There are 2 techniques for detecting red blood cell survival(RBCS)detection techniques:red blood cell labeling test and carbon monoxide(CO)breath test.The former has disadvantages such as long measurement times and complicated procedures,while the latter is simple,convenient,moderately priced,and capable of dynamically monitoring changes in RBCS before and after treatment.Currently,the CO breath test is gradually being implemented in clinical practice.RBCS is not only applied to hematologic diseases such as multiple myeloma,myelodysplastic syndromes,lymphoma,and thalassemia,but also to non-hematologic diseases like type 2 diabetes and chronic kidney disease.It can assist in diagnosis,guide treatment,evaluate drug treatment efficacy,and predict disease progression.

To evaluate the clinical value of targeted next generation sequencing (tNGS) in diagnosing rifampicin and rifabutin resistance in tuberculosis patients. In this retrospective cohort study, 119 culture-positive Mycobacterium tuberculosis (MTB) strains from tuberculosis patients in Shenzhen Third People′s Hospital from 2020 to 2023 were collected, then tNGS was performed to detect mutations of rpoB gene. Fourteen different types of rpoB gene mutation were detected in 46 mutation MTB strains, including 43 resistance related mutations and 3 synonymous mutations at codon 529. Using the phenotypic drug susceptibility results of rifampicin and rifabutin as the reference standard, the sensitivities of tNGS for detecting resistance to rifampicin and rifabutin were 100%, and the specificities were 96.2% and 89.4% respectively, therefore, tNGS showed good diagnostic performance. Mutations at positions 531 and 526 of rpoB were highly associated with resistance to rifampicin and rifabutin. Moreover, the results of tNGS from the clinical specimens were consistent with those from the corresponding culture strains. tNGS analysis was performed on 83 MTB strains from 18 patients with multiple positive cultures. The results showed that 2 patients with no mutations in the initial MTB strains were subsequently detected with rpoB gene mutation and their phenotypic drug susceptibilities changed from sensitive to resistant. In summary, using tNGS to detect rpoB mutations can reduce false positive results caused by synonymous mutations, and have satisfactory performance for the diagnosis of rifampicin and rifabutin resistance. tNGS can directly detect clinical sputum samples, and also can be used to dynamically monitor the molecular resistance profiles of MTB, therefore it has extremely broad clinical application prospects.

To evaluate the clinical value of targeted next generation sequencing (tNGS) in diagnosing rifampicin and rifabutin resistance in tuberculosis patients. In this retrospective cohort study, 119 culture-positive Mycobacterium tuberculosis (MTB) strains from tuberculosis patients in Shenzhen Third People′s Hospital from 2020 to 2023 were collected, then tNGS was performed to detect mutations of rpoB gene. Fourteen different types of rpoB gene mutation were detected in 46 mutation MTB strains, including 43 resistance related mutations and 3 synonymous mutations at codon 529. Using the phenotypic drug susceptibility results of rifampicin and rifabutin as the reference standard, the sensitivities of tNGS for detecting resistance to rifampicin and rifabutin were 100%, and the specificities were 96.2% and 89.4% respectively, therefore, tNGS showed good diagnostic performance. Mutations at positions 531 and 526 of rpoB were highly associated with resistance to rifampicin and rifabutin. Moreover, the results of tNGS from the clinical specimens were consistent with those from the corresponding culture strains. tNGS analysis was performed on 83 MTB strains from 18 patients with multiple positive cultures. The results showed that 2 patients with no mutations in the initial MTB strains were subsequently detected with rpoB gene mutation and their phenotypic drug susceptibilities changed from sensitive to resistant. In summary, using tNGS to detect rpoB mutations can reduce false positive results caused by synonymous mutations, and have satisfactory performance for the diagnosis of rifampicin and rifabutin resistance. tNGS can directly detect clinical sputum samples, and also can be used to dynamically monitor the molecular resistance profiles of MTB, therefore it has extremely broad clinical application prospects.

Objective: To explore the effect of salvianolate combined with the conventional therapy on acute ischemic stroke and observe the influence on serum inflammatory cytokines of interleukin-6(IL-6).Methods: According to the random number table, 70 patients were randomly divided into the observation group (n =35) and the control group (n =35), and both were given the conventional therapy for acute ischemic stroke.The observation group was given intravenous injection of 200mg salvianolate in 250ml normal saline once a day additionally.The treatment course was 2 weeks.Another 30 persons with physical examination were in the healthy control group.The neurologic damage deficiency score (NIHSS score) was evaluated after the 3-, 7-,10-and 14-day treatment in the groups, the serum IL-6 in 24 h after onset, and after the 3-, 7-,10-and 14-day treatment was detected and compared with that in the healthy control group.Results: After the 7-day treatment, NIHSS score in the observation group decreased significantly when compared with that on admission (P＜0.05), and remained the decreasing trend.After the 10-day treatment, NIHSS score in the control group decreased significantly when compared with that on admission (P＜0.05).After the 7-day treatment, NIHSS score in the observation group was lower than that in the control group (P＜0.05).Compared with that in the healthy control group, the serum level of IL-6 in the observation group and the control group was higher in 24 h of admission (P＜0.05).The serum level of IL-6 in the observation group decreased after the 7-day treatment, and was similar to that in the healthy control group after the 14-day treatment (P＞0.05).The serum level of IL-6 in the control group decreased after the 10-day treatment, while was higher than that in the healthy control group during the whole study period (P＜0.05).The serum level of IL-6 in the observation group was lower than that in the control group after the 7-day treatment (P＜0.05), and the peak value in the observation group was notably lower than that in the control group (P＜0.05).Conclusion: Salvianolate combined with the conventional therapy can effectively decrease the NIHSS score and the content of IL-6 in the patients with acute ischemic stroke, which shows better effect than the conventional therapy alone.

Objective To investigate the pharmacokinetics and relative bioavailability of praziquantel injection in buffaloes in contrast to praziquantel tablet. Methods A single oral administration of praziquantel tablet at a dose of 20 mg/kg or intramus-cular administration of praziquantel injection at a dose of 10 mg/kg was performed in six healthy adult buffalos according to a two-period crossover design. The praziquantel concentration in plasma was determined by a high performance liquid chromatography (HPLC)method. The pharmacokinetic parameters were calculated by non-compartmental analysis. Results The main pharma-cokinetic parameters of praziquantel tablet were as follows:Tmax=(0.60±0.29)h,Cmax=(0.57±0.37)μg/ml,T1/2β=(0.70±0.42) h,AUC=(0.80±0.70)(μg/ml)·h. The main pharmacokinetic parameters of praziquantel injection were as follows:Tmax=(0.65± 0.49)h,Cmax=(3.82 ± 1.17)μg/ml,T1/2β=(1.00 ± 0.73)h,AUC=(1.61 ± 0.89)(μg/ml)·h. The relative bioavailability of pra-ziquantel injection was 402.5％in contrast to praziquantel tablet. Conclusion The praziquantel injection has pharmacokinetic characteristics of rapid absorption,high bioavailability and extensive distribution,and the clinical recommended dosage of pra-ziquantel injection is 10 mg/kg.

Background:Acute-on-chronic liver failure( ACLF)is a commonly seen liver failure in China,and lacking an animal model that can effectively simulate the dynamic change of immune status of ACLF. Aims:To establish an animal model that can simulate dynamic change of immune status of ACLF by repeated administration of concanavalin A(ConA). Methods:Mice were randomly divided into normal control group and ConA repeated administration group. Mice in ConA repeated administration group were injected with ConA 15 mg/ kg through retrobulbar angular vein every 48 hours for 5 times,and mice in control group were injected with same volume of 0. 9% NaCl solution. Serum levels of IL-6,IL-10,IL- 12,TNF-α,IFN-γ,MCP-1 in peripheral blood were assessed by CBA assay,and the ratio of IL-10/ TNF-α was calculated. The expression of HLA-DR,number and proportion of CD4+ T cells and the expression of PD-1 of monocytes in peripheral blood were detected by flow cytometry. Results:Peripheral blood cytokines changed from predominated proinflammatory cytokines into predominated anti-inflammatory cytokines with the increasing in time of administration in ConA repeated administration group. Compared with control group,HLA-DR expression of monocytes in peripheral blood was significantly decreased(P <0. 05),number and proportion of CD4+ T cells were significantly decreased(P <0. 05), and PD-1 expression was significantly increased( P < 0. 05)in ConA repeated administration group. Conclusions:An animal model of ACLF immune status from systemic inflammatory response syndrome( SIRS) to compensatory antiinflammatory response syndrome(CARS)induced by repeated ConA stimulation is successfully established.

Objective To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by morphine preconditioning in the rats with chronic heart failure in vitro.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,aged 6-7 weeks,in which doxorubicin 2 mg/kg was injected via the tail vein once a week for 6 consecutive weeks to induce chronic heart failure,were studied.At the end of 8th week,30 rats with chronic heart failure were randomly divided into 5 groups (n =6 each) using a random number table:sham operation group (group Sham),I/R group,morphine preconditioning group (group MPC),SB203580 (p38MAPK inhibitor) + morphine preconditioning group (group SBM),and SB203580 group (group SB).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the anterior descending branch of the left coronary artery followed by 120 min of reperfusion to establish the model of myocardial I/R injury.After equilibration,the hearts were subjected to 3 cycles of 5 min perfusion with K-H solution containing morphine 1 μmol/L at 5-min intervals before ischemia in group MPC.In group SBM,the hearts were perfused with K-H solution containing SB203580 (5 μmol/L) for 45 min starting from l0 min before morphine preconditioning until 5 min of ischemia.In group SB,morphine preconditioning was not performed,and the hearts were only perfused with K-H solution containing SB203580 (5 μmol/L) starting from 40 min before ischemia until 5 min of ischemia.At 15 min of equilibration (baseline),5 and 10 min of reperfusion,the coronary effluent was collected to detect the activity of lactate dehydrogenase (LDH) using the chemical colorimetry.At 10 min of reperfusion,the expression of phosphor-p38MAPK (p-p38MAPK) in the myocardium was determined by Western blot in Sham,I/R and MPC groups.At 120 min of reperfusion,the area at risk (AAR),total areas of right and left ventricles (LV+RV),and infarct size (IS) were measured,and the IS/AAR ratio was calculated.Results Compared with group Sham,the LDH activity in coronary effluent during reperfusion and IS/AAR ratio were significantly increased in the other groups,and the expression of p-p38MAPK was significantly up-regulated in I/R and MPC groups (P＜0.05).Compared with group I/R,the LDH activity in coronary effluent during reperfusion was significantly decreased,the expression of p-p38MAPK was significantly up-regulated,and the IS and IS/AAR ratio were significantly decreased in group MPC (P＜0.05),and no significant change was found in the LDH activity in coronary effluent,IS and IS/AAR ratio in SBM and SB groups (P＞0.05).Compared with group MPC,the LDH activity in coronary effluent during reperfusion was significantly increased,and the IS and IS/AAR ratio were significantly increased in group SBM (P＜0.05).Conclusion The mechanism by which morphine preconditioning reduces myocardial I/R injury is related to activation of p38MAPK signaling pathway in the rats with chronic heart failure in vitro.

Aim To evaluate the effects of morphine preconditioning ( MPC ) on the expression of microR-NAs ( miRNAs ) induced by hypoxia-reoxygenation (H/R) in H9c2 myocardial cells. Methods H9c2 cells were randomly divided into 3 groups ( n=4 each) as follows:control group ( CON) , hypoxia/ reoxygen-ation group ( H/R ) and morphine preconditioning group ( MPC+H/R) . The cells were cultured in nor-mal condition in CON group. The cells were subjected to 5 h hypoxia followed by 1 h reoxygenation in H/R group and MPC+H/R group. Specifically, the cells in MPC+H/R group were preconditioned with morphine with the final concentration of 1 μmol·L-1 for 10 min before H/R. After the treatment, CCK-8 was used to detect cell viability and chemical colorimetry was used to detect lactate dehydrogenase ( LDH ) activity in the culture medium. Cell apoptosis was assessed by An-nexin-V-FITC/PI flow cytometry. Relative expression of Fas protein was detected by Western blot. The ex-pression of miRNA in myocardial cells was analyzed by quantitative reverse transcription polymerase chain re-action ( qRT-PCR ) . Results Compared with CON group, the cell viability was significantly decreased, while the LDH activity, apoptotic rate and Fas protein expression were dramatically increased in group H/R (P<0. 01). However, MPC significantly increased the cell viability, whereas it decreased the LDH activity, apoptotic rate and Fas protein expression induced by H/R injury ( P < 0. 01 ) . The expressions of miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7 e-5 p were markedly down-regulated by H/R as compared to CON group ( P <0. 05 ) , while MPC inhibited these miRNAs which were significantly down-regulated by H/R group ( P <0. 01 ) . Conclusion Morphine preconditioning might protect H9 c2 myocar-dial cells against H/R injury by regulating the expres-sion of miRNAs such as miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7e-5p.

Objective To compare the advantages and disadvantages of high intensity focused ultrasound (HIFU ) and total abdominal hysterectomy for the treatment of uterine fibroids .Methods 167 patients with symptomatic uterine fibroids were divid‐ed into the HIFU treatment group(n=86) and the hysterectomy group(n=81) .The adverse events were recorded before operation and at postoperative 1 ,3 ,6 months .The questionnaire survey of the uterine fibroid symptoms‐quality of life (UFS‐QOL) was con‐ducted .The health related living quality was evaluated by using the health survey questionnaire‐36(SF‐36) .Results There were no severe adverse events in either group .The significant clinical complications and adverse events in the HIFU group were lower than those in the hysterectomy group .The UFS‐QOL scores in the HIFU group were superior to those in the hysterectomy group ,but the SF‐36 scores at postopertive 3 ,6 months had statistically significant differences between the two groups (P>0 .05) .Conclusion The patients who are unable to tolerate surgery or hope to preserve the uterus and its physiological function are suitable for HI‐FU treatment .

Objective To discuss the test efficiency of three methods for detecting Toxoplasma IgG antibody. Methods To-tally 304 specimens were detected parallelly for Toxoplasma IgG antibody by using the gold marked method,indirect hemagglutina-tion test(IHA),and enzyme-linked immunosorbent assay(ELISA),and the sensitivity,specificity and Youden index of these methods were compared. Results The detection sensitivities of gold marked method,IHA,and ELISA for Toxoplasma IgG anti-body were 85.5%,89.8%and 91.9%respectively(χ2=4.12,P>0.05);the specificities were 92.4%,96.6%and 97.5%respec-tively(χ2=4.06,P>0.05). The detection efficiency and Youden index of ELISA were 94.1%and 0.89 respectively,being high-er than those of IHA and gold marked method. Conclusion The sensitivity and specificity of the ELISA method for Toxoplasma IgG antibody are higher,and in addition,it can be automated. Therefore,it is suitable for large-scale Toxoplasma IgG antibody screening.