Objective To explore the clinical significance of immunocyte subsets before and after immunosuppressive therapy in the peripheral blood of patients with immune thrombocytopenic purpura (ITP).MethodsThe percentages of immunocyte subsets in the peripheral blood of 35 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD56+,CD19+ lymphocytes and CD4+/CD8+.Results The percentages of CD3+ T lymphocyte (61.58 ± 6.45 ) %,CD4+ T lymphocyte (28.38 ±4.89)% and the ratio of CD4+/CD8+ 0.99 0.22 in patients with ITP were lower than those in healthy controls[( 67.85 ± 4.68 ) %,( 38.00 ± 3.37 ) %,1.54 ± 0.13,all P ＜ 0.05].After immunosuppressive therapy,the percentages of CD3+ T lymphocyte ( 69.41 ± 5.03 ) %,CD4+ T lymphocyte (38.17 ±3.18)% and the ratio of CD4+/CD8+ 1.60 ±0.15 recovered to control levels.The percentages of CD8+ T lymphocyte (29.20 ±4.50)% and CD19+B lymphocyte ( 17.74 ±4.14)% were higher than those in healthy controls[( 24.82 ± 2.93 ) % and ( 12.09 ± 3.51 ) %,all P ＜ 0.05].After the immunosuppressive therapy,the percentages of CD8+ T lymphocyte ( 24.06 ± 3.02 ) % and CD19+ B lymphocyte ( 10.90 ± 3.55 ) %recovered to control levels.There were no significant difference of the percentage of CD56+ lymphocyte among ITP patients ( 15.80 ± 2.85 )%,ITP patients after immunosuppressive therapy ( 15.16 ± 2.77 )% and healthy controls ( 16.36 ± 2.75 ) %.ConclusionThe aberrant immunocyte subsets are involved in the pathogenesis of ITP,and detection of immunocyte subsets might be helpful for the diagnosis and determination of therapeutic outcome of ITP.

Insulin-like growth factor-1 receptor (IGF-1 R) widely exists in the surface of various types of cells and is closely associated with the formation and development of tumor cells.It also provides a new direction for the targeted therapy for tumors.This article reviews the expression,development,and progression of IGF-1R in pancreatic cancer and research advances in IGF-1R as a target for tumor treatment.

OBJECTIVE To evaluate the possibility that CdCl2 induces autophagy and apoptosis in HEK293 cells,and the role of extracellular regulated protein kinases (ERK1/2) and AKT proteins in autophagy. METHODS Green fluorescence protein(GFP)-light chain 3B(LC3B)expression plasmid was transfected into HEK293 cells. After 24 h,HEK293 cells were induced with CdCl2 2,4,8 and 10μmol·L-1 for 12 h. The expression of GFP-LC3B was detected by fluorescent microscopy. HEK293 cells were induced with CdCl2 2,4,8 and 10μmol · L-1 without transfection of GFP-LC3B for 12 h while autophagic vacuoles were observed by transmission electron microscopy. The expression of LC3B-Ⅱ/Ⅰproteins and the phosphorylation levels of ERK1/2 and AKT were analyzed by Western blotting. Apoptosis was detected by flow cytometry microscopy. HEK293 cells were treated with 3-MA 20μmol · L-1+CdCl2 10 μmol · L-1 for 12 h before cleaved caspase 3 protein was detected by Western blotting. RESULTS When HEK293 cells were exposed to CdCl2(≤10μmol · L-1)for 12 h,cytoplasmic GFP-LC3B punctuates were observed under the fluorescence microscope,and autophagic vacuoles were observed under an electron microscope. The expression of LC3B-Ⅱ/Ⅰ,p-ERK1/2 and p-AKT proteins was significantly increased in CdCl2-induced cells(P<0.05,P<0.01). Moreover,apoptosis was observed. The addition of 3-MA 20μmol · L-1+CdCl2 10μmol · L-1 enhanced apoptosis. Cleaved capase 3 protein expression was significantly increased(P<0.01). CONCLUSION CdCl2(≤10μmol·L-1)can induce autophagy in HEK293 cells. ERK1/2 and AKT proteins might be associated with the activation of autophagy that is accompanied by apoptosis,suggesting that autophagy can inhibit apoptosis at certain concentrations of CdCl2.

Background The biomeasurement and imageology of retinal nerve fiber layer(RNFL) thickness showed the damage of retinal structure in the early and middle stage of glaucomatous eye,however,the subtle functional damage of glaucoma can not be timely reflected only with automatic static perimeter.Microperimetry is a method of quantitatively assessing mean sensitivity (MS) of macular zone.Objective This study was to evaluate and compare the macular functional change in early and middle stage of primary open angle glaucoma (POAG) and chronic angle-closure glaucoma (CACG) with MP-1 microperimeter.Metbods This trail protocol was approved by Ethic Committee of Peiking University Third Hospital,and written informed consent was obtained from each subject prior to entering the study group.A cross-sectional and case-controlled study was designed.A total of 126 eyes from 126 subjects were enrolled in the trail,including 53 eyes of 53 normal subjects,50 eyes of POAG patients and 23 eyes of CACG patients.A macular 10° program was set with MP-1 microperimetry to record the MS of various subareas.The macula was zoned into central 2°,6°,and 10° visual fields and 4 quadrants in each ring.The MS of different rings and subareas was detected and compared among POAG patients,CACG patients and normal controls.Results The MS values of central 2°,6°,10° and whole macular area were (15.09 ± 3.03),(15.72 ± 3.22),(13.99 ± 3.63) and (14.91±3.07)dB in the POAG group,which were significantly lower than those of (17.29±1.59),(18.05±1.24),(16.76±1.89) and (17.37±1.46)dB in the normal control group (all at P=0.000).The MS values of central 2°,6°,10° and whole macular area was (16.00±2.39),(15.83±2.63),(14.45±3.15) and (15.42±2.54) dB in the CACG group,and the reduced MSs were seen at the 6°,10° rings and whole macular area in the CACG group compared with the normal control group (P =0.004,0.013,0.011).Within the 6° ring,the MS values in the inferotemporal quadrant were declined in the POAG group and CACG group compared with the normal control group (P =0.000,0.022),but the difference was not statistically significant between the POAG group and the CACG group (P =0.311).In addition,the MS value in the inferonasal quadrant was significantly lower than that of the normal control group (P =0.005); while that in the CACG group was not significantly different in comparison with the normal control group (P=0.119).In the POAG group,the MS value of the inferotemporal quadrant was significantly lower than that of the superonasal or superotemopral quadrant (P =0.043,0.016),but no significant differences were found among the 4 quadrants in the CACG group (all at P＞0.05).Conclusions The mild damage of retinal function appears in the early and middle stage of POAG and CACG.More serious MS reducing occurs in the inferotemporal and inferonasal quadrants of POAG.

Background and purpose:Radiofrequency ablation (RFA) was one of the best combined treatments for hepatocarcinoma. CT was commonly used to evaluate treatment response and recurrent disease. However it was diffi cult to evaluate treatment effectiveness of RFA and to detect recurrent nodule especially the size below 1cm. Digital subtraction arteriography(DSA) may provide fi nal diagnosis of recurrent nodule after RFA for hepatocarcinoma. Our purpose was to analyze the signs of DSA after RFA for hepatocarcinoma in order to provide possible reference for treatment effectiveness and the imaging follow-up methods. Methods:17 patients with primary liver cancer (n=15) or hepatic metastasis(n=2) were enrolled in this study. Common hepatic arteriography or super-selective angiography of suspicious tumor area were performed on all patients. Results:On DSA, most ablated regions presented as round or ovoid low density areas with no stain. At the peripheral zone of the lesion, fi ve signs after RFA could be found: Stain of localized granulation tissue, arterio-portal fi stula, hemorrhage, recurrence and no abnormal fi ndings. Local or intrahepatic recurrence occurred in 9 cases. Conclusion:Hepatic DSA has great value in detecting local recurrence of hepatic tumor after RFA treatment. DSA is superior to CT in detecting marginal or intrahepatic recurrent nodule below 1 cm.

Objective To detect the doubling time of airway smooth muscle cells of human and rat by the xCELLigence in-strument,a real time cellular analyzer.Methods The airway smooth muscle cells were separated by collagenase-pancreatin digestion from the rat airway.Then,added to the different holes of E-plate of xCELLigence instrument with the planting destiny of 3 000 cells/well.and so were the human airway smooth muscle cells.The E-plate was then placed on the xCELLi-gence instrument to monitor cell proliferation for 100 hours to calculate the doubling time by using the RTCA Software Package 2.0 software.Results The doubling time of human airway smooth muscle cell calculated by the real time cellular analyzer was 23.96±0.47 h,which was consistent with the data provided by the reference.The doubling time of rat airway smooth muscle cell was 18.62±0.15 h,and the computational process was simple,time-saving and also effective.Conclusion The xCELLigence instrument can be used to calculate doubling time of airway smooth muscle cells of human and rat, which provides experimental methods and reference data for the basic respiratory disease research.

Objective To observe the efficacy of cumulative dose of polyethylene glycol-interferon (Peg-IFN)α-2a combined with ribavirin in patients with decompensated hepatitis C virus (HCV)-related liver cirrhosis , and to evaluate the effects of anti-virus therapy on the progress of the disease . Methods From January 2005 to March 2009 ,patients with decompensated HCV-related liver cirrhosis were enrolled ,also included patients received partial splenic embolization .Peg-IFNα-2a combined with ribavirin therapy was given to patients whose blood cell met interferon (IFN) therapy standards .The dosage of Peg-IFNα-2a and ribavirin was adjusted according to the tolerance of the patients .After the treatment ,the patients were followed-up for 24 weeks .The patients whose blood cell did not meet IFN therapy standards and the patients unwilling to receive anti-virus therapy were assigned to control group and were followed-up for 96 weeks .The total amount of medication was calculated according to cumulative exposure dose . Sustained virological response (SVR ) , recurrence rate , liver function and disease progression were observed .The t test or Chi-square test was performed for comparison between groups and rate of disease progression was analyzed with Kaplan Meier curve .Results After anti-virus therapy , SVRs of patients with cumulative dose of Peg-IFNα-2a and ribavirin over 60% (include 60% ) were 27 .3%(12/44) and 27 .7% (13/47) ,respectively ;the recurrence rates were 7/19 and 35 .0% (7/20) ,respectively . In patients with cumulative dose less than 60% ,SVRs were 1/7 and 1/4 ,respectively ,and the recurrence rates were both 1/2 ;the differences of different doses was not statistically significant (all P>0 .05) .At the 24th week of follow-up after therapy ,the Child-Pugh score of combined therapy group was 7 .9 ± 1 .4 , which was lower than that before treatment (8 .5 ± 1 .2) ,and the difference was statistically significant (t=2 .33 ,P=0 .02) .At the 96th week of follow-up ,the Child-Pugh score of control group was 10 .0 ± 1 .6 ,which was higher than that before treatment (8 .5 ± 1 .4) ,and the difference was statistically significant (t=5 .82 , P<0 .01) .The disease progression rate of combined therapy group was 15 .7% , which was lower than that of control group (32 .4% ) ,and the difference was statistically significant (χ2=4 .34 ,P= 0 .04) .Conclusion The application of non-standard dosage of Peg-IFNα-2a combined with ribavirin in the patients with decompensated HCV-related liver cirrhosis can achieve virological response once the cumulative dose reached certain standards ,improve Child-Pugh scores of patients and slow disease progression .

Background Penetrating keratoplasty (PKP) has become an effective method of treatment for fungal keratitis in recent years,but the application timing of glucocorticoids after PKP is still unclear.Literature reported that the concentration of tear (1,3)-β-D-glucan in fungal keratitis was significantly higher than that in normal.Objective This study was to investigate the change of tear (1,3)-β-D-glucan before and after PKP in fungal keratitis and to explore the application duration of anti-fungal drugs and application timing of glucocorticoids.Methods This study protocol was approved by ethic committee of Affiliated Hospital of Qingdao University.A serial cases-observational study was performed from August,2011 to December,2012.Twenty eyes of 20 patients with fungal keratitis were collected in Affiliated Hospital of Qingdao University.PKP was performed in affected eyes,and the fellow health eyes served as controls.Tear of 50 μl was obtained in the controls on 1 day before operation and 1 day,7,14,21 and 28 days after operation to detect tear (1,3)-β-D-glucan levels.Results Tear (1,3)-β-D-glucan levels were (14.67±3.84)mg/L,(1 861.66±196.17) mg/L,(927.71±155.82)mg/L,(392.30±71.22)mg/L,(179.60±40.47) mg/L,(40.20± 12.46) mg/L and (15.12± 1.80) mg/L in the control group,preoperative 1 day,postoperative 1 day,7,14,21,28 days,respectively,showing a significant difference among various time points (F=883.45,P=0.00).Tear (1,3)-β3-D-glucan levels were gradually reduced with the lapse of the postoperative time,with significant differences between adjacent timepoints (t' =13.84,t =16.67,t' =11.02,t' =13.97,t' =-8.45,all at P=0.00).Tear (1,3)-β-D-glucan levels in postoperative 28 days came near that of normal control group,without significant difference between them (P =0.64).Fungal keratitis recurred in 2 eyes on the fifth and sixth day after operation,with the tear (1,3)-β-D-glucan levels of 2 350.24 mg/L and 1 992.82 mg/L,respectively.Conclusions The concentration of (1,3)-β-D-glucan in the tears increases in the eyes with fungal keratitis and drops to normal range at 28 days after PKP,indicating that the antifungal eyedrops should be applied until 4 weeks after PKP,and this is an optimal timing of using corticosteroid eyedrops to resist reject reaction.

Objective:To study the expression of miRNA-7 in B lymphocytes of primary Sj?gren's syndrome (pSS) and its relationship with phosphatase andtensin homolog deleted (PETN) and disease activity.Methods:Twenty newly diagnosed outpatient and inpatient pSS patients were used as case group collected from January 2017 to December 2019 of Qinghai Provincial People's Hospital. Twentyhealthy persons were used as the control group. Disease-related indicators of the case group were collected. Quantitative polymerase chain reaction (RT-qPCR)was used to detect miRNA-7 and PETN mRNA expression in B lymphocytes of the two groups and the consistency between miRNA-7 expression in the plasma and B lymphocytes of the case group was analyzed. Western Blotting method was used to detect the PETN protein in B lymphocytes of the two groups. Correlation analysis was used to analyze the relationship between miRNA-7 expression in B lymphocytes and disease activity in the case group. Linear regression analysis was performed between miRNA-7 and PETN mRNA.Results:The expression of miRNA-7 (0.53±0.17) in the B cells increased and the expression of PTEN mRNA (0.88± 0.24) and protein (0.51±0.12) in the case group were reduced compared with that of miRNA-7(0.39±0.11), PTEN mRNA(2.32±0.30) and protein(1.03±0.21) of the control group. The above differences were statistically significant ( t=2.990, P<0.05; t=16.98, P<0.05; t=8.41, P<0.05). Linear regression showedthat PTEN miRNAwas negatively correlated with miRNA-7 ( b=-0.78, P<0.01), the expression of miRNA-7 in the case group was positively related with EULAR Sj?gren′s syndrome Disease Activity Index (ESSDAI), IgG, IgA, anti-SSB and was negatively correlated with C4 and WBC. Conclusion:There is a certain relationship between miRNA-7 and disease activity. MiRNA-7 may participate in the pathogenesis of pSS byregulating PETN in B cells of pSS. MiRNA-7 has certain clinical value for disease activity evaluation.

Objective To evaluate the effect of midwives′antenatal clinic service on management of gestational weight and birth outcomes .Methods Totally 420 primiparas who had made registration and regular antenatal examination in a hospital ,were randomly divided into two groups .The women in the control group(n=208) received routine antenatal clinicservice .In addition ,the women in the experimental group(n=212) participated in an individual midwives′antenatal clinic .The primiparas′gestational weight gain and birth outcomes were measured and compared between the two groups .Results The rate of natural birth in the experimen‐tal group was higher than that of the control group .The rate of fetal macrosomia ,episiotomy and labor analgesia were lower than that of the control group(P<0 .01) .The rate of naturalbirth ,fetal macrosomia ,episiotomy ,labor analgesia were significantly lower than that of the control group(P<0 .01) .The neonatal weight[(3 250 .58 ± 270 .03)g vs .(3 405 .42 ± 403 .28)g] and volume of bleeding[(199 .13 ± 64 .37)mL vs .(249 .53 ± 93 .56)mL] 2 hours postpartum in the experimental group were significantly lower than that of the control group(t=28 .610 ,P<0 .01 ;t=20 .581 ,P<0 .01) .There were no significant differences in the rate of gesta‐tional diabetes mellitus ,pregnancy‐induced hypertension and fetal distress in uterusbetween the two groups (P>0 .05) .Conclusion The midwives′antenatal clinic service has good effects on management ofgestational weight gain ,improves birth outcomes and pro‐vides primiparas positive experience of childbirth .