showed that the 28-day cumulative survival rate in ICU in AoCKD patients was significantly lower than PAKI patients (Log-Rank: χ2 = 5.939, P = 0.015). Multivariate Cox regression analysis showed that admission to ICU due to respiratory failure [hazard ratio ( HR) = 4.458, 95% confidence interval (95% CI) was 1.141-17.413, P = 0.032], vasoactive agents treatment in ICU ( HR = 5.181, 95% CI was 2.033-13.199, P = 0.001), and AoCKD ( HR = 5.377, 95% CI was 1.303-22.186, P = 0.020) were independent risk factors for ICU death within 28 days. Conclusion:Further detailed classification (PAKI, AoCKD) based on KDIGO-AKI SCr standard combined with eGFR is related to ICU mortality in critical patients within 28 days.