Objective To investigate the effect of recombinant Schistosoma japonicum egg ribonuclease SjCP1412 (rSjCP1412) on proliferation, cell cycle, apoptosis and activation of human hepatic stellate cells LX-2 in vitro, and explore the underlying mechanisms. Methods The rSjCP1412 protein was expressed in Escherichia coli BL21 by prokaryotic expression, and the highly purified soluble rSjCP1412 protein was prepared by Ni NTA affinity chromatography and urea gradient refolding dialysis. Yeast RNA was digested using 12.5, 25.0, 50.0 µg rSjCP1412 proteins at 37 °C for 2, 3, 4 h, and the enzymatic products were electrophoresed on 1.5% agarose gel to observe the RNAase activity of rSjCP1412 protein. The proliferation of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was measured using CCK-8 assay, and the apoptosis of LX-2 cells stimulated by different doses of rSjCP1412 protein for 48 hours was detected using the Annexin V-FITC/PI double staining, while the percentage of LX-2 cells at G0/G1, S and G2/M phases of cell cycle following stimulation with different doses of rSjCP1412 protein for 48 h was detected by DAPI staining. The type I collagen, type III collagen and α-smooth muscle actin (α-SMA) mRNA expression was quantified using quantitative florescent real-time PCR (qPCR) assay and Western blotting at transcriptional and translational levels in LX-2 cells following stimulation with different doses of rSjCP1412 protein for 48 h, while soluble egg antigen (SEA) served a positive control and PBS without rSjCP1412 protein as a normal control in the above experiments. The expression of collagen I, α-SMA and Smad4 protein was determined using Western blotting in LX-2 cells following stimulation with rSjCP1412 protein, transforming growth factor-β1 (TGF-β1) alone or in combination, to examine the signaling for the effect of rSjCP1412 protein on LX-2 cells. Results The rSjCP1412 protein was successfully expressed and the highly purified soluble rSjCP1412 protein was prepared, which had a RNase activity. Compared with the normal group, the survival rates of LX-2 cells significantly decreased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein and SEA for 48 h (F = 22.417 and 20.448, both P values < 0.05). The apoptotic rates of LX-2 cells significantly increased post-treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h (F = 11.350, P < 0.05), and treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h resulted in arrest of LX-2 cells in G0/G1 phase (F = 20.710, P < 0.05). Treatment with 12.5, 25.0, 50.0 µg/mL rSjCP1412 protein for 48 h caused a significant reduction in relative expression levels of collagen I (F = 11.340, P < 0.05), collagen III (F = 456.600, P < 0.05) and α-SMA mRNA (F = 23.100, P < 0.05) in LX-2 cells, and both rSjCP1412 protein and SEA treatment caused a significant reduction in collagen I (F = 1 302.000, P < 0.05), α-SMA (F = 49.750, P < 0.05) and Smad4 protein expression (F = 52.420, P < 0.05) in LX-2 cells. In addition, rSjCP1412 protein treatment inhibited collagen I (F = 66.290, P < 0.05), α-SMA (F = 31.300, P < 0.05) and Smad4 protein expression (F = 27.010, P < 0.05) in LX-2 cells activated by TGF-β1. Conclusion rSjCP1412 protein may induce apoptosis of LX-2 cells and inhibit proliferation, cell cycle and activation of LX-2 cells through down-regulating Smad4 signaling molecules.

Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.

Objective:To compare the incidence of coronary microvascular disease (CMVD) between patients with non-obstructive and obstructive coronary arteries.Methods:We retrospectively analyzed 97 patients with angina pectoris, who underwent the absolute quantitative PET examination of myocardial perfusion and coronary anatomy examination within 90 days. All patients were divided into two groups: non-obstructive group (72 cases, no stenosis ≥50% in all three coronary arteries) and obstructive group (25 cases, at least one coronary stenosis ≥50%; and at least one coronary stenosis<50%). Quantitative parameters derived from PET including rest myocardial blood flow (RMBF), stress myocardial blood flow (SMBF), coronary flow reserve (CFR) and cardiovascular risk factors were compared between the two groups. CMVD was defined as CFR<2.90 and SMBF <2.17 ml·min -1·g -1. Results:Incidence of CMVD was significant higher in the non-obstructive coronary arteries of the obstructive group than in the non-obstructive coronary arteries of non-obstructive group (47.1% (16/34) vs. 25.5% (55/216), χ 2=6.738, P=0.009) while incidence of CMVD was similar between non-obstructive and obstructive patients ((44% (11/25) vs. 33.3% (24/72), χ 2=0.915, P=0.339). RMBF ((0.83±0.14) ml·min -1·g -1 vs. (0.82±0.17) ml·min -1·g -1), SMBF ((2.13±0.60) ml·min -1·g -1 vs. (1.91±0.50) ml·min -1·g -1) and CFR (2.59±0.66 vs. 2.36±0.47) were similar between the two groups (all P>0.05). Conclusions:CMVD can occur in non-obstructive coronary arteries in both patients with non-occlusive coronary arteries and patients with obstructive coronary arteries. Prevalence of CMVD is significantly higher in patients with obstructive coronary arteries than in patients with non-obstructive coronary arteries. The CMVD severity is similar between the two groups.

Objective Toinvestigatethecharacteristicsofclinicalpathologyand MRIofintracranialsolitaryfibroustumor/hemangiopericytoma (SFT/HPC).Methods ThisstudyanalyzedtheMRIimages,pathologicalandclinicaldataof14SFT/HPCpatientsretrospectively. AllthecasesweresubjectedtoMRIplainscanandenhancementexamination.CharacteristicsofMRIofallcaseswerereviewedtogetherwith clinicopathologicchanges.Results AllSFT/HPClesionswerelocatedintheintracranialbutextra-cerebralspace.6werelocatedabovethe tentoriumofcerebellum,and2werelocatedbelowit.Lesionsof6patientswereacrossthetentoriumofcerebellumandspreadfrom supratentorialtosubtenorialspace.Amongalllesions,4wereroundinshape,10werelobulated,and3weresmallnodulesaroundthe edge.9ofthemexhibitednecrosisandcysticstructures.11lesionsshowedhypointensityand3casesshowedisointensityonT1WI.All thelesionswereheterogeneoushyperintensetyonT2WI,and5ofthemdisplayed"yin-yang"patternonT2WI.11casesexhibitededema.Signalof vascularvoidflow wasobservedin6cases.Thesolidpartsofthetumorsshowedsignificanthomogeneousenhancementon MRI. StrongpositiveSTAT6stainingwasobservedforthenuclearoftumorcells.Conclusion ItisdifficulttodifferentiateSFT/HPCfrom meningeoma.The"yin-yang"patternonT2WIisthecharacteristicofSFT/HPC.Inaddition,nuclearpositivestainingofSTAT6isalsospecific featureofSFT/HPCcell.

Objective To evaluate the prognostic value of peak ejection rate (PER) and peak filling rate (PFR) in patients with left ventricular systolic dysfunction (LVSD) after acute myocardial infraction (AMI).Methods A total of 123 patients (103 males,20 females,age:(60.6± 11.2) years) with LVSD after AMI who underwent 99Tc-methoxyisobutylisonitnle (MIBI) gated SPECT myocardial perfusion imaging (GSMPI) from January 2014 to December 2015 were retrospectively analyzed.Summed rest score (SRS) and total perfusion deficit (TPD) were acquired by using quantitative perfusion SPECT (QPS) soft-ware.Left ventricular ejection fraction (LVEF),end-diastolic volume (EDV),end-systolic volume (ESV),PER,and PFR were calculated by using quantitative gated SPECT (QGS) software.The clinical parameters of patients were recorded and the cardiac events were taken as the endpoint of follow-up (median time:27 (range:9-50) months).Pearson correlation was used to analyze the correlation between PER and PFR.Receiver operating characteristic (ROC) curve was used to evaluate optimal cut-off values of PER and PFR for predicting cardiac events.Kaplan-Meier survival analysis and Cox proportional hazards model were also used for data analysis.Results There was a great correlation between PER and PFR (r =-0.931,P＜0.001).Optimal cut-off values of PER and PFR for predicting cardiac events were-1.10 EDV/s and 1.09 EDV/s respectively.Kaplan-Meier survival analysis showed that cumulative survival rate without cardiac events was lower in patients (n=48) with-PER≤ 1.10 EDV/s than that in patients (n=75) with-PER＞ 1.10 EDV/s (16.7％ vs 66.7％;x2=60.096,P＜0.001),and the same rate in patients (n=50) with PFR ≤ 1.09 EDV/s was lower than that in patients (n=73) with PFR＞1.09 EDV/s (16.0％ vs 68.5％;x2=74.771,P＜0.001).Cox multivariate analysis showed that PER (hazard ratio (HR)=0.40,95％ CI:0.20-0.83) and PFR (HR=0.22,95％ CI:0.12-0.47) were independent predictors for cardiac events.Conclusion There is a great correlation between PER and PFR in patients with LVSD after AMI and they are independent predictors for cardiac events.

Objective To analyze the pulse-field gel electrophoresis (PFGE) patterns and antibiotic resistance of Salmonella enteritidisstrains isolated in Longyan city in order to provide reference for effective control, clinical diagnosis and treatment of outbreaks of food-borne diseases.Methods Seventy-seven Salmonella enteritidis strains isolated from food poisoning cases, livestock and poultry meat samples collected for food safety risk monitoring and sporadic cases in 2014 and 2016 were analyzed by PFGE.Antibiotic susceptibility testing was carried out to determine the minimum inhibitory concentrations (MIC) of 16 common antibiotics to 57 strains.Results Seventeen PFGE types were identified among the 77 Salmonella strains including three predominant types (P4, P6 and P8) and were grouped into three major pulsotypes (Cluster1-3) at 90% similarity level.The predominant types causing outbreaks of Salmonella enteritidis infection were P8, P6, P4 and P14, which were responsible for five cases of food poisoning.These isolated strains were highly resistant to nalidixic acid (93%), followed by resistance towards ampicillin (74%) and streptomycin (54%).Forty-one strains were resistant to three or more antibiotics (71.93%).Conclusion The PFGE types of Salmonella enteritidis strains isolated in Longyan city exhibit polymorphism and are clustered into three major pulsotypes.PFGE molecular typing can provide early-warning of sporadic outbreaks and help to identify the sources of infections in Salmonella enteritidis-related food poisoning.Attentions should be paid to multidrug resistant Salmonella enteritidis.Surveillance for multidrug resistant strains and usage of antibiotics should be strengthened.

Objective To compare the genotypes distribution of human papillomavirus (HPV ) infection in tissues of cervical cancers and cervical intraepithelial neoplasias (CIN ) and its clinical significance .Methods The polymerase chain reaction (PCR) and the gene-chips technique were utilized for the detection of 23 kinds of HPV genotypes in the tissue specimens from 192 cases of cervical intraepithelial neoplasia (CIN) and 85 cases of cervical cancers .And the related data of all subjects were analyzed .Results In 192 cases of CIN ,the total positive rate of HPV was 82 .29% (158/192) ,the positive rate of single genotype infection was 46 .88% (90/192) and the positive rate of multiple genotypes infection was 35 .42% (68/192);In 85 cases of cervical cancers ,the to-tal infection rate of HPV was 88 .24% (75/85) ,the positive rate of single genotype infection was 65 .88% (56/85) and the positive rate of multiple genotypes infection was 22 .35% (19/85) .Conclusion PCR combined with the gene-chips technique can be used in the detection of the tissue samples of cervical lesions ,once detection can detect 23 kinds of HPV genotypes with high sensitivity and strong specificity ,which has very important significance to the prevention and treatment of cervical cancer and precancerous lesions and the their vaccine research .

Objective To understand the current situation of occupational exposure and occupational safety attitudes among nursing students in Kunming Medical University, and to find effective interventions to improve nurses' occupational safety,and to reduce occupational exposure.Methods 854 students in Kunming Medical University were investigated by a questionnaire survey, including Demographic characteristics, Occupational exposure conditions, Occupational safety attitude and Gathers, quit intention . The correlations between occupational safety attitudes and occupational exposure were analyzed with Simple Linear Correlation.Results There were 94.30%female respondents,and only 5.70%male respondents.The average age of respondents was 22.52±2.90 years. The Simple Linear Correlation analysis showed that correlation between Negative attitude and occupational exposure was positive correlated, the occupational exposure and Support from managers, and Optimistic were negative correlation ( <0.05) .Conclusions Occupational safety and attitude are related to occupational exposure. Nursing students should apply standardized operation to improve the professional quality and ability. Schools should strengthen the education of occupational protection for nursing students. Hospital management should regularly organize nurses to study occupational exposure protection knowledge,give full affirmation of the nurse's achievements in the work, and offer certain reward appropriately.

Objective To explore the anti-inflammatory effect of antiflammin-2 (AF2) and recombinant peptide sequence 2(R2) on acute lung injury of mouse. To observe the expression of clara cell 16000 protein (CC16) and surfactant protein A (SP-A) in the lung of mouse inoculated with lipopolysaccharide (LPS) and the impact of AF2,R2,and glucocorticoids(hydrocortisone,HC) may have on the expression of the CC16 and SP-A in the lung of mice with acute lung injury. Methods Balb/c mice were inoculated with LPS (5 mg/kg) by intraperitoneal injection to set up ALI mice model. Mice weighed from 15 g to 16 g were grouped into control group, model group and treated groups respectively treated with AF2, R2 or HC. Mice in the control group were injected with physiological saline solution, while mice in the other four groups were inoculated with LPS to induce acute lung injury. Then animals in the treated groups were treated with AF2, R2 or HC each on a dose of 2 mg/kg also through intraperitoneal injection,while those of the control group and the model group, were given equivalent physiological saline solution as a placebo. The respiratory rate of all of these animals were recorded 6 hours after the injection. And at the time point of 12 hour,all the mice were sacrificed for a preparation of the whole lung tissue for the sake of a pathological investigation ,or for extractions of RNA for a semiquantitative analysis of the expression of CC16 and SP-A within the lungs. Results (1) An obvious attenuation of the respiratory rates of the three treated groups were observed when comparing with that of the mice in the model group without any anti-inflammatory treatment. (2) Remarkable extenuation of the extent of intra-alveolar and intersticial hemorrhage and infiltration of inflammatory cells were observed within the treated groups comparing with that of the model group. (3) An attenuate expressions of CC16 or SP-A were observed in the model group,while obvious uptrend of CC16 expression was observed in AF2 treated groups and increase of SP-A expressions were found in R2 and HC treated groups. Conclusion The anti-inflammatory effect of the peptide, AF-2 or R2, has been conformed on ALI mice model induced by LPS.

OBJECTIVETo investigate the effects of erlong zuoci (ELZC) pills and its disassembled prescriptions (Shudi-huang-Zexie group and Zexie group) on the enzymatic activity and protein expression changes of the key apoptosis molecules in the gentamycin injured hair cells.

METHODThe model of gentamycin induced ototoxicity in mice cochlear primary cultures was copied. Cochlear organotypic cultures of postnatal day 3-5 (P3-P5) mice were treated with gentamycin alone or in combination with ELZC pills, Shudihuang-Zexie group or Zexie group respectively. The enzymatic activity of Caspase-9 and Caspase-3 was determined by means of fluorescence staining in situ. The protein expression of Bcl-2 and Bax in the hair cell area was examined by immunofluorescence in normal and treated specimens.

RESULTAverage optical density analysis indicated that, compared to the normal group, 0.03 mmol x L(-1) gentamycin could significantly activate Caspase-9 and Caspase-3, downregulate the ratio of Bcl-2 and Bax protein expression. Compared to the gentamycin model group, ELZC pills significantly inhibited the enzymatic activity of Caspase-9 and upregulated the ratio of Bcl-2 and Bax protein expression, showing inhibition trend toward the enzymatic activity of Caspase-3. Both Shudihuang-Zexie group and Zexie group could effectively inhibit the enzymatic activity of Caspase-9 and Caspase-3, upregulate the ratio of Bcl-2 and Bax protein expression.

CONCLUSIONELZC pills, Shudihuang-Zexie group and Zexie group can effectively protect hair cells from gentamycin by correcting the abnormal changes of the mitochondrion-dependent signal transduction pathway.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Gentamicins ; adverse effects ; Hair Cells, Auditory ; cytology ; drug effects ; Male ; Mice ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism