Abstract: Objective　To understand the distribution and drug resistance of common pathogens of fungal bloodstream infection in Sichuan, and to provide reference for clinicians to empirically treat fungal bloodstream infection. Methods　From November 1, 2019 to December 31, 2020, fungal strains isolated from blood culture of patients diagnosed with bloodstream infection in 19 tertiary first-class general hospitals in Sichuan Province were collected for mass spectrometry identification and drug susceptibility, and the results were statistically analyzed, along with a retrospective analysis of clinical data. Results　A total of 255 fungal strains were received and identified by mass spectrometry, 215 strains of Candida spp (84.3%), 28 strains of Cryptococcus neoformans (11.0%), 4 strains of Talaromyces marneffei (1.6%) and 8 strains of others (3.1%). Among the Candida spp 90 strains of Candida albicans, 39 strains of Candida parapsilosis complex, 36 strains of Candida glabrata, 33 strains of Candida tropicalis, 8 strains of Candida guilliermondii, and 9 strains of other Candida. In the department, the ICU was predominant, accounting for 35.7%. The top four Candida (Candida albicans, Candida parapsilosis complex, Candida glabrata, Candida tropicalis) were analyzed for drug sensitivity, Candida albicans and Candida parapsilosis complex group were more sensitive to antifungal drugs, the sensitivity rates of Candida albicans to fluconazole, voriconazole, anidulafungin, caspofungin, micarafungin were 89.2%, 92.8%, 97.6%, 97.6%, 96.4%, respectively. The sensitivity rates of Candida parapsilosis to fluconazole and voriconazole were 89.7% and 94.9%, and to anidulafungin, caspofungin and micafungin were all 100%. Echinocandins had stronger antibacterial activity against Candida spp., Candida parapsilosis complex and Candida tropicalis had 100% sensitivity to echinocandins, Candida albicans had more than 95% sensitivity to echinocandins, and Candida glabrata had about 90% sensitivity to echinocandins. Candida tropicalis was less sensitive to fluconazole and voriconazole with 66.7% and 54.5%, and the sensitivity of Candida glabrata to fluconazole was mainly concentrated in susceptible dose dependent (SDD), accounting for 91.4%. The four Candida species did not show resistance to amphotericin B, all of them showed wild-type strains, Candida tropicalis showed the highest non-wild-type rate to posaconazole and itraconazole with 21.2% and 36.4%, and the drug sensitivity results of Cryptococcus neoformans showed that 4 out of 23 strains showed resistance to amphotericin B (non-wild-type) and 3 strains showed resistance to fluconazole (non-wild-type). Conclusions　The fungus of bloodstream infection is mainly Candida spp.. Among of them, Candida albicans accounts for the highest percentage, echinocandins have good antibacterial effect on Candida, Candida is sensitive to amphotericin B as wild type, but Candida tropicalis has slightly higher resistance rate to fluconazole and voriconazole, and the non-wild type rate of Cryptococcus neoformans to amphotericin B is increasing, and clinicians should pay high attention to the rational use of antifungal drugs.

AIMTo observe expression regularity of SMalpha-actin, SM22alpha, myocardin and SMMHC during early embryonic vascular development, and to initially investigate the differentiation effect of platelet derived growth factor-BB (PDGF-BB) on vascular smooth muscle cells (VSMCs) during that period.

METHODSMurine embryonic stem cell line expressing the enhanced green fluorescent protein (GFP) under the transcriptional control of the smooth-muscle-specific SM22alpha promoter was used to make embryoid bodies,and to analyze the expression regularity of SMalpha-actin, SM22alpha, myocardin and SMMHC by immunofluorescence stainings, RT-PCR and Western blot. Then AG1296 (PDGF receptor inhibitor) 0 micro-mol/L(control group), 10 micromol/L and 50 micromol/L were used to treat EBs respectively in order to analyze the differences of SMa-actin, SM22alpha, myocardin and SMMHC at gene and protein levels among the three groups.

RESULTSSMalpha-actin, myocardin, SM22alpha and SMMHC expression in EBs were found to begin at day 0 (ESCs), 8, 11, 13 respectively during early embryonic vascular development. There were no clear differences in SMa-actin, SM22alpha, myocardin and SMMHC protein expression and SM22alpha, myocardin and SMMHC mRNA level among the three groups of different concentrations of AG1296.

CONCLUSIONA spontaneous VSMCs differentiation occurs during EBs development, SMalpha-actin is the first to be detected,the following are myocardin, SM22a and SMMHC. PDGF-BB may not be indispensable for the regulation of expression of VSMCs markers during early EBs differentiation.

Actins ; genetics ; metabolism ; Animals ; Biomarkers ; metabolism ; Cell Differentiation ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; metabolism ; Mice ; Microfilament Proteins ; genetics ; metabolism ; Muscle Proteins ; genetics ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Nuclear Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-sis ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Trans-Activators ; genetics ; metabolism

AIMTo investigate the role of MAPK in the migration of human coronary artery smooth muscle cells(HCASMC ) into three-dimensional fibrin gels.

METHODSHCASMC were primarily cultured. HCASMC migration was measured with a phase-contrast microscope in the presence or absence of PD98059, SB203580, and SP600125, the inhibitors of ERK, p38, and JNK, respectively. Phosphorylation of ERK, p38 and JNK were analyzed by Western blotting in the presence or absence of PD98059, SB203580 or SP600125.

RESULTSHCASMC that migrated into the three-dimensional fibrin gel exhibited a characteristic elongated spindle-shaped appearance and formed vessel-like structure. The number of migrated HCASMC increased with incubation time and concentration of fibrinogen in the range between 0.8 g/L and 6.4 g/L. Western blot showed that fibrin induced phosphorylation of ERK, p38 and JNK time dependently and PD98059, SB203580 and SP600125 could inhibit their activation, respectively. Migration of HCASMC into the fibrin gels was inhibited by SP600125 20 micromol/L and SB203580 10 micromol/L, respectively. Furthermore, inhibition of SP600125 20 micromol/L had a more profound effect. PD98059 50 ,mol/L, however, failed to influence migration of HCASMC. Hence, migration of HCASMC into the fibrin gels is JNK- and p38-dependent, but not ERK-dependent.

CONCLUSIONFibrin gel induces HCASMC migration into itself by activation of JNK and p38, but not ERK, which may play an important role in pathogenesis of atherothrombosis and restenosis.

Anthracenes ; pharmacology ; Cell Movement ; Cells, Cultured ; Coronary Vessels ; cytology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibrin ; metabolism ; Flavonoids ; pharmacology ; Humans ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; MAP Kinase Signaling System ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Pyridines ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

AIMTo construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting TGF-beta1 for further research on the effects of TGF-beta1 on vasculogenesis and angiogenesis.

METHODSThree pairs of siRNA target sequences coding from the mRNA of TGF-beta1 gene were designed and three pairs of nucleotides were synthesized. After annealing, the double-strand DNA products were ligated into the pEN_mH1c entry vector, and in turn into the shRNA eukaryotic expression vector pDS_hpEy labled by GFP through the LR recombination reaction. After sequencing successfully, the three resulting TGF-beta1 shRNA expression vectors were transfected into the mouse fibroblast cell line (NIH/3T3), and then cell clones stably expressing TGF-beta1 shRNA were screened. Reverse Transcript-Polymerase Chain Reaction (RT-PCR) and Western blot were used to detect the mRNA and protein expression.

RESULTSRT-PCR and Western blot showed that one of the TGF-beta1 shRNA expression vectors pDS_Tc downregulated TGF-pl mRNA and protein expression markedly in NIH/3T3 cells.

CONCLUSIONShRNA eukaryotic expression vectors targeting TGF-beta1 are successfully constructed which can be used for further investigation on the mechanism through which TGF-beta1 regulates vasculogenesis and angiogenesis.

Animals ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; Neovascularization, Physiologic ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo investigate the effects of ulinastatin (Uti) and low-molecular-weight heparin (Lmwh) on coagulation function and deep vein thrombosis (DVT) in patients undergoing hip joint replacement.

METHODSFrom March to December 2010 150 ASAI-II patients with average age of 72.5 (65 - 85) years undergoing hip joint replacement were randomly divided into 3 groups (n = 50 each): normal saline (NS) control group (Group C), Uti group (Group U) and Lmwh group (Group L). Group U received intravenous infusion of ulinastatin (10 000 U/kg) at preoperative, perioperative and after operation 1, 2 and 3 d, respectively. Group C received the same volume of NS instead of Uti. Group L were injected Lmwh subcutaneously (3200 U/d) at preoperative, after operation 1, 2 and 3 d. Blood samples were taken before operation (T(0)), at the end of surgery (T(1)), 1 d (T(2)), 2 d (T(3)) and 3 d (T(4)) after operation for determination the values of R, K, α angle, MA and CI, using thromboelastography, and the DVT were also examined through color Doppler ultrasonography at 3 d after operation.

RESULTSCompared with T(0), R, K were shorter, α angle, MA and CI were larger in group C, the values at T(2) were up to the peak then declined at T(4). Compared with group C, the value of R, K were larger, the value of α angle, MA and CI were shorter in group U and group L. The DVT checked by ultrasonography were found in 20 cases in group C, 1 case in group U, and zero case in group L. The differences were no statistically significant between group U and group L.

CONCLUSIONIntravenous infusion of Uti during the period of operation can correct the hypercoagulability of blood and decrease the incidence of DVT after operation.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; Blood Coagulation ; Double-Blind Method ; Female ; Glycoproteins ; therapeutic use ; Heparin, Low-Molecular-Weight ; therapeutic use ; Humans ; Male ; Postoperative Complications ; prevention & control ; Venous Thrombosis ; etiology ; prevention & control

To investigate the effects and molecular mechanisms of the cellular repressor of E1A-stimulated genes (CREG) on the apoptosis of vascular smooth muscle cells (VSMCs), the human internal thoracic artery-Shenyang (HITASY) cells were infected with sense-CREG [pLNCX(2)(+)/CREG] and antisense-CREG [pLXSN(-)/CREG] retrovirus respectively. The stably infected cells were obtained by screening the G418-resistant clones. DAPI nuclei staining and Annexin V/PI FASC assay indicated that over-expression of CREG in HITASY cells infected with pLNCX(2) (+)/CREG inhibited VSMC apoptosis induced by serum deprivation, accompanied with decreased expression of caspase-9 mRNA detected by RT-PCR. Furthermore, Western blot analysis showed that p38 mitogen activated protein kinase (p38 MAPK) expression and activation were significantly enhanced in HITASY cells infected with pLNCX(2) (+)/CREG. The inhibition of CREG protein expression in cells infected with pLXSN(-)/CREG promoted the VSMC spontaneous apoptosis, as well as down-regulated p38 MAPK expression and activation, when cells were cultured with 10% fetal bovine serum (FBS) mediums. These results implicate that the CREG protein has the ability to regulate VSMC apoptosis in which the activation of p38 MAPK is possibly involved. To further identify the role of p38 MAPK in VSMC apoptosis, SB203580, a specific inhibitor of p38 MAPK, was used to inhibit p38 MAPK activity. When p38 MAPK signaling pathway was blocked, the effects that over-expression of CREG protein inhibited VSMC apoptosis disappeared. Taken together, the present work indicates that over-expression of CREG protein inhibits VSMC apoptosis, and this inhibitory effect is partly mediated by p38 MAPK signaling pathway.
Apoptosis ; physiology ; Caspase 9 ; metabolism ; Cells, Cultured ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Repressor Proteins ; genetics ; physiology ; Signal Transduction ; physiology ; p38 Mitogen-Activated Protein Kinases ; metabolism

This study was purposed to explore the expressions of platelet-activated markers PAC-1 and CD62p in peripheral blood of malignant lymphoma patients and the influence of dipyridamole on their expression. 32 lymphoma patients were divided into simple chemotherapy group (simple group) and chemotherapy plus dipyridamole group (combined group) randomly, and 15 healthy peoples were selected as control group. The dipyridamole of 100 mg/day was given to the patients in combined group. The expression levels of PAC-1, CD62p and fibrinogen (Fib) were detected by flow cytometry and magnetic bead method on day 0, 3, 7 and 14 of chemotherapy respectively. The results showed that the levels of PAC-1, CD62p and Fib in lymphoma patients were significantly higher than those in control group (p < 0.01, 0.05), moreover there was positive correlation between levels of PAC-1 and Fib (r = 0.549, p < 0.01). PAC-1 expression on day 0 and 3 of chemotherapy in simple group was higher than that on day 14 (p < 0.05, 0.01) and CD62p expression on day 3 of chemotherapy was higher than that on day 0, 7 and 14 (p < 0.05, 0.01). PAC-1 expression in combined group on day 14 of chemotherapy was lower than than on day 0 and 3 (p < 0.05, 0.01), and CD62p on day 14 was lower than that on day 3 of chemotherapy (p < 0.05); PAC-1 and CD62p expressions in combined group on day 3, 7 and 14 of chemotherapy were decreased than those in simple group, but Fib level was not changed significantly. It is concluded that the patients with malignant lymphoma usually accompany with platelet activation and hyperfibrinogenemia in peripheral blood. Applying dipyridamole routine dosage in chemotherapy can efficiently restrain platelet activation.
Adolescent ; Adult ; Aged ; Dipyridamole ; therapeutic use ; Dual Specificity Phosphatase 2 ; metabolism ; Female ; Fibrinogen ; metabolism ; Humans ; Lymphoma ; blood ; drug therapy ; Male ; Middle Aged ; P-Selectin ; metabolism ; Platelet Activation ; Young Adult

OBJECTIVETo evaluate the effect of ulinastatin on post-operative Cognition disorders in elderly patients undergoing hip joint replacement.

METHODSForty ASA I or II elderly patients undergoing selective hip joint replacement, aged > or = 65 years, were randomly divided into 2 groups (n = 40 each): control group and ulinastatin group. Ulinastatin group received iv infusion of ulinastatin (10,000 u/kg) after skin incision, (5,000 U/kg) after operation 1, 2, 3 d respectively, included 21 males and 19 females with an average age of (75.00 +/- 7.81) years old. Control group received the same volume of normal saline instead of ulinastatin, included 20 males and 20 females with an average age of (72.80 +/- 7.25) years old. Neuroeognitive testing was performed on the preoperative day and on the 3th postoperative day and post-operative cognition disorders was defined as 1 SD decline from baseline on neurocognitive assessment. Serum S100beta protein were measured before operation, at the end of surgery, 3, 24 h and 3 d after operation.

RESULTSThe incidence rate of postoperative cognition disorders was 2.5% in ulinastatin group, there were lower than those of patients in the control group (25%) (P < 0.05); In control group, the scales for MMSE before and after operation were (25.2 +/- 2.1), (22.6 +/-2.5) scores and the level of serum S100beta protein at T0-4 were (0.041 +/- 0.012), (0.125 +/- 0.031), (0.178 +/- 0.036), (0.142 +/- 0.038), (0.048 +/- 0.015) microg/L. As well in ulinastatin group, above date were (25.9 +/- 2.4), (24.8 +/- 2.1), (0.040 +/- 0.013), (0.095 +/- 0.021), (0.116 +/- 0.017), (0.087 +/- 0.019) and (0.043 +/- 0.012) respectively. Compared with preoperative, MMSE evaluation scale was decreased on the 3th postoperative day and the S100beta was increased markedly at T1-3 in control group (P < 0.05); Compared with control group, MMSE evaluation scale was increased and the S100beta was decreased markedly at T1-3 in ulinastatin group (P < 0.05 ).

CONCLUSIONIntravenous infusion of ulinastatin during operation can prevent the occurrence of POCD in elderly patients.

Aged ; Arthroplasty, Replacement, Hip ; adverse effects ; Cognition Disorders ; blood ; drug therapy ; Female ; Glycoproteins ; therapeutic use ; Humans ; Male ; Nerve Growth Factors ; blood ; Postoperative Complications ; blood ; drug therapy ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; blood

The study was aimed to investigate the clinical significance of coagulation function changes in lymphoma patients and to analyze the relationship between their changes and international prognostic index (IPI). The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) were detected by magnetic bead method in 75 lymphoma patients and 20 healthy persons. The dehydrogenase (LDH) level was detected by rate method in all lymphoma patients and healthy persons. The results showed that (1) the APTT and FIB more obviously increased in lymphoma patients which displayed as hyperfibrinogenemia, as compared with control group (p < 0.05, p < 0.01); no obvious changes of coagulation indexes presented in patients with different ages and extranodal lesions (p > 0.05, p < 0.01). (2) APTT and FIB levels in stage III and IV patients were much higher than those in the stage II (p < 0.05 and < 0.01), and FIB level in stage IV group was significantly higher than those in the stage III (p < 0.05). FIB level in symptomatic group was significantly higher than that in asymptomatic group (p < 0.01). (3) APTT and FIB in increased LDH group were obviously higher than those in control group (p < 0.05, p < 0.01). Furthermore, FIB in increased LDH group was higher than that in normal LDH group (p < 0.05). FIB in performance status (PS) 2 - 4 groups increased significantly as compared with those in PS 0-1 group (p < 0.01). (4)FIB levels in the low-middle-risk, high-middle-risk and high-risk groups were significantly higher than those in control group (p < 0.01), while FIB levels in high-middle-risk and high-risk groups were higher than those in low-risk group (p < 0.05). (5) the number of FIB increased patients in symptomatic group, increased LDH group, PS 2 - 4 group and Ann Arbor stage III-IV group were much higher than those in counterparts (p < 0.05 or 0.01).There were positive correlations between FIB and LDH level, PS grades, Ann Arbor stages as well as risk grades respectively (p < 0.05 or 0.01). It is concluded that lymphoma patients usually accompany with hyperfibrinogenemia which may be influenced by Ann Arbor stage, systemic symptom, LDH level and PS grade. FIB is supposed to be an effective indication of prognosis in lymphoma patients.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Coagulation ; Case-Control Studies ; Female ; Humans ; Lymphoma ; diagnosis ; pathology ; physiopathology ; Male ; Middle Aged ; Neoplasm Staging ; Partial Thromboplastin Time ; Prognosis ; Prothrombin Time ; Young Adult

OBJECTIVETo investigate the effects of caffeic acid ester fraction (Caf) from Erigeron breviscapus, mainly composed of dicaffeoylquinic acids (diCQAs), on microglial activation in vitro and focal cerebral ischemia in vivo.

METHODSThe production of nitric oxide (NO), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) induced by lipopolysaccharide (LPS) treatment in rat primary cultured microglia were measured by Griess reaction or enzyme-linked immunosorbent assay. Cell viability of cortical neurons was measured using AlamarBlue reagent. The behavioral tests and the infarct area of brain were used to evaluate the damage to central nervous system in rat middle cerebral artery occlusion (MCAO) model of cerebral ischemia. Real time polymerase chain reaction was used to determine the expression of inducible nitric oxide synthase (iNOS), TNF-α and IL-1β mRNA in ischemic cerebral tissues.

RESULTSCaf inhibited the production of NO, TNF-α and IL-1β induced by LPS treatment in primary microglia in a dose-dependent manner. Exposure of cortical neurons to conditioned medium from Caf-treated microglia increased neuronal cell viability (P<0.01) compared with conditioned medium from LPS-treated alone. In MCAO rat model of cerebral ischemia, Caf could significantly improve neurobehavioural performance and reduce percentage infarct volume compared with the vehicle group (P<0.05). Caf could also significantly inhibit the up-regulation of iNOS, TNF-α, and IL-1β gene expressions in ischemic cerebral tissues.

CONCLUSIONCaf could suppress microglial activation, which may be one mechanism of its neuroprotective effect against ischemia.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Brain Ischemia ; complications ; drug therapy ; pathology ; prevention & control ; Caffeic Acids ; chemistry ; pharmacology ; Chemical Fractionation ; Chromatography, High Pressure Liquid ; Erigeron ; chemistry ; Gene Expression Regulation ; drug effects ; Infarction, Middle Cerebral Artery ; complications ; pathology ; Interleukin-1beta ; genetics ; metabolism ; Microglia ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; chemistry ; pharmacology ; therapeutic use ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Plant Extracts ; pharmacology ; Quinic Acid ; analogs & derivatives ; chemistry ; pharmacology ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; metabolism