Objective To develop a multiplex PCR assay for simultaneous detection of four intestinal parasites, including Giardia duodenalis, Cryptosporidium parvum, Enterocytozoon bieneusi and Moniezia, and to preliminarily evaluate its detection efficiency. Methods Four pairs of specific primers were designed based on the conserved sequences of the corresponding genes of G. duodenalis (GenBank accession number: XM_001710026.2), C. parvum (GenBank accession number: XM_626998.1), E. bieneusi (GenBank accession number: KJ719492.1) and Moniezia (GenBank accession number: OM296991.1) retrieved from the GenBank database, and a multiplex PCR assay for simultaneous detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia was developed and optimized. A total of 116 fresh goat stool samples were collected from four goat farms in Zhanjiang City, Guangdong Province during the period from October to December 2022, including 96 samples used for evaluating the detection efficacy of the multiplex PCR assay, and 20 samples as baseline controls for sample testing. Genomic DNA extracted from 96 goat stool samples was tested using the single-target PCR assay and the developed multiplex PCR assay, and the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex PCR assay were evaluated for detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia DNA in goat stool samples with the single-target PCR assay as the gold standard. Results The multiplex PCR assay developed in this study allowed simultaneous amplification of specific gene fragments of G. duodenalis, C. parvum, E. bieneusi and Moniezia, with 1 400, 755, 314 bp and 585 bp in sizes, respectively, and the detection limit was 10² and higher copies of parasite DNA clones, while the multiplex PCR assay was negative for gene amplification of Schistosoma japonicum, Fasciola hepatica, Echinococcus granulosus, Blastocystis hominis and Homalogaster paloniae. Single-target PCR assay and the developed multiplex PCR assay were employed to test DNA samples extracted from 96 goat stool samples, and single-target PCR assay tested positive in 40 goat stool samples (41.67%), including 39 positive samples tested with the multiplex PCR assay, with a mean coincidence rate of 97.50% (39/40). The multiplex PCR assay tested positive for G. duodenalis DNA in 26 goat stool samples (27.10%), C. parvum DNA in 22 samples (22.90%), E. bieneusi DNA in 24 samples (25.00%), and Moniezia in 9 samples (9.40%), which was consistent with the detection using the single-target PCR assay. The sensitivity, negative predictive value, and positive predictive value of the multiplex PCR assay were 96.15%, 95.83%, 100.00% and 100.00%, 98.90%, 98.92%, 100.00% and 100.00%, 100.00%, 100.00%, 100.00% and 100.00% for detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia DNA in goat stool samples, respectively, if the single-target PCR assay served as the gold standard. Conclusion A highly sensitive and specific multiplex PCR assay has been developed for simultaneous detection of G. duodenalis, C. parvum, E. bieneusi and Moniezia in goats, which is suitable for rapid, large-scale screening of intestinal parasites in sheep stool samples.

Bladder cancer remains the 10th most common cancer worldwide. In recent years, metformin has been found to have potential anti-bladder cancer activ-ity while high concentration of IC50 at millimolar level is needed, which could not be reached by regular oral administration route. Thus, higher efficient agent is urgently demanded for clinically treating bladder cancer. Here, by conjugating artesunate to metformin, a novel artesunate-metformin dimer triazine derivative AM2 was designed and synthesized. The inhibitory effect of AM2 on bladder cancer cell line T24 and the mechanism underlying was determined. Anti-tumor activity of AM2 was assessed by MTT, cloning formation and wound healing assays. Decreasing effect of AM2 on lipogenesis was determined by oil red O staining. The protein expressions of Clusterin, SREBP1 and FASN in T24 cells were evaluated by Western blotting. The results show that AM2 significantly inhibited cell proliferation and migration at micromolar level, much higher than parental metformin. AM2 reduced lipogenesis and down-regulated the expressions of Clusterin, SREBP1 and FASN. These results suggest that AM2 inhibits the growth of bladder cancer cells T24 by inhibiting cellular lipogenesis associated with the Clusterin/SREBP1/FASN signaling pathway.

Objective:To systematically review and evaluate the safety and efficacy of high-flow nasal oxygen (HFNO) for pre-oxygenation before anesthesia induction.Methods:Pubmed, Cochrane Library, Embase, China National Knowledge Infrastructure, Wanfang Database, China Science and Technology Journal Database and China Biomedical Literature Database were searched from inception to March 2022.All randomized controlled trials comparing HFNO and facemask ventilation for pre-oxygenation before anesthesia induction were collected.Two researchers independently assessed the quality of trials and extracted data.The primary outcome was the safe apnea time, the secondary outcomes were the lowest SpO 2 during intubation, oxygenation-related complications, patient comfort, PaO 2 and PaCO 2 before and after pre-oxygenation and after intubation.Meta-analysis was performed using RevMan 5.4 software. Results:Seventeen randomized controlled trials involving 843 patients were included in this meta-analysis.The results of meta-analysis showed that the safe apnea time was significantly longer ( MD=67.61, 95% CI 5.94-129.28, P=0.03), the lowest SpO 2 was higher during tracheal intubation ( MD=3.27, 95% CI 2.25-4.29, P<0.01), and PaO 2 was higher after pre-oxygenation ( MD=54.39, 95% CI 9.32-99.46, P=0.02) in the patients using HFNO than those using facemask ventilation.There were no statistically significant differences in the other outcomes ( P>0.05). Conclusions:HFNO for pre-oxygenation before anesthesia induction can significantly prolong the safe apnea time, increase the lowest SpO 2 during tracheal intubation, and improve the levels of PaO 2 after pre-oxygenation, and HFNO does not affect the patient′s comfort or increase the development of preoxygenation-related complications when compared with facemask ventilation.

Objective:To investigate the knowledge, attitudes, and practices (KAP) of pulse oximetry among pediatric healthcare providers in China and analyze the factor influencing the KAP.Methods:A self-developed questionnaire was used for an online research on the KAP of 11 849 pediatric healthcare providers from 31 provinces, autonomous regions, and municipalities of China from March 11 to 14, 2022.The factors influencing the KAP of pulse oximetry among pediatric healthcare providers were examined by Logistic regression. Results:The scores of KAP, of pulse oximetry were 5.57±0.96, 11.24±1.25 and 11.19±4.54, respectively.The corresponding scoring rates were 69.61%, 74.95%, and 55.99%, respectively. Logistic regression results showed that the gender and working years of pediatric healthcare providers, the region they were located, and whether their medical institution was equipped with pulse oximeters were the main factors affecting the knowledge score (all P<0.05). Main factors influencing the attitude score of pediatric healthcare providers included their knowledge score, gender, educational background, working years, region, medical institution level, and whether the medical institution was equipped with pulse oximeters (all P<0.05). For the practice score, the main influencing factors were the knowledge score, gender, age, and whether the medi-cal institution was equipped with pulse oximeters (all P<0.05). Conclusions:Chinese pediatric healthcare providers need to further improve their knowledge about and attitudes towards pulse oximetry.Pulse oximeters are evidently under-used.It is urgent to formulate policies or guidelines, strengthen education and training, improve knowledge and attitudes, equip more institutions with pulse oximeters, and popularize their application in medical institutions.

Objective To understand the distribution of allergens in children with common allergic diseases in Guangzhou, and to provide evidence for the prevention and treatment of environmental allergens in children with different allergic diseases. Methods This study enrolled a total of 6 869 children with allergic diseases, including wheezing diseases and asthma (1 013 cases), allergic rhinitis (660 cases), allergic conjunctivitis (422 cases), eczema (2 762 cases), atopic dermatitis (831 cases) and urticaria (1 181 cases). Western blotting was used to detect the serum allergen type E specific antibodies (sIgE) of the patients. The results were statistically analyzed. Results The total positive rate of sIgE in 6 869 children was 74.01%. The positive rate of male children (75.02%) was higher than that of female children (72.25%). The positive rate of inhalation allergens (53.47%) was higher than that of food allergens (57.39%). House dust mites and milk were the most common inhalation allergens and food allergens, respectively. Eczema was the most common allergic disease in infants and toddlers. Allergic rhinitis and urticaria were common in school-age children. Conclusion The main allergens causing allergic diseases in children in Guangzhou were dust mites and milk. Different allergic diseases had its specific allergens. It is necessary to take targeted prevention measures accordingly.

Objective:To make the early clinical antibiotic regimen by finding out the infection of corynebacterium in the pus of patients with granulomatous mastitis in the early stage.Methods:A total of 42 patients who were diagnosed and treated in the Breast Center of Guangdong Maternal and Child Health Hospital from Jun. 2016 to Mar. 2017 with complete follow-up data were retrospectively analyzed. PCR method was used to detect corynebacterium in the patients’ pus. Patients in the positive group were treated with antibiotics alone, antibiotics + hormones and hormones alone, while patients in the negative group were treated with antibiotics + hormones and hormones alone. The postoperative recurrence rate and cure rate of different groups of patients were observed.Results:The antibiotic regimen for granulomatous mastitis in patients with corynebacterium infection included a combination of short-acting levofloxacin and azithromycin and long-acting anti-mycobacterium drugs. Among the 42 patients in the subgroup, 21 patients were confirmed positive for corynebacterium by PCR detection of pus, and the postoperative recurrence rate was 23.5%. There was a statistically significant difference between the antibiotic group, the antibiotic + hormone group and the hormone group in treatment of granulomatous mastitis infected with corynebacterium ( χ2=5.494, P=0.036) . PCR detection shouwed corynebacterium negative in 21 cases, and postoperative recurrence rate of 16.7%. No statistically significant difference in efficacy was found between antibiotic + hormone group and hormone only group for GM patients without bacterial infection ( χ2=1.129, P=0.719) . Conclusion:Early detection of corynebacterium infection in GM patients is significant for clinical guidance of the application of lipophilic antibiotics.

Objective To investigate the clinical efficacy of one-stage posterior percutaneous transpedicular screw fixation combined with lateral anterior approach for focus debridement bone graft fusion in the treatment of lumbar tuberculosis(TB).Methods Eighty-seven cases of lumbar tuberculosis treated by operation respectively adopted the one-stage anterior and posterior combined approach operation(group A) and simple lateral anterior approach operation(group B).The levels of CRP and ESR,Cobb angle,ODI score and Frankl score before and after surgery were statistically analyzed,and operation situation was analyzed in the two groups.Results The operation time and intraoperative bleeding volume in the group A were significantly more than those in the group B(P＜0.05),but there was no statistically significant difference in average hospitalization time between the two groups(P＞0.05).All cases were postoperatirely followed up for average (20.30-4-3.70) months.The levels of ESR and CRP,ODI score,Frankel score and Cobb angle after operation and at last follow up in the two group were significantly improved compared with before operation(P＜0.05).The bone graft healing time in the group A was significanlly shester than that in the group B.Conclusion Posterior approach fixation combined with anterior approach debridement operation has the advantages of less trauma,fixation stability and high bone healing rate for treating lumbar tuberculosis.

Objective To evaluate the diagnosis value of three-dimensional speckle tracking echocardiography ( 3D-STE) in severe multi-vessel coronary stenosis at resting state . Methods Ninety-one patients with chest tightness ,chest pain ,other related clinical symptoms ,and suspected coronary heart disease were enrolled and underwent coronary angiography ( CAG ) , two and three-dimensional echocardiography(2D-STE ,3D-STE) . According to CAG results ,the patients were divided into 3 groups:severe multi-vessel stenosis group ( n = 38 ) ,severe single-vessel stenosis group ( n = 22 ) ,and control group ( n =31) . All patients underwent two and three-dimensional echocardiography .3D-STE was applied to obtain left ventricular global longitudinal strain ( 3D GLS) ,global circumferential strain ( 3D GCS) ,global area strain (3D GAS) and global radial strain(3D GRS) . Receiver operating curve analysis was conduct to evaluate the diagnosis value of 3D-STE in severe multi-vessel coronary stenosis at resting state . Results 3D GLS ,3D GCS ,3D GAS and 3D GRS were decreased in the severe multi-vessel group and single-vessel stenosis group compared to those in the control group( P <0 .05) ;and greater decreases was observed in the severe multi-vessel stenosis group( P <0 .001) . Furthermore ,3D GLS and 3D GAS in the severe multi-vessel stenosis group were lower than those in severe single-vessel stenosis group( P <0 .05) . ROC curve showed that the cutoff value for 3D GLS was ≤ -11％ with a measurement sensitivity of 84 .2％ and cutoff value for 3D GAS was ≤ -20％ with a measurement specificity of 79 .3％ . A joint diagnostic test showed that the sensitivity was increased when 3D GLS was combined with at least one other parameters . Conclusions 3D-STE shows a great diagnostic potential when applied to detect severe multi-vessel coronary stenosis . Moreover ,GLS and GAS may act as more valuable indicators .

BACKGROUND:Vertebroplasty system has been proved to be effective for osteoporotic vertebral compressive fracture (OVCF); however, bone cement leakage-related complications occur frequently. Thereafter, high-viscosity bone cement system with high safety and efficacy is developed, but there is stil a lack of large-scale and high quality research. OBJECTIVE:To systematicaly review the efficacy and safety of high viscosityversus common bone cement systems for OVCF. METHODS: PubMed, Embase, Cochrane Library, CNKI, CqVip and WanFang databases were searched to colect the literatures about randomized controled trials (RCTs) or clinical controled trials (CCTs) of high viscosityversus common bone cement systems for OVCF published before January 2016. The quality assessment of included literatures and data extraction were performed by two researchers independently according to the Cochrane system. Data analysis was conducted using RevMan 5. 2 software. RESULTS AND CONCLUSION:Four RCTs and six CCTs were enroled. The results of Meta-analysis indicate that there were no significant differences in the volume of bone cement [MD=0.17, 95%CI(-0.04, 0.38)], visual analogue scale scores [RCT:MD=-0.30, 95%CI(-0.72, 0.11); CCT:MD=-0.05, 95%CI(-0.43, 0.32)] and incidence of second fractures [OR=1.77, 95%CI(0.24, 12.86)] between two bone cements. However, patients undergoing high viscosity bone cement system have significantly lower Oswestry disability index scores [MD=-2.99, 95%CI(-5.85,-0.13)], greater recovery of Cobb angle [MD=-3.19, 95%CI(-5.27,-1.10)] and lower incidence of cement leakage [OR=0.35, 95%CI(0.24, 0.51)]. High viscosity bone cement system in the treatment of OVCF shows good results at recovery of spinal structure and function and reducing leakage. Due to the limited quantity of included literature sand high heterogeneity, more large scale and high quality RCTs are stil needed for further verification.

Objective To explore the application value of platelet parameters detection in early diagnosis of breast cancer.Methods Platelet parameters of intravenous blood from 103 breast cancer patients and 117 fibroadenoma patients within 24 hours of admission between January 2015 and December 2015 were collected and retrospectively analyzed,including platelet count(PLT),plateletcrit(PCT),mean platelet volume(MPV),mean platelet component concentration(MPC),mean platelet component mass (MPM),platelet distribution width(PDW),and compared with control group of 100 healthy women.The diagnostic efficiency of platelet parameters were assessed using receiver operating characteristic(ROC) curve.Results PLT and PCT in breast cancer group and fibroadenoma group were significantly higher than those in control group(P＜0.01);MPC,PMM and PDW were significantly lower than those in control group(P＜0.01);MPV had no significant statistically difference(P＞0.05).The levels of MPC in breast cancer group were obviously lower than that in fibroadenoma group(P＜0.01).The diagnostic efficiency of MPC in breast cancer had the highest area under curve(AUC=0.835).While the cutoff value of MPC was 249.5 g/L,sensitivity and specificity were 82.5 ％ and 73.6 ％,respectively.Conclusion The detection of platelet parameters,especially MPC,have certain application value in early diagnosis of breast cancer.