[Diagnostic value of anti-Sa antibody and anti-carbamylated protein antibody for rheumatoid arthritis].

Objective: To investigate the diagnostic value of anti-Sa antibody and anti-carbonylated protein (CarP) antibody for rheumatoid arthritis (RA). Methods: A retrospective selection of 180 patients with RA who came to Qinghai Provincial People's Hospital from September 2022 to February 2023. Grouped according to the Disease Activity Score of 28 joints (DAS28), 101 of them were patients with RA in high activity (RAH group), 24 males and 77 females, aged 18-79 (53.2±12.2), and 79 patients with RA in low activity (RAL group), 23 males and 56 females, aged 24-78 (49.0±12.9).A total of 90 patients with other autoimmune diseases in the hospital in the same period were choosed as the other immune disease group, and 90 healthy physical examiners were as the healthy control group. The levels of serum anti-Sa and anti-CarP antibodies were measured by ELISA, RF by immunoscattering turbidimetry, anti-CCP by chemiluminescence, and ESR by Weil's method in four groups of patients. The area under the subject operating characteristic (ROC) curve (AUC) was applied to assess the sensitivity and specificity of each index alone or in combination for the diagnosis of RA. Results: In the RAH group, RAL group, other immune disease group, and healthy control group, the RF levels were 117.6 (61.0, 161.1), 92.7 (48.1, 92.7), 10.1 (5.3, 24.6), and 8.1 (6.0, 12.8) U/ml, anti-CCP antibody levels were 202.7 (67.1, 594.4), 212.9 (98.3, 416.2), 9.4 (6.6, 11.8), 1.9 (0.8, 4.9) U/ml, anti-Sa antibody levels were 305.3 (120.4, 614.9), 235.8 (161.6, 336.9), 123.9 (41.8, 240.5), 165.1 (71.1, 237.5) U/ml, and anti-CarP antibody levels were 11.7 (7.9, 21.6), 5.2 (3.3, 7.7), 5.1 (3.9, 6.5), and 5.8 (3.8, 7.5) mg/L, respectively, and their differences were statistically significant (all P<0.001). The level of anti-CarP antibody was higher in the RAH group than in the RAL group (P<0.001), and the difference in anti-Sa antibody was not statistically significant (P>0.05). The critical value of anti-Sa antibody at 181.45 µg/L showed a sensitivity of 67.2%, specificity of 65.6% and AUC of 0.710 (95%CI: 0.645-0.775); The sensitivity was 52.8% and the specificity was 88.9% with an AUC of 0.706 (95%CI: 0.646-0.766) at a critical value of 7.98 U/ml for the anti-CarP antibody. The AUC for the combined RF, anti-CCP antibody and anti-CarP antibody assay was 0.986 (95%CI: 0.977-0.996). Conclusion: Anti-CarP antibody is clinically significant in distinguishing active RA. RF, anti-CCP, and anti-CarP antibodies can be detected together with high AUC results, suggesting the potential for developing an improved method for diagnosing RA.

[Correlation of extracellular enzymes activity of Candida glabrata clinical isolates with in vivo pathogenicity in Galleria mellonella larvae].

Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC50) was calculated by using 1.25×108 CFU/ml,2.50×108 CFU/ml,3.75×108 CFU/ml,5.00×108 CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC50 concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC50=2.5×108 CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.

[Epidemiologic characteristics and drug resistance of isolated from blood culture escherichia coli in a hospital in Qinghai Province from 2016 to 2022].

Objective: To explore the drug resistance of Isolated From Blood Culture Escherichia coli (E. coli) in a hospital in Qinghai over the past seven years, to evaluate the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the homologous origin of E. coli, and to establish a protein fingerprint library to match with it, adjuvant clinical experience medication so as to provide the basis for the prevention and control of hospital-acquired infections. Methods: Retrospective analysis of blood cultures sent to hospitals from January 2016 to December 2022. Drug resistance and resistance changes in E. coli.A total of 1 841 E. coli strains were isolated from Qinghai Provincial People's Hospital from January 2016 to December 2022; all strains were identified by MALDI-TOF MS, and the VITEK2.0 drug sensitivity analyzer was applied for drug sensitivity analysis of the strains, and the mass spectrometry homology analysis and self-constructed protein fingerprint library were carried out by MALDI-Biotyper software; the protein fingerprint library was built by using WHONET5.6 software was used to statistically analyze the drug sensitivity results, SPSS23.0 software was used to analyze the relationship between fingerprint typing and drug sensitivity, and the χ2 test was used for intergroup comparisons. Results: A total of 1 841 strains of E. coli were detected in 4 582 positive blood culture specimens from January 2016 to December 2022, with a detection rate of 40.17%; the resistance rate of E. coli from blood sources to piperacillin/tazobactam and ceftriaxone was on the rise, and it was slightly decreased to cefepime, amikacin, levofloxacin, and sulfamethoxazole, and there was not much change to the rest of the drugs; After MALDI-Biotyper clustering analysis, the 1841 E. coli strains from Isolated From Blood Culture were classified into two major clusters and five subtypes, of which type Ⅰa1 accounted for about 40%, type Ⅰa2 accounted for about 2.7%, type Ⅰb accounted for about 3.8, type Ⅱa accounted for about 46%, and type Ⅱb accounted for about 7.5%. The detection rate of type Ⅰa1 E. coli was higher in general surgery (50.45%) and emergency surgery (50.92%), and the detection rate of type Ⅰb E. coli was higher in emergency medicine(10.05%)than in other departments. The drug sensitivity results of different subtypes were compared with each other, the resistance rate of type Ⅰa1 E. coli to cefepime was 21.3% higher than that of the remaining four types, and the difference was statistically significant (χ2=37.74,P=0.000); the resistance rate of type Ⅱ E. coli(>60%) to sulfamethoxazole was higher than that of type Ⅰ (<60%) as a whole, and the difference was statistically significant (χ2=15.248,P=0.004); and a preliminary database of homologous protein fingerprints of E. coli has been established E. coli homologous protein fingerprint library and validated. The drug susceptibility results of 1 288 E. coli strains in the validation set were statistically analyzed and compared with those in the training set. There was no significant difference(P>0.05). Conclusion: In recent years, the resistance rate of E. coli isolated from a hospital in Qinghai province to piperacillin/Tazobactam, cefepime, amicacin and other antibiotics has changed greatly. A fingerprint database of E. coli homologous protein was established, and it was found that the drug sensitivity data of E. coli were different among different fingerprint types. According to drug sensitivity, drug use could assist clinical experience and provide evidence for prevention and control of hospital illness.

[Application value of plasma proteomics in the diagnosis and pathogenesis of high altitude polycythemia].

Objective: To explore the application value of plasma proteomics in the diagnosis and pathogenesis of high altitude polycythemia (HAPC). Methods: Ten patients with HAPC in Qinghai Provincial People's Hospital from January 2020 to January 2021 were selected as the experimental group, including 4 males and 6 females, aged (46±4) years. Ten healthy controls at the same altitude in the same period were selected as the control group, including 5 males and 5 females, aged (44±4) years. The differential proteins were identified and quantified by high performance liquid chromatography-mass spectrometry (HPLC-MS), and the gene ontology (GO) functional enrichment analysis, the Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis and interaction network analysis were conducted for the selected differential proteins. Results: A total of 117 differential proteins with quantitative values were screened from the experimental group and the control group, 45 significantly up-regulated proteins with quantitative values only in the experimental group, 40 significantly down-regulated proteins with quantitative values only in the control group, and 32 differentially expressed proteins with quantitative values in both experimental and control groups were detected. Compared with the control group, 11 of the 32 differentially expressed proteins in the experimental group were down-regulated and 21 were up-regulated. The results of GO functional enrichment analysis showed that the biological processes involved by differential proteins mainly included immune response, complement activation, activated protein cascade and coagulation system. The results of KEGG function enrichment analysis showed that the main biochemical metabolic pathways and signal transduction pathways involved by differential proteins were axon guidance, lysosomes, cell adhesion molecules, lipid and atherosclerosis, hematopoietic cell lineage and cholesterol metabolism. The abundant domains are mainly in immunoglobulin-like domain, EGF-like domain, fibronectin type Ⅲ superfamily, serine proteases, Sushi/SCR/CCP superfamily. The results of differential protein interaction analysis showed that the interaction score was>700, and the top 10 differential proteins with the largest number of nodes were MPO, RPS27A, ARG1, GM2A, TIMP1, CRP, FABP5, HBB, S100A7 and RHOA, respectively. Conclusion: Plasma proteomics analysis technique is helpful to identify the related protein markers in the development of HAPC, and provide reference for the diagnosis and pathogenesis of HAPC.

[Application of next-generation sequencing technology detecting ctDNA in the diagnosis and treatment of gastric cancer].

Gastric cancer is a malignant tumor with high morbidity and mortality, which is a serious threat to human life and health. At present, the diagnosis and course monitoring of gastric cancer mainly rely on gastroscopic biopsy and CT, but their invasiveness and delay limit their clinical value. Gastric cancer patients urgently need a real-time, accurate, non-invasive diagnosis and course monitoring method. Circulating tumor DNA(ctDNA) is single or double stranded DNA released into body fluid by tumor cells, which contains the same genetic mutations as the original tumor cells. It is also one of the newly emerged biological markers. With the emergence and development of next-generation sequencing(NGS) technology, the sensitivity and specificity of ctDNA detection continue to increase, and it is gradually becoming a new choice for tumor diagnosis and disease course monitoring. In this paper, the current application of the next-generation sequencing technology detecting ctDNA in diagnosis and treatment of gastric cancer was discussed.

[Changes and clinical significance of serum NSE and MBP levels in children with cerebral palsy at high altitude during comprehensive rehabilitation].

Objective: To explore the changes of serum neuron-specific enolase (NSE) and myelin basic protein (MBP) in children with cerebral palsy at high altitude during comprehensive rehabilitation and their clinical significance. Methods: A clinical randomized controlled study design was used to select 144 children with cerebral palsy who were diagnosed and treated in the Rehabilitation Center of Xining Traditional Chinese Medicine Hospital of Qinghai Province from June 2018 to October 2019, including 83 males and 61 females, aged 3-5 years old. According to the order of admission, the random number table was used to divide into a conventional treatment group (n=72, 40 males and 32 females) and a comprehensive treatment group (n=72, 43 males and 29 females). The conventional treatment group was treated with conventional rehabilitation. The comprehensive treatment group was treated with monosialotetrahexose ganglioside sodium on the basis of conventional rehabilitation. In addition, 30 healthy children aged 3-5 years, 16 males and 14 females, were selected as the control group during the physical examination of the Pediatrics Department of Xining Hospital of Traditional Chinese Medicine, Qinghai Province. The serum levels of NSE and MBP in each group were detected, and the children's GMFM-88 scores were evaluated before and after treatment. The SPSS19.0 software was used for statistical analysis, the count data was tested by χ2. Results: The serum NSE and MBP levels of the control group were (5.96±0.80), (0.71±0.15) µg/L. Before treatment, the serum NSE and MBP levels of children with severe, moderate, and mild cerebral palsy were [(21.63±1.92), (3.63±0.49) µg/L], [(17.86±1.43) µg/L, (2.21±0.07) µg/L] and [(15.14±0.95), (1.76±0.30) µg/L], respectively. After treatment, the serum NSE and MBP levels of the conventional treatment group and the comprehensive treatment group were [(13.54±2.41), (2.07±0.85) µg/L] and [(12.09±2.37), (1.81±0.69) µg/L], respectively, and the GMFM-88 score was (116.75±27.41) points and (125.94±24.93) points. The levels of NSE and MBP in the serum of children with cerebral palsy were significantly higher than those of normal children in the control group, and their levels increased with the degree of disease, and the corresponding gross motor function scores were lower. After treatment, the GMFM-88 scale assessment scores of the two groups of children were significantly improved (t values were 310.97 and 70.86, P values were both<0.05), and serum NSE and MBP levels decreased to varying degrees compared with before treatment. The decline in the comprehensive treatment group was greater than that in the conventional treatment group. Conclusions: Serum NSE and MBP levels in children with cerebral palsy at high altitude are significantly higher than those in healthy children, and their levels are closely related to the degree of impairment and GMFM-88 scores in children with cerebral palsy. Dynamic monitoring of changes in NSE and MBP levels may be responsible for the condition and treatment effects of children with cerebral palsy judgments based.

Using modified fish scale waste from Sardinella brasiliensis as a low-cost adsorbent to remove dyes from textile effluents.

Textile industry wastewater has become a cause of concern to environmentalists due to its toxic composition and the difficulty of breaking down certain dyes. In this study, modified fish scales of Sardinella brasiliensis (SSb) were used as an alternative for a low-cost adsorbent to remove dyes from textile wastewaters. Adsorption efficiency was assessed by measuring the general, kinetic, and thermodynamic physico-chemical parameters of adsorption isotherms, using Reactive Turquoise Blue 15 (RTB15) and Reactive Red 120 (RR120) dyes as adsorbate models, as local textile industries commonly use these dyes. The isothermal data from the batch experiments were inserted in the Langmuir, Freundlich, and Langmuir-Freundlich (SIPS) equations; the Langmuir isotherm equation showed the most appropriate. The thermodynamic parameters showed that adsorption of dyes by the modified SSb adsorbent was an endothermic yet spontaneous process in the case of RR120. Sorbent-based on SSb material was concluded as adsorbing both of the tested dyes. Because of its abundant availability, and the small amount of activation needed to turn it into an adsorbent, this biowaste can be employed as a low-cost alternative for removal of dyes in the treatment of textile wastewater.

Natural impacted freshwaters: in situ use of alginate immobilized algae to the assessment of algal response.

The objective of this study was to investigate the feasibility of an in situ phytotoxicity test using alginate-immobilized algae for 60 days, in the assessment of water quality in an impacted small peri-urban stream. After laboratory optimization of algae immobilization/de-immobilization processes, the performance of immobilized/de-immobilized algae was compared to the performance of free algae in terms of specific algal growth and sensitivity. This was done by comparing 72 h EC50 values obtained with zinc and the pesticides clomazone and carbofuran. The results showed a similar performance, which allow us to conclude that immobilization for 60 days do not cause any significant alteration in algae physiology. In the field, immobilized algae were exposed at different times (2, 4 and 7 days) to water samples in both disturbed and undisturbed sites. Both laboratory and field experiments indicated that alginate-immobilized algae for 60 days were sufficiently sensitive for use in the in situ assessment of water quality.

Remediation of phenol-contaminated soil by a bacterial consortium and Acinetobacter calcoaceticus isolated from an industrial wastewater treatment plant.

Time-course performance of a phenol-degrading indigenous bacterial consortium, and of Acinetobacter calcoaceticus var. anitratus, isolated from an industrial coal wastewater treatment plant was evaluated. This bacterial consortium was able to survive in the presence of phenol concentrations as high as 1200mgL(-1) and the consortium was more fast in degrading phenol than a pure culture of the A. calcoaceticus strain. In a batch system, 86% of phenol biodegradation occurred in around 30h at pH 6.0, while at pH 3.0, 95.2% of phenol biodegradation occurred in 8h. A high phenol biodegradation (above 95%) by the mixed culture in a bioreactor was obtained in both continuous and batch systems, but when test was carried out in coke gasification wastewater, no biodegradation was observed after 10 days at pH 9-11 for both pure strain or the isolated consortium. An activated sludge with the same bacterial consortium characterized above was mixed with a textile sludge-contaminated soil with a phenol concentration of 19.48mgkg(-1). After 20 days of bioaugmentation, the remanescent phenol concentration of the sludge-soil matrix was 1.13mgkg(-1).

A simple methodology to evaluate influence of H2O2 and Fe(2+) concentrations on the mineralization and biodegradability of organic compounds in water and soil contaminated with crude petroleum.

Simple measurements of H2O2 concentration or CO2 evolution were used to evaluate the effectiveness of the use of Fenton's reagent to mineralize organic compounds in water and soil contaminated by crude petroleum. This methodology is suitable for application in small treatment and remediation facilities. Reagent concentrations of H2O2 and Fe(2+) were found to influence the reaction time and temperature, as well as the degree of mineralization and biodegradability of the sample contaminants. Some H2O2/Fe(2+) combinations (H2O2 greater than 10% and Fe(2+) greater than 50mM) resulted in a strong exothermic reaction, which causes peroxide degradation and violent gas liberation. Up to 75% TOC removal efficiency was attained in water and 70% in soil when high H2O2 (20%) and low Fe(2+) (1mM) concentrations were used. Besides increasing the degree of mineralization, the Fenton's reaction enhances the biodegradability of petroleum compounds (BOD5/COD ratios) by a factor of up to 3.8 for contaminated samples of both water and soil. Our experiments showed that low reagent concentrations (1% H2O2 and 1mM Fe(2+)) were sufficient to start the degradation process, which could be continued using microorganisms. This leads to a decrease in reagent costs in the treatment of petroleum-contaminated water and soil samples. The simple measurements of H2O2 concentration or CO2 evolution were effective to evaluate the Fenton's reaction efficiency.

Proposal of a sequential treatment methodology for the safe reuse of oil sludge-contaminated soil.

In this study sequential steps were used to treat and immobilize oil constituents of an oil sludge-contaminated soil. Initially, the contaminated soil was oxidized by a Fenton type reaction (13 wt% for H(2)O(2); 10mM for Fe(2+)). The oxidative treatment period of 80 h was carried out under three different pH conditions: 20 h at pH 6.5, 20 h at pH 4.5, and 40 h at pH 3.0. The oxidized contaminated sample (3 kg) was stabilized and solidified for 2h with clay (1 kg) and lime (2 kg). Finally, this mixture was solidified by sand (2 kg) and Portland cement (4 kg). In order to evaluate the efficiency of different processes to treat and immobilize oil contaminants of the oil sludge-contaminated soil, leachability and solubility tests were performed and extracts were analyzed according to the current Brazilian waste regulations. Results showed that the Fenton oxidative process was partially efficient in degrading the oil contaminants in the soil, since residual concentrations were found for the PAH and BTEX compounds. Leachability tests showed that clay-lime stabilization/solidification followed by Portland cement stabilization/solidification was efficient in immobilizing the recalcitrant and hazardous constituents of the contaminated soil. These two steps stabilization/solidification processes are necessary to enhance environmental protection (minimal leachability) and to render final product economically profitable. The treated waste is safe enough to be used on environmental applications, like roadbeds blocks.