Objective Two hundred and eighty seven strains of fungi isolated from the cutaneous lesions in patients with chromoblastomycosis from Zhang Qiu county and its surrounding region were observed and analysed. Methods The fungi were cultured and the morphologic characters of colonies were observed under scanning electron microscope. Results There were 283 strains of C.carrionii, 2 strains of F.pedrosoi, 1 strain of P.verrucosa and 1 strain of undeterminate phialophora sp.were identificated. Conclusion The predominant pathogenic fungi isolated from patients with chromoblastomycosis in Zhang Qiu county and its surrounding region were C.carrionii which confirms that the pathogenic fungi in nothern China is C.carrionii.

To investigate the chemical constituents of the endophytic fungus Penicillium sp. FJ-1 of Ceriops tagal, the chemical constituents were isolated by column chromatography on silica gel and Sephadex LH-20. Their structures were elucidated on the basis of spectroscopic analysis. Their antibacterial activity was tested by paper disco diffusion method. Two compounds were isolated and identified as 7-hydroxy-deoxytalaroflavone (1), and deoxytalaroflavone (2). Compound 1 is a new compound, and compounds 1 and 2 showed weak activity against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.

Objective To compare the carriage of dematiaceous fungi in wild toads from both en-demic area and non-epidemic area of chromoblastomycosis.Methods A total of354and45wild toads(Bufo Chinese)were caught from two counties of Shandong Province,Zhangqiu county,an endemic area of chromoblastomycosis,and from Laiyang county,an area where the infection of dematiaceous fungi in humans has not been found,respectively.The toads were killed and the suspension of the internal organs were used to isolate dematiaceous fungi.Results Eleven species including162strains of dematiaceous fungi were iso-lated from151of354(42.7%)toads from the endemic area.The isolated strains consisted of49Phialopho-ra verrucosa,33Exsophiala,19Phialophora(nonpathogenic),19E.jenselmei,12Cladosporium carrionii,3suspected of C.carrionii,7Rhinocladiella sp.,5Fonsecaea pedrosoi,4C.trichoides,3E.spinifera,1C.mi-nourae,4unidentified.Only4nonpathogenic dematiaceous fungi were isolated from4of45(8.9%)toads obtained from non-epidemic area.Conclusions Varieties of pathogenic dematiaceous fungi,especially C.carrionii,are isolated from wild toads in endemic area of chromoblastomycosis,while none from non-endemic area.The results suggest that the colonization of pathogenic dematiaceous fungi in wild toads may be associa-ted with endemicity of chromoblastomycosis.

Abstract BACKGROUND: Alveolar bone deficiency wil not meet aesthetic and functional requirements for dental implants. OBJECTIVE:To observe the repair effect of passage 3 autologous bone marrow mesenchymal stem cels (BMSCs) and platelet-rich fibrin (PRF) on alveolar bone defects in rabbits. METHODS:Twenty-seven New Zealand rabbits were randomly divided into BMSCs/PRF group, PRF group and model group (n=9 per group). The left mandible incisors were extracted in al the rabbits under general anesthesia. BMSCs/PRF group was immediately implanted BMSCs/PRF composite into the alveolar socket, PRF group only implanted PRF, and model group implanted nothing. RESULTS AND CONCLUSION: In the model group, the alveolar crest and alveolar mucosa become sunken notably and narrowed. In the BMSCs/PRF and PRF groups, the thickness of alveolar bone wal, alveolar bone width, alveolar bone height difference, and bone mineral density were al increased, especialy in the former group. In addition, the trabecular arrangement was better in the BMSCs/PRF groups than the model and PRF group. Our findings indicate that alveolar socket filing with composite of BMSCs and PRF can achieve preservation of alveolar bone width and height after tooth extraction in rabbits.

Objective To construct human lung cancer cell model with human MutS homologous protein 2 (hMSH2) overexpression for exploring the effect of hMSH2 molecule in the cytotoxicity of γδ T cell against lung cancer cells.Methods hMSH2 coding sequence was cloned by PCR for construction of recombinant vector which over expressed hMSH2-EGFP fusion protein using homologous recombination.The recombinant vector was transfected to lung cancer cell line NCI-H520 to construct human lung cancer cell model overexpressing hMSH2 molecule.The expression of hMSH2 molecule in NCI-H520 was detected by Western blot.The expression of hMSH2 on the cell membrane was measured by flow cytometry.Cytotoxicity of expanded γδ T cells from peripheral blood mononuclear cells against NCI-H520 cells was detected by LDH release assay in vitro.Results hMSH2 coding sequence (2805 bp) was cloned and the result of restriction endonuclease digestion of Fugw-hMSH2 recombinant vector was accordance with the anticipated objective strip size.Exogenous hMSH2-EGFP fusion protein was expressed in NCIH520 cells.The level of hMSH2 molecule on the surface of NCI-H520 cells with overexpression of hMSH2 was significantly increased (P＜0.001).Cytotoxicity of γδ T cells against NCI-H520 cells with overexpression of hMSH2 was significantly increased compared to the wild type NCI-H520 cells (P＜0.05).Conclusions Lung cancer cell model that overexpresses hMSH2 molecule is successfully constructed,hMSH2 molecule on the cell membrane is increased and the cytotoxicity of γδ T cells against lung cancer cells is enhanced.

Objective To study the role of human MutS homolog 2 (hMSH2), a newly identified protein ligand that was recognized by Vγ9δ2 T cells , in innate anti-gastric carcinoma immunity .Methods Flow cytometry and laser confocal microscopy were used to identify hMSH 2 that ectopically expressed on gas-tric carcinoma cell line 803.An anti-hMSH2 antibody was used to block hMSH2 to evaluate its effects on the cytotoxicity of Vγ9δ2 T cells and their cytokines secretion .Subcellular distribution of hMSH 2 in gastric car-cinoma tissues was examined by tissue microarray immunohistochemistry analysis .Results Ectopic mem-brane expression of hMSH 2 was observed on 803 cells at a relatively high level .Vγ9δ2 T cells blocked with specific anti-hMSH2 antibody showed a decreased cytotoxicity and a reduced IFN-γbut an increased TNF-αsecretion.Ectopic expression of hMSH2 was found in various types of gastric carcinoma tissues at different stages.Enhanced expression of hMSH2 was detected in specimens collected from patients with chronic super-ficial gastritis.Conclusion Ectopically expressed hMSH2 served as a stress-induced endogenous ligand which could promote the cytotoxicity of Vγ9δ2 T cells against gastric carcinoma cells and enhance their IFN-γsecretion.hMSH2 played an essential role in innate anti-gastric carcinoma immunity .

Objective To investigate the expression and im plication of cell-cycle related factors(Cyclin D1,CDK4,P16 MTS1 and PRb)in condyloma acuminatum(CA).Methods The expression and distribution of cyclin D1,CDK4,P16 MTS1 and PRb were detected by immunohisto chemical technique streptavidin peroxidase(SP)in 27cases of HPV6/11positive CA that were confirmed by PCR and 10cases of norm al skins(foreskins).Results①Cyclin D1was not expressed in both normal skin and lesions of CA.②The intensity of expression of CDK4was 2.19in the basal cells in the lesions of CA,which was significantly weaker than that in normal skin(4.8,P

Objective:To analyze the survival and prognostic factors of adult acute lymphoblastic leukemia (ALL) with different consolidation regimens after complete remission by induction therapy.Methods:A total of 93 adult patients with ALL were enrolled from January 2012 to June 2019 in Peking University Shenzhen Hospital. All the patients achieved complete remission induced by VDLCP regimen, and were divided into the standard group, intensive group and transplantation group according to the consolidation treatment. Thirty-four patients in the standard group received an ALL-like chemotherapy regimen based on VDLCP or Hyper-CVAD consolidation for 4-6 courses. Twenty-nine patients in the intensive group received BFM90/95 consolidation treatment for 2 years. Thirty patients in the transplantation group received allogeneic hematopoietic stem cell transplantation (allo-HSCT) after 2-3 courses of consolidation with the original induction regimen. The median follow-up was 18 (3-96) months, and the main follow-up indicators were overall survival (OS) and disease free survival (DFS). Prognostic factors of adult ALL patients and treatment-related deaths in each group were analyzed.Results:The 3-year OS rates of the standard group, intensive group and transplantation group were 54.0% (95% CI: 35.3%-72.6%), 71.8% (95% CI: 41.0%-94.5%), 62.3% (95% CI: 43.6%-80.9%), with a statistically significant difference ( χ2=6.110, P=0.047). The 3-year DFS rates of the three groups were 31.4% (95% CI: 12.9%-49.8%), 72.1% (95% CI: 52.3%-91.9%), 65.7% (95% CI: 45.3%-86.1%), with a statistically significant difference ( χ2=13.831, P=0.001). There were no significant differences in OS and DFS between the intensive group and the transplantation group ( χ2=0.709, P=0.400; χ2=0.046, P=0.830). OS and DFS of the intensive group were better than those of the standard group ( χ2=5.346, P=0.021; χ2=10.326, P=0.010). Multivariate analysis suggested that bone marrow minimal residual disease (MRD) negative on day 14-21 of chemotherapy was an independent prognostic factor affecting adult ALL ( HR=0.114, 95% CI: 0.015-0.841, P=0.033). The 3-year OS rates of Ph + ALL patients who received and did not receive allo-HSCT were 53.5% (95% CI: 23.1%-83.8%), 52.4% (95% CI: 23.8%-81.0%), the 3-year DFS rates were 77.1% (95% CI: 54.2%-100.0%), 35.0% (95% CI: 4.8%-65.2%), and there were no significant differences between the two groups ( χ2=3.600, P=0.223; χ2=3.824, P=0.050). The treatment-related mortalities of the non-transplantation group (standard group + intensive group) and the transplantation group were 3.2% (2/63) and 20.0% (6/30), and the treatment-related mortality of the non-transplantation group was significantly lower than that of the transplantation group ( χ2=7.318, P=0.007). Conclusion:Adult ALL has a poor prognosis. The 3-year OS rate and 3-year DFS rate of BFM intensive consolidation therapy are better than those of standard consolidation therapy, achieving a similar effect to allo-HSCT, but treatment-related mortality does not increase significantly. Patients with bone marrow MRD negative on the day 14-21 of chemotherapy have the better OS and DFS.

Objective To evaluate the effect of resuscitation with Ringer′s malate solution on acute lung injury caused by hemorrhagic shock in rats. Methods Forty-eight SPF healthy male Sprague-Dawley rats, aged 7-9 weeks, weighing 280-320 g, were assigned into 4 groups ( n＝12 each) using a random number table method: sham operation group (group S), normal saline group (group NS), Ringer′s lac-tate solution group ( group RL) and Ringer′s malate solution group ( group RM). In NS, RL and RM groups, the model of hemorrhagic shock was established, and rats were resuscitated after 60 min of hemor-rhagic shock. Rats were sacrificed at 3 h after resuscitation, and bronchial alveolar lavage fluid ( BALF) was collected to count neutrophils. Lung tissues were obtained for determination of the wet∕dry weight ratio (W∕D ratio), myeloperoxidase (MPO) activity, and contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6. Lung tissues were obtained for examination of the pathological changes. Results Compared with group S, the neutrophil count in BALF, W∕D ratio, MPO activity and contents of TNF-α, IL-1β and IL-6 were significantly increased in NS, RL and RM groups ( P＜0. 05). Compared with NS and RL groups, the neutrophil count in BALF, W∕D ratio, MPO activity and contents of TNF-α, IL-1β and IL-6 were significantly decreased in NS and RL groups (P＜0. 05). Conclusion The severity of acute lung injury is reduced when Ringer′s malate solution is used for resuscitation as compared with that when normal saline and Ringer′s lactate solution are used in a rat model of hemorrhagic shock.

Objective: To investigate the incidence of microvascular myocardial ischemia in diabetic patients without obstructive coronary artery disease (CAD) and its relationship with angina. Materials and Methods: Diabetic patients and an intermediate-to-high pretest probability of CAD were prospectively enrolled. Non-diabetic patients but with an intermediate-to-high pretest probability of CAD were retrospectively included as controls. The patients underwent dynamic computed tomography-myocardial perfusion imaging (CT-MPI) and coronary computed tomography angiography (CCTA) to quantify coronary stenosis, myocardial blood flow (MBF), and extracellular volume (ECV). The proportion of patients with microvascular myocardial ischemia, defined as any myocardial segment with a mean MBF ≤ of 100 mL/min/100 mL, in patients without obstructive CAD (Coronary Artery Disease–Reporting and Data System [CAD-RADS] grade 0–2 on CCTA) was determined. Various quantitative parameters of the patients with and without diabetes without obstructive CAD were compared. Multivariable analysis was used to determine the association between microvascular myocardial ischemia and angina symptoms in diabetic patients without obstructive CAD. Results: One hundred and fifty-two diabetic patients (mean age: 59.7 ± 10.7; 77 males) and 266 non-diabetic patients (62.0 ± 12.3; 167 males) were enrolled; CCTA revealed 113 and 155 patients without obstructive CAD, respectively. For patients without obstructive CAD, the mean global MBF was significantly lower for those with diabetes than for those without (152.8 mL/min/100 mL vs. 170.4 mL/min/100 mL, P < 0.001). The mean ECV was significantly higher for diabetic patients (27.2% vs. 25.8%, P = 0.009). Among the patients without obstructive CAD, the incidence of microvascular myocardial ischemia (36.3% [41/113] vs. 10.3% [16/155], P < 0.001) and interstitial fibrosis (69.9% [79/113] vs. 33.3% [8/24], P = 0.001) were significantly higher in diabetic patients than in the controls. The presence of microvascular myocardial ischemia was independently associated with angina symptoms (adjusted odds ratio = 3.439, P = 0.037) in diabetic patients but without obstructive CAD. Conclusion Dynamic CT-MPI + CCTA revealed a high incidence of microvascular myocardial ischemia in diabetic patients without obstructive CAD. Microvascular myocardial ischemia is strongly associated with angina.