ObjectiveTo investigate the expression and significance of CD28- cells, CD4+ and CD8+T lymphocytes in the peripheral blood and synovial fluid in patients with rheumatoid arthritis ( RA ). Methods The expression of CD28, CD4+, CD8+ T lymphocytes and inducible co-stimulator(ICOS) in the peripheral blood and synovial fluid in 45 patients with RA were detected by three-color flow cytometry. Independent sample's t test was used for statistical analysis between the two groups. ResultsSynovial fluid CD4+CD28+ICOS+, CD4+CD28- ICOS+ , CD8 + CD28 + , CD8 + CD28 + 1COS+ T lymphocytes were significantly increased than the peripheral blood in RA patients[(36±19)％ vs (15±8)％, t=-4.234, P＜0.01; (2.1±2.2)％ vs (0.6±1.4)％, t=-3.143, P＜0.01; (62±15)％ vs (47±18)％, t=-2.885, P＜0.01; (9±9)％ vs (3±3)％,t=-2.131, P＜0.05], Synovial fluid CD8+CD28-T lymphoc-ytes were significantly reduced than the peripheral blood[(38±15)％ vs (54±18)％, t=2.975, P＜0.01], Synovial fluid CD8+ CD28-ICOS+, CD4+CD28+and CD4+ CD28- T lymphocytes had no significant difference than the peripheral blood (P＞0.05). Compared with peripheral blood in the same patients with RA, CD4+CD28+ ICOS+, CD8+ CD28+ T lymphocyteswere significantly increased[(38±18)％ vs (16±10)％, t=-4.065, P＜0.01 ; (61±16)％ vs (41±21)％, t=-4.065,P＜0.01], CD8+CD28-T was significantly reduced[(39±16)％ vs (59±21)％, t=2.949, P＜0.01]. The level of CD4+ CD28-, CD8+ CD28-, CD28-ICOS+ T lymphocytes in the active and remission patients with RA was not significantly different (P＞0.05). ConclusionSynovial fluid CD28T lymphocyte subsets disturbance and the abnormal expression of ICOS in patients with RA may play important roles in the mechanism of joint damage.

Objective To evaluate the efficacy and safety of different bismuth-based quadruple therapies for Helicobacter pylori (H.pylori) eradication.Methods From December 2012 to October 2013,240 patients with H.pylori infection were collected and evenly divided into clarithromycin group,levofloxacin group,furanzolidone group and metronidazole group.Each group received bismuth potassium citrate 220 mg,esomeprazole 20 mg and amoxicillin 1 000 mg twice daily.In addition,each group received clarithromycin 500 mg,levofloxacin 200 mg,furanzolidone 100 mg,and metronidazole 400 mg,respectively.The course of treatment was 10 days.At least four weeks after the end of therapy and withdrawal the medicine,patients underwent fasting 13C-urea breath test or 14C-urea breath test.The negative result indicated as successful H.pylori eradication.The adverse effects were observed and recorded during treatment.The rate of H.pylori eradication was analyzed by the intention to treat (ITT) analysis and per protocol (PP) analysis.Chi-square test was performed for eradication rate comparison among groups.Results According to ITT analysis,the eradication rate of clarithromycin group,levofloxacin group,furanzolidone group and metronidazole group was 81.67％ (49/60),88.33％ (53/60),93.33％ (56/60) and 73.33％ (44/60),respectively,and according to PP analysis which was 85.96％ (49/57),89.83％ (53/59),94.92％ (56/59) and 75.86％ (44/58),respectively.The differences among four groups were statistically significant (x2 ＝10.13 and 9.89,both P＜0.05).The differences between furanzolidone group and metronidazole group were statistically significant (x2 =8.64 and 8.55,both P＜0.01).There were no statisticaly significant differences in adverse effects among the four groups (x2 =0.47,P＞0.05).Conclusion The H.pylori eradication rate is high in furanzolidone contained bismuth based quadruple therapy and with good safety,which could be the first line treatment for H.pylori eradication.

Objective To investigate the correlation between the drug resistance of Helicobacter pylori (H .pylori )and clinical eradication efficacy of bismuth-based quadruple therapies,and to guide clinical rational drug use in the region.Methods A total of 260 patients with H .pylori infections were collected.H .pylori from biopsied gastric mucosa tissues were isolated and cultured.Drug resistant rates of isolated H .pylori to metronidazole,clarithromycin,amoxicillin,levofloxacin and furanzolidone were tested.Patients were randomly divided into clarithromycin,levofloxacin,furanzolidone and metronidazole groups by completely randomized design.All patients received bismuth potassium 220 mg,esomeprazole 20 mg and amoxicillin 1 000 mg twice daily,and according to group received clarithromycin 500 mg, levofloxacin 200 mg,furanzolidone 100 mg and metronidazole 400 mg,twice daily,espectively.The treatment course was 10 days.At least four weeks after treatment,13 Curea breath test or 14 Curea breath test was taken.According to the intention to treat (ITT)and per-protocal (PP),the eradication rate of each group was caculated.Chi square test was performed to compare the differences between groups. Results The drug resistant rate of H .pylori to metronidazole,clarithromycin,amoxicillin,levofloxacin and furanzolidone was 94.2% (146/155 ), 21 .3% (33/155 ), 2.6% (4/155 ), 5 .8% (9/155 ) and 1 .9%(3/155),respectively.According to ITT analysis,the eradication rate of clarithromycin group, levofloxacin group,furanzolidone group and metronidazole group was 81 .5 %(53/65 ),90.8%(59/65 ), 93.8% (61/65 )and 75 .4%(49/65),respectively.And according to PP analysis which was 84.1 %(53/63),92.2%(59/64),95 .3%(61/64)and 79.0%(49/62 ),respectively.The differences between furanzolidone group and metronidazole group,clarithromycin group were staistcally significant (χ2ITT =8.509 and 4.561 ;χ2PP = 7.592 and 4.323,all P < 0.05 ).There was no statistical significance in the H .pylori eradication rate between resistant strains and sensitive strains of each group.Conclusion Bismuth-based quadruple therapy can overcome antibiotic resistance,the eradication rate of protocal with furanzolidone is higher and with good safety,which can be the first-line treatment for H .pylori eradication.

Objective To investigate the effect of dexmedetomidine combined with butorphanol conventional therapy on sustaining sedation of intensive care unit (ICU) patients.Methods Sixty critically ill patients in Binzhou Central Hospital from June to September in 2014 were randomly divided into experimental group and control group, 30 cases in each group. In the control group, 0.8 mg/L dexmedetomidine hydrochloride injection (400μg with addition of 46 mL normal saline to form 50 mL solution) was intravenously infused continuously at a speed of 0.4μg·kg-1·h-1 by a micro-pump to induce analgesia and sedation; while in the experimental group, dexmedetomidine combined with 200 mg/L butorphanol (10 mg plus 40 mL normal saline to form 50 mL solution) was given for intravenous infusion by a micro-pump with a speed of 0.01 mg·kg-1·h-1 to maintain analgesia and sedation for 48 hours whose required Ramsay score in both groups was 3 - 5. Before and after treatment, the changes of heart rate (HR), respiratory rate (RR), mean arterial pressure (MAP), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), and the pulse oxygen saturation (SpO2) of both groups were observed. The dosage of dexmedetomidine used for maintenance of required analgesia and sedation and FPS (facial expression) grading and Ramsay score were compared respectively between the two groups, and the clinical efficacy of the two groups were evaluated.Results After treatment, the HR, MAP, RR in both groups were significantly lower, and PaO2 and SpO2 were significantly higher than those before treatment, and the degrees of improvement in the above indexes of the experiment group were superior to those of the control group [HR (bpm): 84.58±12.43 vs. 118.62±14.21, MAP (mmHg, 1 mmHg = 0.133 kPa): 82.35±12.12 vs. 92.35±12.32, RR (times/min): 25.42±3.98 vs. 32.87±5.12, PaO2 (mmHg): 95.21±10.55 vs. 75.18±8.57, SpO2: 0.981 4±0.102 8 vs. 0.954 7±0.093 8, allP < 0.05]. The total therapeutic effect in experiment group was significantly higher than that in control group [93.3% (28/30) vs. 76.7% (22/30),P < 0.05]. The dexmedetomidine dosage used in the experiment group was much less than that in the control group (μg/d: 412.12±23.18 vs. 520.05±15.68,P < 0.05). The FPS score in the experiment group was obviously lower than that in the control group (1.48±0.16 vs. 2.52±0.74,P < 0.05).Conclusion In comparison, to achieve sustained and required analgesic and sedative effect for ICU patients by combined use of dexmedetomidine and butorphanol, the dosage of dexmedetomidine used is less than dexmedetomidine applied alone, in addition, the combined use can achieve better Ramsay grading, steady blood pressure and excellent effect.

Objective:To analyze the applicability and feasibility of a cell-free DNA barcode-enabled single-molecule test (cfBEST) in non-invasive prenatal diagnosis of phenylketonuria.Methods:This study recruited four pregnant women who were prenatally diagnosed as heterozygous carriers of hot spot mutations in the PAH gene from pedigrees with phenylketonuria at the First Affiliated Hospital of Zhengzhou University from July to September 2019. The frequency of mutations in maternal plasma cell-free DNA and the fetuses' genotypes were analyzed by cfBEST. Nested polymerase chain reaction primers were designed to amplify the mutation sites in each pedigree. The results of cfBEST were compared with those of invasive prenatal diagnosis. Descriptive analysis was used for data analysis. Results:In pedigree 1, the frequency of c.603T>G and c.842+2T>A mutations in maternal plasma cell-free DNA were 48.40% (291/601) and 9.70% (61/628), which was detected by cfBEST. The fetus was diagnosed with phenylketonuria with two heterozygous mutations. In pedigree 2, the frequency of c.1238G>C and c.842+2T>A mutations in maternal plasma cell-free DNA was 43.70% (786/1 798) and 0% (0/1 550), respectively. Both mutations were wild-type, and the fetus was neither phenylketonuria nor a carrier. In pedigree 3, the frequency of c.1045T>G and c.728G>A mutations in maternal plasma cell-free DNA was 44.00% (930/2 112) and 0% (0/705), respectively, suggesting that both mutations in the fetus were wild-type, and the fetus was neither phenylketonuria nor a carrier. In pedigree 4, the frequency of c.755G>A and c.728G>A mutations were 45.40% (743/1 637) and 4.50% (28/849), respectively, which indicated that the former was wild-type, and the latter was heterozygous; namely the fetus was a carrier of phenylketonuria. The results of cfBEST were consistent with those of invasive prenatal diagnosis. Three pedigrees (Pedigree 2, 3 and 4) continued the pregnancy to full-term, and the phenylalanine levels in the neonates were all below 120 μmol/L. No abnormalities were reported in those three infants during follow-ups at one, three, and six months after birth.Conclusions:The cfBEST could be used for non-invasive prenatal diagnosis of phenylketonuria caused by PAH gene mutation, but further studies with a larger sample size are needed.

Turnner syndrome is a common sex chromosome disorder. We reported a rare case with Turnner syndrome caused by abnormal number and structure of sex chromosomes. Hereby fluorescence in situ hybridization (FISH) and copy number variation by whole genome low depth sequencing (CNV-seq) were used to clarify the abnormal chromosome. This study provides a diagnostic strategy for clinicians and genetic researchers.

OBJECTIVETo compare the clinical efficacy of Chinese traditional herbs (CTH) alone or combined with Western medicine (conventional treatment plus different doses of prednisone) in treating anaphylactic purpura nephritis (APN).

METHODSClinical data of 232 patients with APN were collected and analyzed. They were assigned to four groups. Patients in Group A1 were treated with CTH; in Group A2 were treated initially with Western medicine but turned midway to CTH; in Group B1 and B2 treated with CTH combined with conventional Western medicine and plus low or high dose of prednisone respectively. The comprehensive clinical efficacy on symptoms, physical signs, routine urine examination, blood creatinine and urea nitrogen, as well as the treatment duration and long-term effect in the four groups were observed.

RESULTSThere was no significant difference in the comprehensive clinical effects among the four groups (P > 0.05). However, the treatment duration was significantly shorter in Group A1 and B1 than in the other two groups (P < 0.01). Follow-up study on patients in similar duration showed similar reoccurrence rate among groups (P > 0.05).

CONCLUSIONChinese herbs has definite effects on APN with the treatment course shorter than that of other treatments, the optimal protocol for treatment of APN is applying Chinese herbs alone or combined with conventional Western medicine plus low dose prednisone. It is necessary to conduct a follow-up study even though the patients have been cured.

Adolescent ; Adult ; Anti-Inflammatory Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Nephritis ; drug therapy ; etiology ; Phytotherapy ; Prednisone ; therapeutic use ; Purpura, Schoenlein-Henoch ; complications ; drug therapy ; Treatment Outcome ; Young Adult

Oxidative stress could induce apoptosis and autophagy process simultaneously, but the role of autophagy is still not clear. Beclin 1, a key gene regulating the preautophagosome formation, is involved in the injury induced by oxidative stress. To observe the role of autophagy in H2O2-induced injury of U251 cells, the recombinant plasmid Psilencer3.1-siRNA-Beclin 1 was transfected into U251 cells by eukaryotic cell transfection technique. Plasmid vector and cell culture medium were used as negative and control groups respectively. The cells were collected 24 h later, and the cell total protein was extracted to detect Beclin 1, Bcl-2 and Bax protein expressions by Western blot. After the Beclin 1-siRNA cells were treated with 1 mmol/L H2O2, the autophagic vacuoles in the cells were stained with monodansylcadaverine (MDC), and the cell apoptotic ratio was determined with PI/Annexin V-FITC staining by flow cytometry analysis. The results showed that the synthetic siRNA decreased the expression of Beclin 1 protein significantly, but had no obvious effect on the levels of Bcl-2 and Bax protein expressions. Compared with those in the control group, the autophagic vacuoles, the level of LC3-II protein expression and the percentage of apoptotic cells increased (P < 0.05) in 1 mmol/L H2O2 group. In Beclin 1-siRNA + H2O2 group, autophagic vacuoles and the levels of LC3-II protein expression decreased obviously, the percentage of apoptotic cells increased significantly compared with that in 1 mmol/L H2O2 group (P < 0.05). H2O2 and autophagy inhibitor 3-methyladenine (3-MA) combination also increased the percentage of apoptotic cells obviously (P < 0.05). These results revealed that the transfection of Psilencer3.1-siRNA-Beclin 1 effectively inhibited the expression of Beclin 1 protein expression, degraded the autophagy level and increased the apoptotic rate in U251 cells under oxidative stress, which was coincident with the effect of autophagy inhibitor 3-MA. This study suggests that autophagy is a cell protective role in oxidative stress process, and the inhibition of autophagy may enhance apoptosis.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Autophagy ; physiology ; Beclin-1 ; Brain Neoplasms ; pathology ; Cell Line, Tumor ; Glioma ; pathology ; Humans ; Hydrogen Peroxide ; pharmacology ; Membrane Proteins ; genetics ; metabolism ; Oxidative Stress ; RNA, Small Interfering ; genetics ; Transfection

Objective: To discuss the correlation between aerobic exercise ability, pulmonary function and heart rate recovery ability of 7-12 year-old students under different altitudes, so as to provide evidence on the physiological changes of exercise and characteristics of altitude training among children and adolescents at different altitudes. Methods: A total of 235 students aged 7-12 years old living at 3 240 m(Hi group), 300 students at 1 290 m(SubHi), and 300 students living between 1 290 m and 3 240 m (HiSubHi) were selected to perform 20-mSRT test, respiratory function test and heart rate recovery ability evaluation after quantitative load exercise. Results: Performance on 20-mSRT, FVC, FEV1, FEV1% and V% of students aged 7-12 years at different altitudes showed an upward trend with the increase of age. After adjusting for altitudes, there were significant differences in average level of these indicators. Average level of 20-mSRT, FVC, FEV1%, FEV1% and V% in SubHi group were significantly higher than those in Hi group(P<0.05), and the HiSubHi group showed an upward trend compared with Hi group after 1-year adaptation to the sub-highland environment; Except for male FVC (r=0.33, P=0.24) and FEV1 (r=0.34, P=0.22) in HiSubHi group, and the male FEV1% (r=0.46, P=0.14) in SubHi group, all the indicators showed significant association with 20-mSRT(P<0.05); 20-mSRT was only significantly correlated with heart rate recovery ability among SubHi group (male: r=0.78, P<0.05); Female(r=0.81, P=0.01), there was no significant correlation between 20-mSRT and heart rate recovery ability in other groups(P>0.05). Conclusion The results suggest developmental characteristics of aerobic exercise ability, lung function and heart rate recovery ability among 7-12 years old students under different altitudes. The correlation among 20-mSRT, FVC, FEV1, FEV1% and V% might involve high altitude adaptation.