Objective To analyze the antibiotic resistance of Branhemella catarrhalis strains isolated from sputum specimens of patients with lower respiratory tract infections from Linyi, Shandong Province, and to explore the relationship between bro genotypes of the strains and their resistance to antibiotic agents.Methods Sputum specimens were colleted from the patients with lower respiratory tract infections in Linyi People ’ s Hospital from the January 2010 to December 2014.The specimens were inoculated into 4 different disks for bacterial isolation and cultivation.β-lactamase detection and drug sensitivity tests were performed, and PCR coupled with restriction endonuclease analysis was employed for bro genotyping.χ2 test was used to compare drug resistance of strains with different bro genotypes.Results A total of 497 Branhemella catarrhalis strains were isolated in five years, among which 221 strains were isolated in winter.All strains were sensitive to ertapenem and chloramphenicol, and the resistance rates to amoxicillin/clavulanate and cefaclor were low (≤2.8%).The strains were highly resistant to compound sulfamethoxazole, erythromycin and ampicillin (47.6%-89.8%), and there was a trend of increasing resistance rates with the year, but no statistically significant difference was observed ( P >0.05 ) .β-lactamases was positive in 412 strains (82.9%), and all of these strains were positive for bro gene, and the resistances to erythromycin, compound sulfamethoxazole, levofloxacin and ampicillin were higher in bro positive strains than those in bro negative strains (χ2 =12.16, 16.18, 8.41 and 200.00,P<0.05).Among bro positive strains, 391 (94.9%) were of genotype bro-1, 21 (5.1%) were of genotype bro-2, and their resistance to antibiotic agents was not of statistical difference ( P >0.05 ).Conclusions Most of Branhemella catarrhalis clinical isolates are β-lactamase producing strains, and bro-1 is the most common genotype.Strains are highly sensitive to carbapenems, cephalosporins andβ-Lactamaseinhibitors, which can be recommended for the treatment of Branhemella catarrhalis-related respiratory tract infections.

Objective: To investigate the expression of ATP-Binding Cassette Proteins including P-gp (P-glycoprotein), MRP1 (multidrug resistance associated protein 1) and BCRP (breast cancer resistance protein) in hepatocellular carcinoma and its relationship with pathological features. Methods: The expression of P-gp/MDR1 (multidrug resistance gene 1), MRP1 and BCRP in hepatocellular carcinoma was examined by RT-PCR and immunohistochemistry in 34 cases of hepatocellular carcinoma and 19 cases of paraneoplastic hepatic tissues. Results: The expression of MDR1, MRP1 and BCRP mRNA (messenger ribonucleic acid) was 1.15±0.24, 0.64±0.33, and 1.07±0.32 in hepatocellular carcinoma and 0.36±0.14, 0.19±0.06, and 0.31±0.09 in paraneoplastic hepatic tissues. The expression of MDR1, MRP1 and BCRP mRNA was 1.38±0.26, 0.73±0.35, and 1.34±0.21 in poorly differentiated hepatocellular carcinoma and 0.74±0.32, 0.30±0.11, and 0.45±0.13 in well differentiated hepatic tissues. The immunohistochemical positive substance was detected in the plasma membrane and cytoplasm. The positive rates of P-gp, MRP1 and BCRP were 82.35%, 58.82%, and 79.41% in hepatocellular carcinoma and 42.11%, 26.32%, and 36.84% in paraneoplastic hepatic tissues, respectively. The positive rates of P-gp, MRP1 and BCRP were 100.00%, 81.25%, and 100.00% in poorly differentiated hepatocellular carcinoma and 66.67%, 38.89%, and 61.11% in well differentiated hepatic tissues. The expression of three indicies in hepatocellular carcinoma was higher than that in paraneoplastic hepatic tissues (P<0.05). The expression of P-gp/MDR1, MRP1 and BCRP in poorly differentiated hepatocellular carcinoma was higher than that in well differentiated hepatic tissues (P<0.05). No correlation was found among the three indices. Conclusion: Intrinsic multidrug resistance exsists in hepatocellular carcinoma, with various mechanisms. The multidrug resistance of HCC (hepatic cell carcinoma) is related to P-gp/MDR1, MRP1 and BCRP. MRP1 and BCRP may be targets for reversing multidrug resistance.

BACKGROUND: Studies of biological characteristics of mesenchymal stem cells (MSCs) and regulatory factors that influenced the differentiation of MSCs have shown that the proportion of the natural differentiation from in vitro primarily cultured MSCs into hepatocytes was low, and to select a suitable inductor is important to enhance the differentiation of MSCs into hepatocytes.OBJECTIVE: To verify the feasibility of induced differentiation of rat bone marrow MSCs (BMSCs) into hepatocytes using the combination of hepatocyte growth factor (HGF), epidermal growth factor (EGF) and fibroblast growth factor (FGF-4).DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Experimental Center, Weifang Medical College in August 2007.MATERIALS: Totally 40 Sprague-Dawley rats were supplied by the Experimental Animal Center, Weifang Medical College.METHODS: Rat BMSCs were incubated by adherent method. BMSCs at passage 3 were assigned to 2 groups. BMSCs in the blank control group were treated with L-DMEM containing 10% fetal bovine serum. BMSCs in the combination group were treated with 10 μg/L FGF, 8 μg/L HGF and 8 μg/L EGF following above-mentioned procedures.MAIN OUTCOME MEASURES: Inverted microscope was used to observe the morphological changes in cells.Immunofluorescence method was used to observe the expression of alpha-fetoprotein (AFP) and albumin (ALB). PAS was employed to detect the expression of glycogen. Fox green intake experiment was conducted. Enzymology was utilized to test the contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP).RESULTS: BMSCs in the combination group presented polygonal, orbicular or round shape. BMSCs in the blank control group remained spindle. BMSCs in the combination group were positive for AFP and ALB at day 14 following culture, and a few PAS-positive and fox green-positive cells were found at day 7. Positive cells became more over time. Synthesis of ALT, AST and ALP was detected at day 14, reached a peak at 21 days, and then decreased. Above-described indexes were negative in the blank control group.CONCLUSION: After induced by the FGF, HGF and EGF, BMSCs have the ability to differentiate into hepatocytes in vitro.

Objective To investigate the expression of hepatocyte nuclear factor-1α (HNF-1α) and hepatocyte nuclear factor-4α (HNF-4α) in human hepatocellular carcinoma (HCC) and explore the function of HNF-1α and HNF-4α during HCC carcinogenesis and development. Methods Twenty-six specimens of hepatocellular carcinoma were collected. The expressions of HNF-1α and HNF-4α in HCC tissues and adjacent non-cancerous tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry staining. Results The mRNA levels of HNF-1α and HNF-4α were significantly lower in HCC tissues than that in adjacent non-cancerous tissues (0.818±0.371 vs. 0.383±0.102 for HNF-1α, P＜0.05;0.846±0.384 vs. 0.397±0.105 for HNF-4α, P＜0.05).The positive rates of HNF-1α and HNF-4α protein were significantly lower in HCC tissues than in adjacent non-cancerous tissues (92.3% vs. 42.3% for HNF-1α, P＜0.05;96.2% vs. 50.0% for HNF-4α, P＜0.05). The mRNA and protein expressions of HNF-1α and HNF-4α were correlated with tumor differentiation (P＜0.05). There was a negative correlation between HNF-1α and HNF-4α mRNA expressions in HCC tissues.Conclusion The expressions of HNF-1α and HNF-4α are down-regulated in HCC, which might be related to carcinogenesis and development of HCC.

Objective To explore the possibility that rat bone mesenchymal cells (BMSCs) can differentiate into hepatocytes under the affection of fetal liver filtrate. Methods PAS and green indigo dye were used to detect glycogen and differential level of hepatocytes, respectively. The concentration of ALT, AST, ALP in the culture supernatant were served as markers of hepaocyte function. Results Fourteen days after induced by the fetal liver filtrate, BMSCs changed their shapes into polygon, oval or round. Some of BMSCs were positive for AFP and ALB at 7 days after induction, then the number of positive cells increased, and most of BMSCs expressed AFP and ALB till 21days. The PAS reaction and indocyanine green(ICG) intaking also appeared at 7days. Enzyme in supernatant such as ALT, AST, ALP were fristly detected at 7days and peaked at 14days,then the level declined. Conclusion The fetal rat liver filtrate was able to induce BMSCs into cells with function and characteristics of hepatocytes.

BACKGROUND: Studies have shown that adipose-derived stem cells have pluripotent differentiation potential, but only 30%-40% of cells can differentiate into mature adipocytes with low adipogenic differentiation potential. Therefore, how to improve the adipogenic differentiation ability of adipose-derived stem cells is a key problem to be solved in the process of soft tissue regeneration. OBJECTIVE: To observe the relationship between the surface marker CD54 of rabbit adipose-derived stem cells and their adipogenic capacity, and to explore the adipogenic differentiation of CD54+/CD54- adipose-derived stem cells underthe same induction. METHODS: We successfully isolated and cultured the adipose-derived stem cells from inguinal subcutaneous fat pads (3 ml) of New Zealand white rabbits, aged 8-12 weeks, which were induced into multi-differentiation and used to detectsurface markers. We sorted the passage 3 adipose-derived stem cells by immunomagnetic beads and divided into two categories including CD54+ and CD54- adipose-derived stem cells. After 14 days of adipogenic induction, the cells in the two groups were subjected to oil red O staining and were compared by detecting the density of mature adipocytes and lipid droplet contenT.RESULTS AND CONCLUSION: The cultured adipose-derived stem cells possessed the characteristics of mesenchymal stem cells that could differentiate into mature adipocytes, osteoblasts and chondrocytes, with CD29, CD44, CD49d, CD54, CD73, CD90 and CD105 positive expression while CD31, CD34 and CD45 negative expression. Fourteen days after adipogenic induction, the density of mature adipocytes and the intracellular lipid droplet content in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). We also found that the mRNA expressions of PPARγ,ADD1, C/EBPα related to adipogenic differentiation in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). Taken together, CD54+ adipose-derived stem cells have excellent adipogenic differentiation capacity.

Objective To explore the effects of Caspase-3 and Glypicante1 (Gpc1) altered expressions on the proliferation of glioma cells and its mechanism.Methods Fifty-two glioma specimens and 13 normal cerebral tissues were collected in our hospital from October 2011 to October 2014;the protein expressions of Caspase-3 and Gpc1 in these tissues were detected with immunohistochemistry.The peDNA3.1/Gpc 1 high expression plasmids were constructed;human glioma A172 cell lines were routinely cultured in vitro;the signal path changes of Hedgehog (H-h) in the A172 cells after being transfected with 0.5,1,2 and 4 μg peDNA3.1/Gpc1 high expression plasmids were detected by Luciferase luciferase reporter gene kit.Gpc 1-1694 interference plasmids were transfected into the A172 cells to silence the Gpc1 expression and these A172 cells were given 8 iμg/mL Blasticidin (experimental group) for 24 h;normal A172 cells were given the same volume of solvent (control group);altered expressions of G-pc 1 were detected by immumohistochemical staining and proliferation activity of the cells were observed by MTT assay 24,48,72 and 96 h after transfection.Results The expressions of Caspase-3 and Gpc1 in the glioma specimens were significantly higher than those in the normal brain tissues (P＜0.05);in the glioma specimens,positive correlation between Caspase-3 and Gpc1 was noted (r=0.486,P=0.000).As compared with the control group,increased fluorescence intensity was observed in the 0.5,1,2 and 4 μg peDNA3.1/Gpc1 high expression plasmids groups (P＜0.05);immumohistochemical staining showed that CyclinD1 expression in the Hh pathway of in the peDNA 3.1/Gpc1 high expression plasmids groups was significantly lower than that in the control group;MTT assay showed that the proliferation activity of the A172 cells in the peDNA3.1/Gpc1 high expression plasmids groups at 24,48,72 and 96 h after transfection was significantly lower than that in the control group (P＜0.05).Conclusion Caspase-3 and Gpc1 play important roles in regulating the proliferation of glioma cells,which might be related to Hedgehog signaling pathway.

Objective:To ulteriorly explore the differences of psychotic symptoms and neurocognitive between patients with first-episode deficit subtype of schizophrenia (FDS) and patients with first-episode nondeficit subtype of schizophrenia (FNDS).Methods:From January 2021 to September 2021, a total of 88 first-episode treatment-naive schizophrenia were recruited from the Mental Health Center of West China Hospital and divided into FDS group( n=44) and FNDS group( n=44) according to the schedule for the deficit syndrome (SDS), and 44 healthy subjects were included as healthy control group (HC group, n=44). Positive and negative syndrome scale (PANSS) was used to assess psychotic symptoms of patients and Wechsler adult intelligence scale, trail making test and logic memory test were used to evaluate intelligence quotient and neurocognitive function of all subjects.SPSS 22.0 was used for statistical analysis, and independent samples t-test and one-way analysis of variance (ANOVA) were used to compare variables that met normal distribution, while the Mann-Whitney U test and Kruskal-Wallis H test were used to compare variables that did not meet normal distribution. Results:(1) There were significant differences in psychotic symptoms between the FDS group and the FNDS group.Compared with the FNDS group, the FDS group had higher total score of PANSS ((95.95±16.82) vs (88.39±16.29)), negative symptoms ((27.57±7.52) vs (16.57±5.76)) and anergastic reaction ((13.43±3.82) vs (7.00(5.00, 9.00)), and lower positive symptoms scores ((21.95±6.88) vs (25.41±6.07)), activation ((8.00(5.00, 9.00) vs (9.27±3.47)), depression ((5.50(4.00, 9.00) vs (8.00(6.00, 12.00)) and supplementary item ((13.60±4.17) vs (17.30±5.39))(all P<0.05). (2) There were differences in neurocognitive functions between FDS group and FNDS group, and which in FDS and FNDS group were worse than that in HC group.Spatial memory (block design test: (23.70±11.05) vs (31.72±11.49)) and information processing speed (digit symbol test: (38.38±15.85) vs (47.97±14.99)) of FDS group were significantly lower than those of FNDS group(both P<0.05). Intelligence quotient, information processing speed and spatial memory of FDS group and FNDS group were lower than those of HC group(all P<0.05). Conclusion:FDS patients has more severe negative symptoms and anergastic reaction, and exit worse information processing speed and spatial memory dysfunction than FNDS patients.This unique pattern of impairment suggests that information processing speed and spatial memory may be important classification indicators for differentiating the deficit subtype of schizophrenia in the early stage.

Objective To investigate the significance of carcinoembryonic antigen (CEA),carbohydrate antigen 19-9 (CA19-9),carbohydrate antigen 125 (CA125),neuron specific enolase (NSE),cytokeratin 19 fragment (CYFRA21-1) and squamous cell carcinoma antigen (SCC) for evaluation of first-line chemotherapeutic response and the prognostic value of these markers for prediction of overall survival (OS) in patients with advanced lung cancer.Methods Patients diagnosed with Ⅲ b/Ⅳ stage untreated,primary lung cancer and received first-line chemotherapy in Peking University Cancer Hospital & Institute from June 2013 to December 2014 were enrolled retrospectively into this study.The results of tumor markers before and after two cycles of chemotherapy and the clinical data of 181 eligible patients,including 133 males and 48 females with the average age of 58 years,were collected.The serum levels of six tumor markers were measured by electrochemiluminescence assay.Using RECISTv1.1 as standard,the sensitivity and specificity of tumor markers in classifying PR,SD,PD,were observed.The Kappa agreement test was used to evaluate the correlation between serum tumor markers and CT in evaluating chemotherapy response.The follow-up of OS was derived by telephone.Results The top three positive rates of biomarkers were CEA,CA125 and NSE in adenocarcinoma patients,CYFRA21-1,NSE and SCC in squamous carcinoma patients,NSE,CA125 and CYFRA21-1 in small cell lung cancer patients,respectively.In Kappa agreement test,the changes of serum levels of CEA and CA125,CYFRA21-1 and SCC,NSE were significantly correlated with CT for chemotherapy response evaluation in the mention above three kinds of carcinomas respectively (P ＜ 0.05).The sensitivity of tumor markers predicting PR was more than 90％,but the specificity was only 20％-58.3％,meanwhile,the specificity for SD and PD was high,but the sensitivity was low 0-66.7％).In the prognostic evaluation based on RECISTv1.1,there was no statistical significance for classifying PR,SD and PD in adenocarcinoma patients (median overall survival 17 months vs 22 months vs 14 months,P ＞0.05),but the OS of PR was longer than SD and PD in squamous and small cell lung carcinoma (median overall survival 28 months vs 22 months vs 4 months,12 months vs 8 months vs 8 months,P ＜ 0.05).Of these six tumor markers,only SCC was of statistical significance for classifying PR,SD and PD in squamous carcinoma (median overall survival 30 months vs 1 1 months vs 4 months,P ＜ 0.05).Conclusions Tumor marker has high sensitivity for predicting PR,but the value of tumor marker predicting SD and PD of advanced lung cancer was limited.The evaluation of patients would be comprehensive by combined use of tumor markers and CT.The changes of SCC after 2 cycles of chemotherapy are predictive of survival in squamous cell carcinoma patients.

Objective To explore the diagnostic value of serum neuronal Per-Arnt-Sim domain protein 4(NPASDP-4)and myelin basic protein(MBP)expression in patients with Parkinson's disease in relation to cognitive impairment(CI)and severity.Methods Selected and 138 Parkinson's disease patients admitted to the 909th Hospital of the Joint Logistics Support Force of the People's Liberation Army of China as the Parkinson's disease group,and 69 healthy people in the physical examination center of the hospital were in the healthy control group.Patients with Parkinson's disease were divided into normal cognitive function group(n=55),mild CI group(n=51)and dementia group(n=32)according to whether CI occurred and its severity.General data of subjects was collected,the serum levels of NPASDP-4 and MBP were detected by ELISA,correlation analysis was adopted by Spearman rank correlation or Pearson linear correlation,diagnostic value was analyzed by ROC curve,and influencing factors were analyzed by multivariate Logistic regression.Results Compared with the healthy control group,the levels of serum NPASDP-4(6.75±0.48 ng/ml vs 2.38±0.31 ng/ml)and MBP(8.34±0.65 μg/L vs 3.54±0.42 μg/L)in the Parkinson's disease group were increased with statistical significance(r=68.751,55.761,all P＜0.05).There were significant differences in H-Y stage among the normal cognitive function group,mild CI group and dementia group(x2=7.788,P＜0.05).Compared with the group with normal cognitive function(47.92±11.63 score),the mild CI group(50.78±13.69 score)and the dementia group(41.95±10.36 score)showed an increase in UPDRS-Ⅲ scores,and the differences were statistically significant(H=6.672,all P＜0.05).In normal cognitive function group,mild CI group and dementia group,the course of disease,and serum NPASDP-4(5.89±0.40,6.83±0.55,8.12±0.54 ng/ml)and MBP(6.65±0.56,8.94±0.69,10.27±0.70μg/L)levels were significantly increased(H=207.950,355.594,allP＜0.05),while MMSE score(28.47±0.94,24.51±1.35,17.09±2.57 score),MoCA score(27.45±1.03,20.18±1.92,11.75±2.53 score)and GPCOG total score(13.47±0.69,10.25±1.04,8.97±0.82 score)were significantly decreased,and the differences were statistically significant(H=515.005,775.933,327.584,all P＜0.05),respectively.The serum levels of NPASDP-4 and MBP in Parkinson's disease patients were significantly positively correlated with the course of disease(r=0.316,0.358),H-Y stage(r=0.345,0.384)and UPDRS-Ⅲ score(r=0.371,0.396),and significantly negatively correlated with MMSE score(r=-0.468,-0.517),MoCA score(r=-0.504,-0.569)and GPCOG total score(r=-0.527,-0.538)(all P＜0.05),respectivey.The areas under the curve(AUC)of the serum levels of NPASDP-4,MBP and their combination in diagnosing of Parkinson's disease were 0.850,0.930 and 0.960,respectively.The AUC of the serum levels of NPASDP-4 and MBP and their combination in diagnosing the severity of CI in patients with Parkinson's disease were 0.866,0.803 and 0.933,respectively.H-Y stage metaphase[OR(95％CI):4.725(1.742～12.814)],H-Y stage advanced[OR(95％CI):5.083(1.919～13.464)],UPDRS-Ⅲ score[OR(95％CI):3.257(1.464～7.246)],NPASDP-4[OR(95％CI):5.324(1.516～18.701)]and MBP[OR(95％CI):5.769(2.459～13.533)]were the influential factors for CI in patients with Parkinson's disease(all P＜0.05).NPASDP-4[OR(95％CI):4.768(2.382～9.543)]and MBP[OR(95％CI);5.846(3.141～10.882)]were the influential factors for the severity of CI in patients with Parkinson's disease(all P＜0.05).Conclusion The serum levels of NPASDP-4 and MBP in patients with Parkinson's disease were high,and they were closely related to CI and its severity,which may have certain clinical diagnostic value.