Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Objective:To investigate the diagnostic value of nucleic acid matrix-assisted laser desorption ionization-time of flight mass spectrometry（MALDI-TOF MS）in sputum smear-negative patients with nontuberculous Mycobacterial（NTM）pulmonary disease.Methods:Clinical data of 123 patients suspected of NTM pulmonary disease admitted in Public Health Clinical Center Affiliated to Shandong University between July 2022 and November 2023 were retrospectively analyzed. Bronchoalveolar lavage fluid（BALF）specimens were collected for MALDI-TOF MS assay and MGIT 960 culture. The diagnostic efficacy of MALDI-TOF MS for NTM pulmonary disease in patients with negative sputum smears for acid-fast bacilli was evaluated with receiver operating characteristic（ROC）curve. Statistical analysis was performed using SPSS 26.0 software and MedCalc statistical software.Results:Diagnosis of NTM pulmonary disease was finally confirmed in 66 out of the 123 suspected patients. It took 8 to 24 h for MALDI-TOF MS to identify NTM species and resistance. By MALDI-TOF MS，72 NTM strains were identified，with the Mycobacterium avium complex being the most prevalent（34 strains，47.22%），followed by the Mycobacterium abscessus complex（13 strains，18.06%）；resistance to macrolides was detected in 6 cases，while no resistance to aminoglycosides was found. It took 9 to 45 days for BALF MGIT 960 culture to identify NTM，and took 7 to 15 days for NTM typing and drug sensitivity testing. By BALF MGIT 960 culture，28 NTM strains were identified；and 1 case was found to be resistant to macrolides. Using confirmed diagnosis as the gold standard，MALDI-TOF MS demonstrated higher sensitivity，negative predictive value，and agreement rate compared to MGIT 960 culture（84.85% vs. 42.42%，81.13% vs. 56.32%，80.49% vs. 62.60%， χ2=25.667，8.998，9.664， P<0.05 or <0.01）. The area under ROC curve（AUC）for MALDI-TOF MS was significantly higher than that of MGIT 960 culture（0.801 vs. 0.642， Z=3.300， P=0.001）. Conclusion:Compared to MGIT 960 culture，MALDI-TOF MS exhibits superior diagnostic efficiency in detecting NTM pulmonary disease in patients with acid-fast bacilli smear-negative sputum，with advantage of rapidly identifying NTM species and resistance.

AIM: To observe the changes of corneal densitometry(CD)in patients with keratoconus after corneal cross-linking(CXL).METHODS: Retrospective study. A total of 32 patients(43 eyes)with keratoconus in Ningxia Eye Hospital from April 2020 to April 2022 were selected. Pentacam analysis system divided the cornea into three layers: anterior 120 μm, middle layer and posterior 60 μm, and divides it into five regions with diameters of 0-2, 2-6, 6-10, 10-12 mm and full diameter according to the diameter, and measures the CD in different ranges. The changes of CD were compared before operation and at 1, 3 and 6 mo after operation.RESULTS: There were differences in uncorrected visual acuity, best corrected visual acuity and intraocular pressure before and 6 mo after operation(all P<0.05), and there was no difference in corneal endothelial cells(P=0.477). CD reached its peak at 1 mo after operation, and decreased at 3 mo and 6 mo after operation, but it was still higher than that before operation. There is a significant positive correlation between CD and Kmax in the anterior layer and the whole layer(r=0.164, P=0.016; r=0.152, P=0.023).CONCLUSION: The values of CD peaked at 1 mo after CXL, then it gradually decreased, tending to become stable at 6 mo postoperatively.

The core components of the Hippo signaling pathway encompass upstream regulatory molecules,core kinase cascade complexes,and downstream transcriptional regulation complexes.This pathway modulates cellular behaviors by influencing the effector molecules of its core components and plays a pivotal role in immune regulation.Effector molecules,such as Yes-associated protein(YAP),transcriptional coactivator with PDZ-binding motif(TAZ),transcriptional enhanced associate domain transcriptional factor(TEAD),monopolar spindle-one binder(MOB1),large tumor suppressor(LATS),can stimulate fibroblast-like synovial cell migration and invasion in rheumatoid arthritis,regulate osteoarthritis disease progression,promote pathological new bone formation in ankylosing spondylitis,sustain submandibular gland development while delaying Sjogren's syndrome progression,mediate alpha-smooth muscle actin in systemic sclerosis,and refine the regulation of target genes associated with pulmonary fibrosis.This article provides an overview of the regulatory mechanisms involving Hippo signaling pathway-related effector molecules in the pathogenesis and progression of rheumatic immune system diseases,to serve as a reference for exploring novel therapeutic targets of rheumatic immune system diseases.

Objective To explore the effect of topical application of Fushan rheumatism external prescription on inflammatory cytokines and notch2 pathway in rats with collagen-induced arthritis(CIA).Methods 36 female Wistar rats were randomly divided into normal group(n=10)and model group(n=26).CIA model was successfully established in 20 rats,which were randomly divided into model group(n=10)and Fushan rheumatism external prescription group(n= 10).Treatment was initiated on day 14,and 0.4 mL ointment was evenly applied to the ankle joints of rats in Fushan rheumatism external prescription group.Normal group and model group were topical smeared with the same volume of normal saline.Activity was taken twice a day for 42 days.The joint swelling of rats was observed every week and the arthritis index was scored.Thereafter blood was collected from abdominal aorta,and then ankle joint,spleen,liver,and kidney of rats were taken out after the end of the interventions.The severity of arthritis and the pathological changes of ankle joint,liver and kidney were evaluated by HE staining;Inflammatory cytokines expression in serum were detected by ELISA;The expression of Notch2,Delta-like ligand protein 1(delta-like ligand protein-1,DLL1)and nuclear factor-κ Bp65(nuclear factor-κ Bp65,NF-κ Bp65)mRNA and protein in synovium of ankle joints and spleen were detected by qRT-PCR and Western blot,and its positive expression in ankle joints were detected by immunohistochemical method;The changes of liver and kidney function of rats in each group were detected in serum.Results Compared with normal group,the arthritis index score in model group were increased(P<0.01),joint injury and pathological score were increased(P<0.01),the levels of inflammatory cytokines TNF-α,IL-6,IL-17 and IFN-γ in serum were increased(P<0.01),the expression of Notch2,DLL1 and NF-κBp65 mRNA and protein in joints and spleen were increased(P<0.01),and the positive expression in joints were also increased(P<0.01);Compared with model group,the arthritis index score in Fushan rheumatism external prescription group were decreased(P<0.05,P<0.01),joint injury and pathological score were decreased(P<0.01),the levels of inflammatory cytokines TNF-α,IL-6,IL-17 and IFN-γ in serum were decreased(P<0.01),the expression of Notch2,DLL1 and NF-κBp65 mRNA and protein in joints and spleen were decreased(P<0.01),and the positive expression in joints were also decreased(P<0.01).In addition,no noticeable tissue damages were observed in liver and kidney in Fushan rheumatism external prescription group,and the levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),blood urea nitrogen(BUN)and creatinine(Cr)in serum were no changes(P>0.05).Conclusion Fushan rheumatism external prescription relieves arthritis symptoms and joint injury in CIA rats,and has no toxic and side effects on liver and kidney.Its mechanism may be related to the reduction of inflammatory cytokines and down-regulation of Nocth2 pathway.

OBJECTIVE To study the vasorelaxant effects and mechanism of polyphenol compound 2，4′，5′-trihydroxy-5，2′- dibromo-diphenyl-methanone（LM49）on isolated aortic rings of rats. METHODS Thoracic aortic vascular rings of rats were collected. Using the diastolic rate as index ， the effects of different concentrations of LM 49 on endothelium-intact and endothelium-denuded aortic rings pre-contracted by norepinephrine （NE，1×10－6 mol/L）or KCl （60 mmol/L）were investigated. After pre-culturing vascular rings by nitric oxide synthase inhibitor L-nitro-arginine methyl ester （L-NAME，0.1 mmol/L） and cyclooxygenase inhibitor indomethacin （1×10－5 mol/L），as well as pre-culturing vascular rings by 4 potassium channel blockers [BaCl 2（0.1 mmol/L），tetraethylammonium（TEA，5 mmol/L），4-aminopyridine（4-AP，0.1 mmol/L）and glibenclamide （1×10－5 mol/L）]，the vasorelaxant effect of different concentrations of LM 49 on the vascular rings were investigated by using the same method. With the percentage of vasoconstriction as the index ，using KCl （60 mmol/L），NE（1×10－6 mol/L），calcium channel blocker verapamil （1×10－6 mol/L）and sarcoplasmic Δ 基金项目 重大新药创制国家科技重大专项 （No.2018ZX097- reticulum Ca 2 +-adenosine triphosphate （ATP） enzyme pump inhibitor thacarotene （TG，1×10－6 mol/L）to induce the release of calcium in vascular rings in the absence of calcium. CaCl was added cumulatively ，and the effect of LM 49 on the cxyw06，vasoconstriction caused by calcium influx induced by CaCl 2 was investigated. RESULTS 3×10－6，5×10－6，1×10－5 mol/L LM49 had a significant relaxation effect on NE and KCl precontracted vascular rings （P＜0.01）； whether the endothelium was removed or not had no significant effect on the vasodilation of LM 49（P＞0.05）. After L-NAME ，indomethacin， TEA and 4-AP was pre-incubated ，different concentrations of LM 49 had no significant effects on aortic rings precontracted by NE （P＞0.05）. Glibenclamide and BaCl 2 could inhibit the vasorelaxant effects of LM 49 on aortic rings precontracted by NE （P＜0.01）. In the absence of calcium ，LM49 could inhibit the contraction caused by calcium influx induced by accumulated CaCl 2 after pre-incubation with KCl ，NE，verapamil and TG （P＜0.01）. CONCLUSIONS LM49 evokes significant relaxation of isolated aortic vascular rings without endothelium dependence ；the mechanism of which is inducing ATP-sensitive potassium channel ， inward rectifier potassium channel open and restraining extracellular Ca 2 + influx via voltage-gated calcium channel ， receptor-operated calcium channel and store-operated calcium channel.

Psoriatic arthritis is a chronic systemic autoimmune disease, characterized by psoriasis skin lesions and inflammation of the spine and joint. It has complicated clinical manifestations and individual variations. Nearly half of the patients will have joints erosion in two years, which is crippling. The severity of the skin and joint disease frequently do not correlate with each other. Currently, the understanding of the disease is insufficient in China with the lack of standardized diagnosis and treatment. Therefore, researchers from the Chinese Rheumatology Association formulated this specification based on the diagnosis and management experience together with guidelines at home and abroad. The specification summarizes the present situation of domestic diagnosis and treatment, aiming to standardize the diagnosis process and treatment protocols of psoriatic arthritis. Furthermore, it can reduce misdiagnosis and missed diagnosis, as well as improve the prognosis.

Objective:To investigate the clinical efficacy of different doses of budesonide/formoterol in the treatment of an acute exacerbation of chronic obstructive pulmonary disease in patients.Methods:A total of 200 inpatients with an acute exacerbation of chronic obstructive pulmonary disease graded C/D by global initiative for chronic obstructive lung disease (GOLD) staging system who received treatment in Jiuquan City People's Hospital, China from January to December in 2019 were included in this study. They were randomly divided into a control group and a treatment group ( n = 100/group). Based on anti-infection and expectorant treatment, the treatment group was given inhalation therapy (higher dose budesonide/formoterol, 320 μg/9 μg), while the control group was identically given inhalation therapy (lower dose budesonide/formoterol, 160 μg/4.5 μg), with a total course of 9 days in each group. Before treatment and at 5 and 9 days of treatment, procalcitonin (PCT), high-sensitivity C-reactive protein (hs-CRP), fractional exhaled nitric oxide (FeNO), percentage of eosinophils (EO%), partial pressure of oxygen (PaO 2), lactic acid, interleulin-6 (IL-6), forced expiratory volume in 1 second (FEV 1), the ratio of FEV 1/ forced vital capacity (FVC) were monitored in each group. COPD assessment test (CAT) score, modified Medical Research Council (mMRC) dyspnea score, 6-minute walking test (6MWT), and symptom improvement were determined in each group. Results:Before treatment, there were no significant differences in PCT and PaO 2 between the control and treatment groups (both P > 0.05). There were significant differences in PCT, PaO 2, FeNO, hs-CRP, E0 (%), IL-6, FEV 1, FEV 1/FVC, 6MWT, mMRC, cough, expectoration, shortness of breath and CAT score measured at 5 days of treatment between the treatment and control groups ( t = 2.416, 3.289, 3.982, 4.871, 3.332, 4.098, 5.253, 6.214, 3.843, 7.268, 5.387, 7.392, 5.398, 6.349, all P < 0.05). There were significant differences in PCT, PaO 2, FeNO, hs-CRP, E0 (%), IL-6, FEV 1/FVC, FEV 1, 6MWT, mMRC, cough, expectoration, shortness of breath and CAT score measured at 9 days of treatment between the treatment and control groups ( t = 2.508, 4.032, 2.948, 3.527, 3.118, 5.251, 5.325, 6.338, 2.907, 6.289, 3.246, 2.084, 2.151, 2.527, all P < 0.05). At 5 days of treatment, lactic acid level in the observation group was significantly lower than that in the control group ( t = 4.341, P < 0.05). At 9 days of treatment, there was no significant difference in lactic acid level between the control and observation groups ( t = 1.173, P > 0.05). There was no significant difference in the incidence of adverse reactions between the control and treatment groups [4%(4/100) vs. 5%(5/100), P > 0.05]. Conclusion:Inhalation of high doses of budesonide/formoterol can greatly improve pulmonary function, 6MWT performance, decrease mMRC and CAT scores, alleviate cough, expectoration, shortness of breath, and decrease serum levels of FeNO, hs-CRP, E0(%), IL-6 and other inflammatory factors. Inhalation of higher doses of budesonide/formoterol exhibits better efficacy in the treatment of an acute exacerbation of chronic obstructive pulmonary disease in patients than inhalation of lower doses of budesonide/formoterol.

Objective: To analyze the epidemiological characteristics of varicella epidemic situation at schools and kindergartens in Fengxian district of Shanghai during 2012-2018, and to provide evidence for developing control and prevention strategies. Methods: Data of varicella epidemic situation at schools and kindergartens and varicella cases were analyzed by descriptive methods. The rates were compared with chi-square test. Results: A total of 6 727 varicella cases were reported in Fengxian during 2012-2018, for an incidence of 85.34 per 100 000 population. Totally 66 varicella epidemic outbreaks and public health emergencies caused by varicella were reported, which involved 741 varicella cases. Among them, 426 were breakthrough cases (57.49%). The average interval time of breakthrough cases were (6.78±2.79)years, and the incidence was more concentrated in the 3-9 years after vaccination with varicella vaccine. Most breakthrough cases appeared 5 years past vaccination. The peak time of varicella was from April to June and from October to December. Most of cases were in the 5-11 year-old group and 15-17 year-old group. The cases in males was higher than in females. Kindergarten childcare and primary school children were at high risk of varicella. Timeliness of reporting was positively correlated with the duration of the varicella epidemic situation( r =0.52， P <0.05). Conclusion Most varicella epidemic situation in Fengxian district of Shanghai occued in primary schools during autumn and winter, and most of the varicella cases were breakthrough cases. Emergency vaccination of varicella vaccine can effectively control the spread of the epidemic. It is suggested to increase the publicity of the immunization program of 2 doses of varicella vaccine and raise the awareness of varicella vaccination of children’s parents.

From September 2013 to October 2018, 39 patients (28 males and 11 females, aged 21 to 76 years) with stage 4 pressure ulcers were admitted to the General Hospital of Xinjiang Military Command. The area of pressure ulcers ranged from 2 cm×2 cm to 20 cm×12 cm on admission. The two-stage method of debridement and skin flap transfer was exploited to repair the wounds. In the first stage, a thorough debridement was performed (26 cases underwent debridement once, 10 cases twice, and 3 cases for three times). The skin flap transfer surgery was conducted in the second stage after 6 to 12 days (local skin flap for 16 cases, vascularized island flap for 8 cases, fascial flap for 5 cases, gluteus maximus flap for 5 cases, and biceps femoris flap for 5 cases), with flap area of 4 cm×2 cm to 16 cm×10 cm. Some donor sites were closed by direct suture and the other donor sites which can not be sutured were covered by medium-thickness skin graft from the lateral thigh. All the pressure ulcers of 39 cases were healed with no sinus. During follow-up of 6 months to 5 years, no recurrence of pressure ulcer at the surgical site was observed; the flaps achieved soft texture and good appearance. Thus, the two-stage method of debridement and skin flap transfer achieved good long-term curative effect and could be a preferable option for treating stage 4 pressure ulcers.