Objective To investigate the stabilization of betaine on the enzyme activity of ALT in human serum at different temperatures, and to evaluate the matrix effects of human serum with betaine among different analytical systems. Methods The enzyme activity of ALT in human serums with l mol/L betaine stored at 37 ℃, 25 ℃, 4 ℃ and - 20 ℃ respectively were measured. The matrix effect experiment was carried out according to the procedures specified in Clinical and Laboratory Standards Institute (CLSI)document EP14-A. Four analytical systems were the candidates for evaluation, and Roche Diagnostics (Roche)-Hitachi analytical system served as the reference system. The enzyme activity of ALT of 40 patient specimens and 3-level specimens ( high, medium and low) with 1 mol/L betaine were measured using the evaluated methods and the reference method respectively. The linear regression analysis was performed to compare the mean y value of 3-level specimens with the statistically defined limits ( the two-tailed 95% prediction interval) derived from 40 patient specimen data. Results The enzyme activity of ALT could be stabilized with betaine in serum, which remained to be 99% for 48 h at 37 ℃, 98% for4 d at 25 ℃, 97%for 4 weeks at 4 ℃, and 99% for 4 months at - 20 ℃. The enzyme activity of control serum decreased to 42%, 86%, 71%, 79% respectively. The mean y values of three specimens with the betaine were within the 95% prediction interval of the estimated mean y values, which suggested no obvious matrix effects in serum. Conclusion Betaine can be used in the development of enzyme ALT calibrator as its stabilizer.

Objective To solve the problem that the rat retina paraffin sections are easily exfoliated from the slides and each layer are easily separation and fracture,we need to find a way to improve the retina paraffin section method and evaluate the tissue fixation. Methods We used 4 fixation liquids including 10%paraformaldehyde,4%paraformaldehyde, 4% paraformaldehyde and 95% alcohol and glacial acetic acid mixed liquid (FAA fixatiue solution ) combined with paraformaldehyde to fix the retinal tissue, and observed the fixation efficacy under microscope after HE staining. Results The effects of 10%paraformaldehyde and 4%paraformaldehyde fixed samples showed moderate separation and fracture of retina,but the HE staining retinal slices pre-treated by the FAA fixafive solution had bright and uniform color,although occasionally some parts of the retina were exfoliated from the slide, but it was not easy to take off,and had complete structure without separation and rupture. Conclusion The retina paraffin section fixed by FAA ixafive solution with 4% paraformaldehyde is superior to pure paraformaldehyde, and the paraformaldehyde concentration has no obviously influence on HE staining results.

Objective:To study the compatibility and stability of coenzyme A for injection, adenosine disodium triphosphate and inosine injection. Methods:By simulating the clinical medication, the three drugs and 5% glucose injection were mixed together. The contents and relative substances of coenzyme A, adenosine disodium triphosphate and inosine were measured by HPLC. The changes in appearance, pH and insoluble particles were observed or tested at ambient temperature. Results:The mixed solution showed no signifi-cant changes in appearance, pH, number of insoluble particles, contents and relative substances of coenzyme A, adenosine disodium triphosphate and inosine in 4 h, while the mixed solution became turbid and the pH, number of insoluble particles and contents of the three drugs showed significant changes after 24-h storage. Conclusion:The mixed solution of coenzyme A for injection, adenosine dis-odium triphosphate and inosine injection in 5% glucose injection should be used up in 4 h at ambient temperature.

Objective: To determine total phenylethanoid glycoside and acteoside in Plantago Herba to provide reference for evaluating the quality of medicinal materials.Methods: With acteoside as the control sample, a UV visible spectrophotometric method was used to determine total phenylethanoid glycosides in Plantago Herba.An HPLC method was applied to determine acteoside in Plantago Herba , and the conditions were as follows: an ODS2 C 18 (150 mm× 4.6 mm ,5 μm) chromatographic column was used with acetonitrile-0.1% formic acid (13∶87) as the mobile phase at a flow rate of 1.0 ml·min-1 , the detection wavelength was 332nm, the column temperature was 30℃, and the sample volume was 10 μl.Results: The reference solution and the sample solution had the maximum absorption at 332 nm, and the linear relationship was good within the range of 0.003 1-0.155 0 mg·ml-1 (r=0.999 5).The content of total benzene alcohol glycosides in 3 batches of samples was 2.73% , 2.61% and 2.84% , respectively;acteoside over the range of 0.000 6-0.155 0 mg·ml-1 (r=0.999 1) showed a good linear relationship with peak area,the sample recovery was 98.5% and the RSD was 1.6% (n =6), and the acteoside content in 3 batches of samples respectively was 0.54% , 0.51% and 0.56%.Conclusion: The method is simple, accurate and reproducible, and can be used for the determination of total phenylethanoid glycosides and acteoside in Plantago Herba.

Objective To establish a method for determination of phenylethanoid glycoside and acteoside in Plantago Herba. Methods UV-visible spectrophotometric method was used for the determination of the content of phenylethanoid glycosides compounds in Plantago Herba. HPLC method was used for the determination of acteoside in Plantago Herba. Chromatographic column with C18 ODS2 (4.6 mm × 150 mm, 5 μm) was used. Acetonitrile-0.1%formic acid (13:87) was as mobile phase; the flow rate was 1 mL/min; the detection wavelength was 332 nm; the column temperature was 30 ℃; the sample volume was 10 μL. Results The contents of phenylethanoid glycoside in Plantago Herba from different producing areas were among 1.03%–3.47%. Acteoside with peak area over the 0.0062–1.55 mg range showed a good linear relationship; the sample recovery rate was 98.9%, and the RSD was 1.6%. The contents of acteoside in Plantago Herba from different producing areas was among 0.18%–0.56%. Conclusion The method is simple, stable and reproducible, which can be used for the determination of phenylethanoid glycoside and acteoside in Plantago Herba from different producing areas and provide experimental basis for quality control of Plantago Herba.

Objective Toinvestigatetheoperativeeffectandsafetyofendovascularstentingfor thetreatmentofsymptomaticvertebralarteryostialstenosis.Methods Fortypatientswithsymptomatic vertebral artery ostial stenosis and stenosis rate ≥70% were admitted to the Department of Neurology, China-Japan Friendship Hospital from November 2010 to January 2013 were enrolled retrospectively. All patients received endovascular stenting therapy,15 of them were implanted bare metal stents,and 25 were implanted drug eluting stents. The technical successful rate of operation,perioperation complications,and symptom remission rate of the patients were analyzed. At the same time,stroke and death incident as well as the related ischemic symptoms of the stent vascular feeding area in the follow-up period (13 to 36 months)wereobservedandtherestenosisratewasdocumented.Results Atotalof42stentswereimplanted in 40 patients,and the technical success rate was 100. 0%. The preoperative stenosis rate of vertebral artery ostial stenosis was 75% to 99%(mean 85 ± 7%);the postoperative stenosis rate was 0% to 20%(mean 6 ± 4%). There was no perioperative complication. The clinical symptoms of 19 patients disappeared completely,16 were improved significantly within the follow-up period,and the symptom remission rate was 87. 5%. No stent vascular feeding area related stroke and death occurred. Four patients had transient ischemic attack in posterior circulation,13 had restenosis after procedure (10 of them with bare mental stents and 3 with drug eluting stents). There was significant difference in restenosis rate between the bare mental stents andthedrugelutingstents(10/15vs3/25,P=0.001).Conclusion Endovascularstentingforthe treatment of the severe symptomatic vertebral artery ostial stenosis is a safe and efficient method. Although its restenosis rate is high,but it can improve the symptom of posterior circulation ischemia effectively.

Objective:To study the effects of broad-spectrum antibiotics and induced antibiotic-resistant bacteria on the efficacy of 5-fluorouracil (5-FU) chemotherapy for mice with colon cancer and to investigate the underlying mechanisms associated with anti-tumor immune responses.Methods:BALB/c mice were subcutaneously injected with CT26 colon cancer cells and randomized into four groups: tumor-bearing control group, antibiotic group treated with ampicillin, streptomycin and colistin, 5-FU group and anitibiotic+ 5-FU group. Tumor volumes and body weights were measured and recorded. Seven days after the last 5-FU treatment, the percentages of splenic immune cell subpopulations and proliferated CD8 + T cells after co-culturing with CT26 were analyzed by flow cytometry. Gut microbiota composition was detected by 16S rRNA sequencing and the bacteria in mesenteric lymph nodes (mLN) were isolated and cultured. Bone marrow-derive macrophages were stimulated with identified bacteria and the expression of M1 and M2 polarization markers were assessed by quantitative PCR. The proliferation of CD8 + T cells co-cultured with bacteria-treated macrophages was analyzed by flow cytometry. In addition, tumor-bearing mice were treated with 5-FU and oral gavage of bacteria isolated from antibiotic+ 5-FU group or PBS. Tumor volumes, gut microbiota composition and the percentages of proliferated CD8 + T cells co-cultured with CT26 were assessed. Results:Tumor volumes were larger and body weights were lower in the antibiotic+ 5-FU group than in the 5-FU group. The percentages of CD4 + T cells, CD8 + T cells and neutrophils did not varied significantly after using antibiotics, however, the percentage of monocytes was increased in the antibiotic group. The percentage of proliferated tumor-specific CD8 + T cells in the antibiotic+ 5-FU group was decreased compared with that in the 5-FU group. Compared with the control group and 5-FU group, antibiotic usage was associated with the changes in gut microbiota composition with decreased α diversity indexes. Escherichia coli, Klebsiella pneumonia, and Proteus mirabilis were isolated from mLNs of the antibiotic group, 5-FU group and antibiotic+ 5-FU group, respectively. Bone marrow-derived macrophages stimulated with Proteus mirabilis expressed arginase at a high level, which was a M2 polarization marker of macrophage, and associated with the decreased percentage of proliferated CD8 + T cells after co-culturing. Bacteria of the genus Proteus were enriched in the gut microbiota of 5-FU-treated tumor-bearing mice with the oral gavage of Proteus mirabilis. Although no significant inhibitory effect on tumor growth was observed, the oral gavage of Proteus mirabilis was associated with the decreased percentage of proliferated tumor-specific CD8 + T cells in vitro. Conclusions:Broad-spectrum antibiotics inhibited the efficacy of chemotherapy and the proliferation of tumor-specific CD8 + T cells, in which antibiotic-resistant bacteria might be involved.

Objective To construct a structural equation model to analyze the effect of various factors on the quality of life in stroke patients.Methods In this cross-sectional study,a total of 657 stroke patients from 2 ter-tiary A hospitals in Shanghai were enrolled as investigation subjects via convenience sampling methodology.General information questionnaire,Sense of Coherence Scale-13,Perceived Social Support Scale,Medical Coping Modes Questionnaire,National Institutes of Health Stroke Scale,Self-Esteem Scale and Stroke-Specific Quality of Life were utilized as the investigating tools;Mplus 7.4 was used to construct a structural equation model.Results The aver age score of quality of life in stroke patients was(178.30±30.30);neurological status,self-esteem were negatively cor-related with quality of life(P＜0.05);sense of coherence,social support,coping style were positively correlated with quality of life(P＜0.05);sense of coherence,taking a confrontational approach for coping,yielding and neurological status had a direct impact on quality of life(path coefficients were 0.471,0.169、0.135,-0.089,P＜0.05);social sup-port,self-esteem,sense of coherence and neurological status had an indirect impact on quality of life(path coefficients were 0.232,0.108,0.058,-0.058,P＜0.001).Conclusion The quality of life in stroke patients is influenced by multi-ple factors interactively,and sense of coherence is an important factor affecting the quality of life;adopting a confron-tational approach as coping styles,good social support and high self-esteem can improve the level of quality of life.

Objective To analyze the clinical characteristics of thyroid dysfunction induced by camrelizumab in the treatment of classical Hodgkin lymphoma(cHL)and explore the influencing factors.Methods The medical records of cHL patients treated with camrelizumab from January 1st,2017 to December 31st,2020,were collected.The clinical characteristics of thyroid dysfunction induced by camrelizumab were analyzed,and the influencing factors of adverse drug reactions(ADRs)were discussed.Results A total of 47 patients were included,12 patients(25.53%)experiencing thyroid dysfunction after receiv-ing camrelizumab.Among them,3 patients had hypothyroidism,7 had subclinical hypothyroidism,and 2 had subclinical hyperthy-roidism.The severity of ADRs was between grade 1 and 2(mild to moderate).None of the patients with thyroid dysfunction dis-continued camrelizumab.Thyroid dysfunction occurred between 1 and 22 months after camrelizumab treatment,with a median time of 6 months.2 patients with hypothyroidism treated with levothyroxine,the thyroid function returned to normal in 1 patient and was improved in the other.1 patient with subclinical hypothyroidism was treated with levothyroxine with the thyroid function continued to be abnormal.The rest of the 9 patients with thyroid dysfunction received no intervention,and 4 of them returned to normal,and others remained with no obvious change or loss in follow-up.Thyroid dysfunction was associated with baseline thyroid stimulating hormone level(P=0.03).The level of thyroglobulin antibodies and thyroid peroxidase antibodies was increased in 2 patients a-mong 3 patients with moderate thyroid dysfunction.Conclusion The incidence of thyroid dysfunction induced by camrelizum-ab in cHL was high.Camrelizumab-induced thyroid dysfunction was related to TSH baseline level(P=0.03).The severity was between grade 1 and 2,which was related to the level of TgAb and TPOAb.The subtype of thyroid dysfunction was mainly sub-clinical hypothyroidism with a long period.Patients could continue camrelizumab treatment,and the thyroid function of patients with grade 2 hypothyroidism was improved after the treatment with levothyroxine.

Objective: To establish a reference measurement procedure for the determination of human serum homocysteine by isotope dilution high performance liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS), and to apply it to establish sample target values for external quality assessment (EQA) in clinical laboratories. Methods: The reference method of Hcyquantification in our laboratory was establishedaccording to the method recommended by the Joint Committee for Traceability in Laboratory Medicine (JCTLM). The precision, trueness, specificity, residue and matrix effect of the method were evaluated. The reference method was applied to establish Hcy target values for samples of the second EQA in Shanghai of 2018. Results: The method detects 12.5μmol/L and 37.4μmol/L samples in three batches in three days, and the CV between batches is 1.03% and 2.10%, respectively. The measured values of Standard reference material (SRM) 1955 of National Institute of Standards and Technology (NIST) were within the specified uncertainty range. No matrix effect and carryover were observed. The second EQA data in 2018 showed that the average value of domestic reagent group was lower than that of reference method, and that of imported reagent group was higher than that of reference method. Conclusion Thereference measurement procedure of ID-LC/MS/MS was successfully established to determine the human serum homocysteine. It is expected to play a role in tracing the quantities of Hcy in clinical laboratories.