Objective To establish a fluorescent recombinase-aided amplification (RAA) assay for detection of Strongyloides stercoralis nucleic acid and to preliminarily evaluate its performance. Methods Six sets of specific primers targeting S. stercoralis 18S ribosomal RNA (18S rRNA) gene and one fluorescent probe were designed and synthesized. The optimal primer-probe set was determined through systematic screening and optimization to establish the fluorescent RAA assay. The assay was evaluated using S. stercoralis genomic DNA at concentrations of 100, 10, and 1 pg/μL, and 100, 10, and 1 fg/μL, as well as recombinant pUC57 plasmids containing the target gene fragments at 1 × 10⁵, 1 × 10⁴, 1 × 10³, 1 × 10², 1 × 10¹, 1 × 10⁰ copies/reaction, to determine the analytical sensitivity. Genomic DNA from Ascaris lumbricoides, Ancylostoma duodenale, Enterobius vermicularis, Angiostrongylus cantonensis, Trichinella spiralis, Clonorchis sinensis, Schistosoma japonicum, and Taenia saginata was used to assess assay specificity. A total of 25 stool samples from patients suspected of S. stercoralis infection were tested by the modified Baermann funnel technique, PCR, and the established fluorescent RAA assay. The sensitivity, specificity, concordance rate and their 95% confidence intervals (CI) of these three techniques were estimated, and agreement between methods was evaluated using the Kappa coefficient. Results Exo-4 was identified as the optimal primer set screened from the six primer sets, and the best amplification performance was achieved when the final concentrations of the forward and reverse primers were 0.44 μmol/L and a probe concentration was 0.20 μmol/L. The limit of detection of the fluorescent RAA assay was 100 fg/μL for genomic DNA of S. stercoralis and 1 × 10⁰ copies/reaction for recombinant plasmids. Specific fluorescence signals were detected within 5 min, with no cross-reactivity observed with A. lumbricoides, A. duodenale, E. vermicularis, A. cantonensis, T. spiralis, C. sinensis, S. japonicum, or T. saginata. Among the 25 clinical stool samples from patients suspected of S. stercoralis infections, the modified Baermann funnel technique and fluorescent RAA assay detected 19 positives and 6 negatives, whereas PCR detected 18 positives and 7 negatives. The fluorescent RAA assay showed a sensitivity of 100.00% [95% CI: (82.35%, 100.00%)], specificity of 100.00% [95% CI: (54.07%, 100.00%)], concordance rate of 100.00% [95% CI: (86.28%, 100.00%)], and a Kappa coefficient of 1.00 [95% CI: (1.00, 1.00)] (P < 0.001) relative to the modified Baermann funnel technique, and a sensitivity of 100.00% [95% CI: (81.47%, 100.00%)], specificity of 85.71% [95% CI: (42.13%, 99.64%)], concordance rate of 96.00% [95% CI: (79.65%, 99.90%)], and a Kappa coefficient of 0.90 [95% CI: (0.70, 1.00)] (P < 0.001). Positive amplification products emitted green fluorescence under a portable blue-light device, enabling visual interpretation of results. Conclusions The fluorescent RAA assay established in this study is rapid, highly sensitive, and highly specific. It enables detection of S. stercoralis nucleic acid under isothermal conditions and allows visual interpretation of results, providing a novel tool for rapid clinical diagnosis and field screening of S. stercoralis infections.

Objective To observe the effect of the three active ingredients of a Chinese traditional medicine compound named Kang Fu Ling( KFL) against PC12 cells oxidative damage induced by microwave radiation.Methods PC12 cells were differentiated into neuros induced by nerve growth factor ( NGF ) .PC12 cells were incubated for 48 hours after astragalosides,total paeony glycoside and tanshinones were added at different concentrations (1, 3, or 9 μg/ml) .The cells in the control group were cultivated with the only medium of the same volume.Then, cells were irradiated with 30 mW/cm2 microwave for 6 minutes.The morphology of PC12 cells was observed under an inverted microscope soon before and after irradiation and the cell viability was measured by methylthiazolyl tetrazolium ( MTT) colorimetry.Reactive oxygen species ( ROS ) was determined using active oxygen probe 2′, 7′-dichlorodihyarofluolescen diacetde ( DCFH-DA ) while malonyldialdehyde(MDA) was measured in the homogenate of PC12 cells through thiobarbituric acid ( TBA) reactive substance assay.Results The cell morphology of each group showed no obvious difference.6 h after irradiation, the viability of irradiation control group measured by MTT declined apparently(P<0.01)compared with the normal control group.The 3 μg/ml astragalosides treatment group increased the viability of PC12 cells after microwave exposure ( P <0.01).The contents of ROS and MDA were increased after irradiation(P<0.01).However, in the three active ingredients of Kang Fu Ling treatment groups, both ROS and MDA were much lower than in irradiation control group.Conclusion Astragalosides, total paeony glycoside and tanshinones, which are the three active ingredients of Kang Fu Ling, all have protective effect against PC12 cell injury caused by microwave radiation,possibly by scavenging free radicals and reducing oxidative stress injury.

ObjectiveTo investigate the control rate and influence factors of glycolated hemoglobin A1C(HbA1c) of type 2 diabetes in Baoding city community.MethodsA cluster-randomized study was conducted and three communities were selected randomly.The study involved all of people aged 45 years old and above in the three communities.The type 2 diabetic patients diagnosed before,were recorded through the cross section study.Then the questionnaire was finished.All diabetic plasma HbA1c was determined by HPLC method.ResultsEighty-seven patients with HbA1c≤7％ were only 18.6％ in all diabetic patients.As the plasma level of HbA1c increased,occurrences of macroangiopathy and microangiopathy were both increased,by the trend test.Logistic regression analysis showed that therapeutic measures and knowledge about diabetes were main influence factors of achieving the hemoglobin Alc target of ＜7％ in type 2 diabetes.ConclusionThere were low HbA1c control in diabetic patients of Baoding community,and knowing diabetes well and receiving insulin treatment might decrease HbA1c level apparently.

Objective To explore the characteristic findings of 1H-MR spectroscopy(1H-MRS) in the prefrontal cortex and amygdala of patients with heroine dependence(HD), and the relationship to total cumulative dose of inhaled heroine.Methods Fourteen male HD patients and 12 healthy controls (HC) underwent 1H-MRS at the prefrontal cortex and amygdala regions.The total cumulative in haled heroin dose was (852±341) g in HD.Ratios of N-acetylaspartate/creatine(NAA/Cr) and choline/creatine (Cho/Cr) were respectively measured in the prefrontal cortex and bilateral amygdale regions.The student's t test and the linear correlation were employed for statistical analysis.Results Compared to HC group, HD patients had a significant lower ratio of NAA/Cr in the prefrontal cortex (1.44±0.46 vs 1.50±0.75, t=1.77 ,P <0.05), left amygdala region (1.32±0.08 vs 1.42±0.08, t=3.41, P < 0.05), and right amygdala region (1.34±0.09 vs 1.44±0.10, t=2.63, P <0.05), the HD patients had a significant increased ratio of Cho/Cr in the prefrontal cortex (0.92±0.06 vs 0.86±0.08, t=2.31, P < 0.05), left amygdala region (1.20+0.12 vs 1.07±0.04, t=3.60,P<0.05) and right amygdala region(1.26±0.15 vs 1.12±0.11,t=2.60,P <0.05).There was a negative linear correlation between the total cumulative inhaled heroine dose and the ratio of NKA/Cr in the prefrontal cortex (r=-0.9159, P < 0.01), left amygdala region (r=-0.8756, P < 0.01), and right amygdala region (r=-0.9399, P < 0.01) respectively.Conclusions The study indicates that neuronal damage and glial proliferation may occur in the prefrontal cortex and amygdala region, which suggests the abnormalities of executive function and emotion in patients with HD.A relationship exists between the heroin-induced metabolic abnormality and the total cumulative dose of inhaled heroine.

OBJECTIVETo summarize retrospectively the 5-year survival rates and long-term complication of stage Ib-IIIa cervical carcinoma treated by combination of subradical external radiation and brachytherapy plus radical operation.

METHODS106 patients with cervical carcinoma were all treated by radical hysterectomy and pelvic lymphadenectomy, of whom 78 had had preoperative radiotherapy (external radiation and brachytherapy), 16 combination of brachytherapy and radical operation, 12 adjuvant postoperative radiotherapy (external radiation and brachytherapy). (60)Co was used for external radiation, in which the point B dose was 25 - 30 Gy in preoperative radiation and 40 - 50 Gy in postoperative radiation. (192)Ir high-dose-rate afterloading unit was used in brachytherapy, with a dose of 6 - 18 Gy at point A.

RESULTSThe follow up rate was 95.3%. The overall 5-year survival rates were 78.2% (61/78) in the preoperative radiotherapy group, 68.8% (11/16) in brachytherapy plus radical operation, 33.3% (4/12) in the postoperative radiotherapy group, showing a higher 5-year survival rate in the preoperative radiotherapy group than the postoperative radiotherapy group (P < 0.05). In stage II patients, the preoperative radiotherapy group -77.6% (45/58) also gave a higher survival than the postoperative radiotherapy group -25.0% (1/4) (P < 0.05). But all the other groups gave differences of insignificance. The chief long-term complications were radio-proctitis and cystitis, with incidences of 34.6% (27/78), 31.3% (5/16), 33.3% (4/12) in the preoperative radiotherapy group, brachytherapy plus radical operation group and the postoperative radiotherapy group (P > 0.05).

CONCLUSIONThe overall 5-year survival rate of combined subradical external radiation and brachytherapy plus radical operation was obviously higher than that of postoperative radiotherapy for stage Ib-IIIa and II patients, with statistically significant differences. However, the incidence of long-term complications give no statistical significance in the preoperative radiotherapy group or brachytherapy plus the operation group as compared with the postoperative radiotherapy group.

Adult ; Aged ; Aged, 80 and over ; Brachytherapy ; Combined Modality Therapy ; Female ; Humans ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Rate ; Uterine Cervical Neoplasms ; mortality ; radiotherapy ; surgery