Objective To evaluate the potential use of a probe melting analysis (PMA) assay in detecting the embB mutations which confer resistance against ethambutol in Mycobacterium tuberculosis. Methods The analysis sensitivity and specificity of PMA were investigated by detecting a serially diluted H37 Rv DNA and a reference panel from National Institute for the Control of Pharmaceutical and Biological Product. Six hundred and thirteen sputum samples were collected from the Xiamen Center for Disease Control and Prevention, Xiamen First Hospital and Center for Zhangzhou Disease Control and Prevention from September 2009 to April 2010. The PMA assay was then evaluated by detecting 613 clinical isolates and the results were compared with the sequencing results. Results The PMA assay could specifically detect Mycobacterium tuberculosis and had a limit of detection of 3 copies per reaction. The assay results with 613 clinical isolates showed that PMA gave a 100% concordance with sequencing in the 583 qualified samples, among which 34 were mutations at embB 306,23 at embB 378-380, 3 at embB 406 and 3 at embB 497. Conclusions PMA assay is a sensitive and specific method enabling efficient detection of common embB mutations causing ethambutol-resistance. The rapidness of this method together with its reliability would facilitate its use in routine testing.

The purpose of the present study was to investigate Norovirus infection status among patients with virus diarrhea in Xiamen city and provide evidence for exploring the prevalence characteristics and constituting appropriate control strategy.From April 2007 to July 2008,323 fecal samples of virus diarrhea cases collected from 3 surveillance hospitals in Xiamen were detected for antigen and RNA by ELISA and Real-Time RT-PCR respectively.The RdPd genes from some samples were furtherly amplified and sequenced for genogroup identification when the Real-Time RT-PCR detection results were positive.In the 323 fecal specimens,68 (21.05%) were positive for Norovirus antigen by ELISA and 107 (31.13%) were positive for Norovirus RNA by Real-Time RT-PCR.The overall positive prevalence rate of Norovirus in Xiamen was 38.08%.107 positive specimens were detected by Real-Time RT-PCR and results showed that 80 strains were Norovirus GGⅡ(74.77%),2 strains were GGⅠ(1.87%) and 25 strains (23.36%)were unidentified.It's indicated that Norovirus infection in Xiamen district was mainly caused by Norovirus GGⅡ,and Norovirus was also the main cause for virus diarrhea.

OBJECTIVETo develop a high-throughput rapid method for Vibrio (V.) cholerae molecular typing based on Melting Curve-based Multilocus Melt Typing (McMLMT).

METHODSSeven housekeeping genes of V.cholerae were screened out, and for each gene, the specific primers were designed for correspondent genes as well as 4 probes covering polymorphism loci of sequences. After optimizing all parameters, a method of melting-curve analysis following asymmetric PCR was established with dual-fluorescent-reporter in two reaction tubes for each gene. A set of 28 Tm-values was obtained for each strain and then translated into a set of code of allelic genes, standing for the strain's McMLMT type (MT). Meanwhile, sequences of the 7-locus polymorphism were typed according to the method of MLST. To evaluate the efficiency and reliability of McMLMT, the data were compared with that of sequence-typing and PFGE using BioNumerics software.

RESULTSMcMLMT method was established and refined for rapid typing of V. cholerae that a dozen of strains can be finished testing in a 3-hours PCR running using 96-well plates. 108 strains were analyzed and 28-Tm-values could be grouped and encoded according to 7 housekeeping gene to obtain the code set of allelic genes, and classified into 18 types (D = 0.723 3). Sequences of the 7 genes' polymorphism areas were directly clustered into the same 18 types with reference to MLST method. 46 of the strains, each represented a different PFGE type, could be classified into 13 types (D = 0.614 5) with McMLMT method and A- K groups at 85% similarity (D = 0.858 9) with PFGE method.

CONCLUSIONMcMLMT method is a rapid high-throughput molecular typing method for batches of strains with a resolution equal to MLST method and comparable to PFGE group.

Objective To investigate the effect of Shenqi Dihuang decoction combined with recombinant erythropoietin and L-carnitine on renal function in patients with end-stage renal disease and the regulation of Janus activated kinase/signal transducer and activator of transcriptions(JAK/STAT)pathway.Methods A total of 144 patients with end-stage renal disease who were treated in the Second Affiliated Hospital of Xingtai Medical College from October 2019 to October 2021 were selected as the study objects.Patients were divided into Shenqi Dihuang decoction treatment group and conventional treatment group according to random number table method,72 cases in each group.Maintenance hemodialysis was performed in both groups.The conventional treatment group received recombinant erythropoietin and L-carnitine,while the Shenqi Dihuang decoction treatment group had Shenqi Dihuang decoction(prescription composition:30 g each of raw Astragalus,Parasitic loranthus,Eclipta,Polyporus,Poria bark,Raw coix seed,Salvia miltiorrhiza,and Pyrrosia,15 g each of Codonopsis pilosula,Dogwood,Herba lycopi,and Common yam rhizome,10 g each of raw Rehmannia glutinosa,Litchi core,Silkworm sand and Curcuma zedoary,6 g Cassia seed),once a day for 3 months.The clinical efficacy,renal function,microinflammatory status and serum JAK/STAT pathway related protein levels were observed after treatment in two groups of patients with different treatment modalities,and the occurrence of adverse reactions was recorded.Results The total effective rate of Shenqi Dihuang decoction was higher than that of conventional treatment group[90.28%(64/72)vs.77.78%(55/72),P<0.05].After treatment,the residual renal function(RRF),24-hour urine protein,blood urea nitrogen(BUN),blood creatinine(SCr)and inflammatory factors[hypersensitive C-reactive protein(hs-CRP),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)]were lower than before treatment,the post-treatment RRF of the Shenqi Dihuang decoction treatment group was higher than that of the conventional treatment group(mL/min:4.82±1.18 vs.3.96±1.05),while 24 hours urine protein(mg:62.26±12.16 vs.97.71±16.28),BUN(mmol/L:16.25±3.64 vs.20.65±4.13),SCr(μmol/L:242.25±25.62 vs.280.62±26.63),hs-CRP(mg/L:5.86±1.15 vs.7.78±1.32),IL-6(ng/L:3.26±0.64 vs.4.62±1.13)and TNF-α(μg/L:29.23±5.64 vs.32.66±6.13)were significantly lower than those in the conventional treatment group(all P<0.05).After treatment,JAK and STAT in Shenqi Dihuang decoction group were increased compared with before treatment,phosphorylated JAK(p-JAK)and phosphorylated STAT(p-STAT)were decreased compared with before treatment(both P<0.05),while the serum JAK/STAT pathway related protein levels in conventional treatment group were not significantly changed(all P>0.05).Therefore,JAK and STAT in the Shenqi Dihuang decoction treatment group after treatment were significantly higher than those in the conventional treatment group[JAK(μg/L):1.46±0.28 vs.1.26±0.26,STAT(μg/L):1.37±0.25 vs.0.99±0.24,both P<0.05],p-JAK and p-STAT were significantly lower than those in the conventional treatment group[p-JAK(μg/L):0.45±0.08 vs.0.65±0.13,p-STAT(μg/L):0.66±0.13 vs.0.82±0.28,both P<0.05].There was no statistically significant difference in the incidence of adverse reactions between the two groups[13.88%(10/72)vs.9.72%(7/72),P>0.05].Conclusion Taking Shenqi Dihuang decoction based on recombinant erythropoietin and L-carnitine treatment can effectively inhibit the JAK/STAT signaling pathway,and improve renal function and micro-inflammatory status in patients with end-stage renal disease,thereby improving the therapeutic effect.

Objective To investigate the optimum reperfusion duration of liver ischemia-reperfusion injury (IRI) model by occluding the proper hepatic artery in rats. Methods Fourty-eight Sprague Dawley (SD) rats with mean weight of (200±25) g were randomly assigned to 8 groups by random number table method:IRI-0, 1, 3, 6, 12, 24, 48 h and sham operation (SO) group with 6 rats in each group. The proper hepatic arteries in rats of IRI groups were selectively occluded for 1 h and then blood lfow recovered. Samples of blood and liver tissues were collected at the time points of 0, 1, 3, 6, 12, 24, 48 h of reperfusion. In SO group, samples were collected after the proper hepatic artery was isolated and the ifrst portal was exposed for 1 h. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), pathological changes, DNA fragmentation rates, and levels of Ki-67 expression of liver tissues were observed in each group. Measurement data of multiple groups were compared using one-way analysis of variance and LSD-t test. Results The levels of serum ALT in IRI-0, 1, 3, 6, 12, 24, 48 h and SO group were (53±25), (85±20), (96±18), (411±96), (87±19), (81±15), (46±6), (60±14) U/L respectively. The levels of serum AST were (238±63), (364±111), (375±68), (1 291±511), (800±87), (854±218), (484±219), (248±94) U/L accordingly. The levels of serum ALT, AST in IRI 6 h group were the highest (F=36.015, 18.241;P<0.05). The damage of liver tissues in IRI 6 h group was the most serious. The DNA fragmentation rates of liver tissues were (7.5±1.5)%, (9.2±2.2)%, (9.3±2.3)%, (12.6±2.4)%, (6.3±1.0)%, (5.4±0.9)%, (4.5±0.8)%, (4.5±1.1)%accordingly, which was the highest in IRI 6 h group (F=15.992, P<0.05). The levels of Ki-67 expression of liver tissues were (3.5±1.4), (5.6±1.8), (8.7±2.3), (13.7±2.4), (15.2±1.2), (20.5±2.2), (31.8±2.5), (2.4±1.2)/high power ifeld accordingly, which was the highest in IRI 48 h group (F=261.707, P<0.05). Conclusions The liver IRI model can be successfully established by occluding the proper hepatic artery, and the optimum reperfusion duration of IRI is 6 h.

Objective:To investigate the clinical utility of intracavitary electrocardiogram (IC-ECG) localization technique in locating the tip of umbilical venous catheterization (UVC).Methods:From April 2020 to March 2021, premature infants admitted to the Department of Neonatology of the Third Affiliated Hospital of Guangzhou Medical University and received UVC were collected for a prospective study. According to whether IC-ECG was used to determine the position of umbilical vein catheter tip during catheterization, they were divided into IC-ECG group and control group. The differences of catheterization success rate, total incidence of catheter-related complications were compared between the two groups. The relationship between characteristic P waves and the position of the catheter tip in the IC-ECG group was also analyzed.Results:A total of 104 premature infants were enrolled, including 52 cases in IC-ECG group and 52 cases in control group. The catheter placement accuracy rate of the IC-ECG group was higher than that of the control group (53.8% vs. 28.8%, P<0.05). There was no statistically significant difference of the total incidence of catheter-related complications between the two groups ( P>0.05). Among the 40 cases with characteristic P wave changes in the IC-ECG group, 21 cases had normal P wave, of which 13 cases (61.9%) were with the correct position. And 19 cases had low-amplitude bimodal P waves, of which 15 cases (78.9%) were with the correct position. There was no statistically significant difference of the exact position rate of the catheter tips between the two types of P waves ( P>0.05). Conclusions:The use of IC-ECG localization technique in the process of catheterization of umbilical vein can increase the exact position rate of catheterization. Furthermore, when IC-ECG monitoring shows characteristic P wave changes to normal P wave or low-amplitude bimodal P wave, the tip of the UVC catheter may be located accurately.

Objective:To analysis effectiveness of the "14 plus 7 day quarantine" and "nucleic acid plus total antibody testing" strategy (combined screening strategy) for screenin the imported patients with COVID-19 in Xiamen.Methods:The study populations were overseas travelers arriving in Xiamen from March 17 to December 31, 2020, and overseas travelers who had quarantine outside Xiamen for less than 21 days from July 18 to December 31, 2020. Data were collected and analyzed on the timing of detection, pathways, and test results of the imported patients with COVID-19 after implementing combined screening strategy.Results:A total of 304 imported patients with COVID-19 were found from 174 628 overseas travelers and 943 overseas travelers from other cities. A total of 163 cases (53.6%) were diagnosed by multitime, multisite intensive nucleic acid testing after positive finding in total antibody testing. Among them, 27 (8.9%) were first positive for nucleic acid in 14 plus 7 day quarantine and 136 were first positive for nucleic acid in 14-day quarantine. Only 8 of these individuals were tested positive for nucleic acid after positive total antibody testing. The other 128 individuals were tested positive for nucleic acid after being negative for average 2.3 times (maximum of 6 times). Aditional 155 cases might be detected by using the combined "14 plus 7 day quarantine" and " nucleic acid plus total antibody testing" strategy compared with "14-day quarantine and nucleic acid testing" strategy, accounting for 51.0% of the total inbound infections. So the combined screening strategy doubled the detection rate for imported patients with COVID-19. No second-generation case caused by overseas travelers had been reported in Xiamen as of February 26, 2021.Conclusions:Xiamen's combined screening strategy can effectively screen the imported patients with COVID-19 who were first positive for nucleic acid after 14 day quarantine. Compared with "14 day quarantine and nucleic acid testing", the combined screening strategy improved detection rate and further reduced the risk of the secondary transmission caused by the imported patients with COVID-19.

Background::Cardiovascular disease (CVD) has emerged as the leading cause of death from prostate cancer (PCa) in recent decades, bringing a great disease burden worldwide. Men with preexisting CVD have an increased risk for major adverse cardiovascular events when treated with androgen deprivation therapy (ADT). The present study aimed to explore the prevalence and risk evaluation of CVD among people with newly diagnosed PCa in China.Methods::Clinical data of newly diagnosed PCa patients were retrospectively collected from 34 centers in China from 2010 to 2022 through convenience sampling. CVD was defined as myocardial infarction, arrhythmia, heart failure, stroke, ischemic heart disease, and others. CVD risk was estimated by calculating Framingham risk scores (FRS). Patients were accordingly divided into low-, medium-, and high-risk groups. χ2 or Fisher’s exact test was used for comparison of categorical variables. Results::A total of 4253 patients were enrolled in the present study. A total of 27.0% (1147/4253) of patients had comorbid PCa and CVD, and 7.2% (307/4253) had two or more CVDs. The enrolled population was distributed in six regions of China, and approximately 71.0% (3019/4253) of patients lived in urban areas. With imaging and pathological evaluation, most PCa patients were diagnosed at an advanced stage, with 20.5% (871/4253) locally progressing and 20.5% (871/4253) showing metastasis. Most of them initiated prostatectomy (46.6%, 1983/4253) or regimens involving ADT therapy (45.7%, 1944/4253) for prostate cancer. In the present PCa cohort, 43.1% (1832/4253) of patients had hypertension, and half of them had poorly controlled blood pressure. With FRS stratification, as expected, a higher risk of CVD was related to aging and metabolic disturbance. However, we also found that patients with treatment involving ADT presented an originally higher risk of CVD than those without ADT. This was in accordance with clinical practice, i.e., aged patients or patients at advanced oncological stages were inclined to accept systematic integrative therapy instead of surgery. Among patients who underwent medical castration, only 4.0% (45/1118) received gonadotropin releasing hormone antagonists, in stark contrast to the grim situation of CVD prevalence and risk.Conclusions::PCa patients in China are diagnosed at an advanced stage. A heavy CVD burden was present at the initiation of treatment. Patients who accepted ADT-related therapy showed an original higher risk of CVD, but the awareness of cardiovascular protection was far from sufficient.