Objective: To investigate the environmental contamination of norovirus in nurseries and primary/secondary schools, so as to provide a scientific basis for effective prevention and control measures. Methods: A total of 483 external environmental samples were collected from 34 cluster outbreaks of norovirus gastroenteritis in kindergartens and primary/secondary schools in Linhai City from 2021 to 2024. Pathogen detection was conducted using a rapid nucleic acid extraction kit and realtime fluorescence RT-PCR, and the results were analyzed using the χ2 test or Fishers exact test. Results: Among the collected external environmental samples, the total positive rate of surface contamination was 13.66%. The positive rates in kindergartens and primary/secondary schools were 12.20% and 15.82%, respectively. In kindergartens, the five surfaces with the highest detection rates were desks/chairs (23.33%), toilet stool troughs (20.69%), urinal troughs (12.00%), washbasins/sinks (11.11%), and toilet mops (9.38%). In primary/secondary schools, the top five were toilet stool troughs (38.30%), urinal troughs (23.53%), toilet door handles (13.04%), toilet mops (12.50%), and drinking cups (11.11%). The difference in positive detection rates among different external environments in primary/secondary schools was statistically significant (Fishers exact probability test, P<0.01). The positive detection rate in sanitary toilets was higher than that in classroom environments (χ2=17.38), while the positive detection rate in classroom environments of kindergartens was higher than that in primary/secondary schools (χ2=5.42)(P<0.05). Conclusions Norovirus exhibits a high contamination rate in nurseries and schools, particularly in restroom areas. Strengthening sanitation and disinfection in highrisk environments, and improving hygiene awareness among children and staff, are essential for the effective prevent and control of norovirus.

Objective To investigate the expression of YWHAQ protein in gastric adenocarcinoma tissues and its correlation with clinical pathological features and prognosis. Methods A total of 127 patients with gastric cancer who underwent radical surgery were enrolled. Clinical data and postoperative cancer tissue samples were collected from the patients. Immunohistochemistry was used to detect the protein expression of YWHAQ in gastric adenocarcinoma tissues. The relationship between YWHAQ expression and clinical pathological features and prognosis was analyzed. Bioinformatics prediction was performed to identify potential pathways regulated by YWHAQ in gastric adenocarcinoma. A protein-protein interaction network for YWHAQ was constructed using the STRING database. Results YWHAQ gene expression was significantly higher in gastric adenocarcinoma tissues than in normal tissues (P<0.05). The expression level of the YWHAQ protein was significantly correlated with age, tumor invasion depth, lymph node metastasis, and tumor stage (P<0.05). Kaplan-Meier survival analysis showed that patients with high YWHAQ expression had significantly poorer long-term survival than those with low expression (P<0.0001). Cox regression analysis revealed that high YWHAQ expression and distant metastasis were independent risk factors for gastric cancer (HR>1, P<0.05). Enrichment analysis indicated that YWHAQ positively regulated processes such as cell cycle, angiogenesis, and DNA damage repair. Conclusion YWHAQ is highly expressed in gastric adenocarcinoma tissues and may be associated with the occurrence, invasion, and metastasis of gastric adenocarcinoma. High YWHAQ expression and distant metastasis are independent risk factors for the prognosis of gastric adenocarcinoma patients.

BACKGROUND:Apoptosis and autophagy imbalance in the hippocampal region of perimenopausal depressed rats are closely related to cognitive decline.Whether aerobic exercise can reduce apoptosis by promoting hippocampal autophagy and thus improve the learning and memory abilities of perimenopausal depressed rats is not clear.OBJECTIVE:To investigate the possible mechanism by which 4-week moderate-intensity treadmill exercise improves learning memory ability in ovariectomized stressed rats.METHODS:Forty Sprague-Dawely rats were randomly divided into four groups,namely,sham operation group(n=10),ovariectomized group(n=10),ovariectomized stress group(n=10)and ovariectomized stress exercise group(n=10).Except for the sham operation group,the ovaries were removed in the other three groups to establish a perimenopausal rat model,and then a depressed rat model was established by chronic unpredictable stress in the latter two groups.The rats in the ovariectomized stress exercise group underwent a 4-week moderate-intensity treadmill exercise.Tail suspension test and sucrose preference test were performed to text depression-like behaviors in rats after exercise and stress.The eight-arm maze experiment was used to test the learning and memory behaviors of rats after exercise and stress.Western blot was used to detect the protein levels of AMP-activated protein kinase/UNC-51 like autophagy activating kinase 1/mammalian target of rapamycin(AMPK/mTOR/ULK1),hippocampus apoptotic factor Caspase-3 and the protein expression of autophagy markers LC-3II/Beclin-1 in the hippocampus.RESULTS AND CONCLUSION:(1)Compared with the sham operation group,rats in the ovariectomized and ovariectomized stress groups had prolonged resting time in the tail suspension test and decreased sugar-water intake and sugar-water preference in the sucrose preference test.(2)Ovary removal reduced the learning memory capacity of rats,as evidenced behaviorally by a significant increase in the number of working memory errors,the number of reference memory errors,and the completion time,and an even more pronounced increase in the above measures in the ovariectomized stress group.(3)Compared with the ovariectomized group,there was a significant reduction in the number of working memory errors,the number of reference memory errors,and the completion time in the ovariectomized stress group.(4)Compared with the sham operation group,in the ovariectomized and ovariectomized stress groups,the expression of hippocampal apoptotic factor Caspase 3 protein was significantly elevated,the expression of autophagy-related factors proteins Beclin-1 and LC3II,as well as the protein expression of AMPK and ULK1,was decreased,whereas the expression of mTOR protein was elevated.Changes in the above indicators were more significant in the ovariectomized stress group.(5)Compared with the ovariectomized stress group,in the ovariectomized stress exercise group,the protein expression of Caspase 3 was significantly decreased,the protein expression of Beclin-1 and LC3II was significantly increased,the protein expression of AMPK and ULK1 was significantly increased,and the protein expression of mTOR was significantly reduced.To conclude,4-week moderate-intensity treadmill exercise may promote cellular autophagy and reduce apoptosis through the AMPK/mTOR/ULK1 autophagy signaling pathway,thereby enhancing the learning and memory capacity of rats with ovariectomized depression

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective To explore the methods and contents of clinical microbiology testing in the"Sansheng"teaching practice in the information age,and guide the cultivation of high-level laboratory technology professionals with comprehensive quality.Methods Through thinking and improving the practice teaching mode of the laboratory department of Qingyuan Hospital Affiliated to Guangzhou Medical University,using the existing curriculum design,combined with the"Internet+education"back-ground,let"Sansheng"have a deeper understanding and application in the study of microbial testing.Results Ideological and po-litical education runs through the whole process of medical practice,which helps to cultivate students'sense of professional identi-ty,belonging and social mission;In the teaching process,the dual planning teaching mode of case-based learning and problem-based learning was introduced.At the same time,the"Internet+"technology was used to carry out diversified teaching.Com-bined with clinical practice,students'ability to analyze and solve problems was significantly improved,and the goal of quality edu-cation was achieved.Conclusion The teaching application of clinical microbiological examination based on the"Internet+"mode can effectively improve the teaching quality,improve the comprehensive ability of students and improve the learning effect.

BACKGROUND:Apoptosis and autophagy imbalance in the hippocampal region of perimenopausal depressed rats are closely related to cognitive decline.Whether aerobic exercise can reduce apoptosis by promoting hippocampal autophagy and thus improve the learning and memory abilities of perimenopausal depressed rats is not clear.OBJECTIVE:To investigate the possible mechanism by which 4-week moderate-intensity treadmill exercise improves learning memory ability in ovariectomized stressed rats.METHODS:Forty Sprague-Dawely rats were randomly divided into four groups,namely,sham operation group(n=10),ovariectomized group(n=10),ovariectomized stress group(n=10)and ovariectomized stress exercise group(n=10).Except for the sham operation group,the ovaries were removed in the other three groups to establish a perimenopausal rat model,and then a depressed rat model was established by chronic unpredictable stress in the latter two groups.The rats in the ovariectomized stress exercise group underwent a 4-week moderate-intensity treadmill exercise.Tail suspension test and sucrose preference test were performed to text depression-like behaviors in rats after exercise and stress.The eight-arm maze experiment was used to test the learning and memory behaviors of rats after exercise and stress.Western blot was used to detect the protein levels of AMP-activated protein kinase/UNC-51 like autophagy activating kinase 1/mammalian target of rapamycin(AMPK/mTOR/ULK1),hippocampus apoptotic factor Caspase-3 and the protein expression of autophagy markers LC-3II/Beclin-1 in the hippocampus.RESULTS AND CONCLUSION:(1)Compared with the sham operation group,rats in the ovariectomized and ovariectomized stress groups had prolonged resting time in the tail suspension test and decreased sugar-water intake and sugar-water preference in the sucrose preference test.(2)Ovary removal reduced the learning memory capacity of rats,as evidenced behaviorally by a significant increase in the number of working memory errors,the number of reference memory errors,and the completion time,and an even more pronounced increase in the above measures in the ovariectomized stress group.(3)Compared with the ovariectomized group,there was a significant reduction in the number of working memory errors,the number of reference memory errors,and the completion time in the ovariectomized stress group.(4)Compared with the sham operation group,in the ovariectomized and ovariectomized stress groups,the expression of hippocampal apoptotic factor Caspase 3 protein was significantly elevated,the expression of autophagy-related factors proteins Beclin-1 and LC3II,as well as the protein expression of AMPK and ULK1,was decreased,whereas the expression of mTOR protein was elevated.Changes in the above indicators were more significant in the ovariectomized stress group.(5)Compared with the ovariectomized stress group,in the ovariectomized stress exercise group,the protein expression of Caspase 3 was significantly decreased,the protein expression of Beclin-1 and LC3II was significantly increased,the protein expression of AMPK and ULK1 was significantly increased,and the protein expression of mTOR was significantly reduced.To conclude,4-week moderate-intensity treadmill exercise may promote cellular autophagy and reduce apoptosis through the AMPK/mTOR/ULK1 autophagy signaling pathway,thereby enhancing the learning and memory capacity of rats with ovariectomized depression

Objective To explore the methods and contents of clinical microbiology testing in the"Sansheng"teaching practice in the information age,and guide the cultivation of high-level laboratory technology professionals with comprehensive quality.Methods Through thinking and improving the practice teaching mode of the laboratory department of Qingyuan Hospital Affiliated to Guangzhou Medical University,using the existing curriculum design,combined with the"Internet+education"back-ground,let"Sansheng"have a deeper understanding and application in the study of microbial testing.Results Ideological and po-litical education runs through the whole process of medical practice,which helps to cultivate students'sense of professional identi-ty,belonging and social mission;In the teaching process,the dual planning teaching mode of case-based learning and problem-based learning was introduced.At the same time,the"Internet+"technology was used to carry out diversified teaching.Com-bined with clinical practice,students'ability to analyze and solve problems was significantly improved,and the goal of quality edu-cation was achieved.Conclusion The teaching application of clinical microbiological examination based on the"Internet+"mode can effectively improve the teaching quality,improve the comprehensive ability of students and improve the learning effect.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective:To investigate the effects of thinned anterolateral thigh perforator flaps combined with finger splitting and webplasty in sequential treatment of degloving destructive wound of total hand.Methods:This study was a retrospective observational study. From January 2012 to January 2023, a total of 15 cases who met the inclusion criteria with degloving destructive wound of total hand were admitted to Ningbo No.6 Hospital, including 10 males and 5 females, aged 17-75 years. The wounds were all combined with exposed bones or tendon. Emergency debridement and vacuum sealing drainage were performed in all cases before flap transplantation in stage Ⅰ. After thorough debridement, the wound area was 11.0 cm×3.0 cm-23.0 cm×13.5 cm. One or both anterolateral thigh perforator flaps with size of 12.5 cm×5.0 cm-25.0 cm×15.5 cm were designed, cut, and thinned to repair the skin and soft tissue defects of the hand. The donor site was sutured directly or repaired with medium-thickness skin graft from the opposite thigh. As needed, the flap was reconstructed by finger splitting and webplasty once or more times every 3 months after stage Ⅰoperation. The survival and complications of flap and wound healing at the donor site were observed after stage Ⅰoperation. The appearance of flap, two-point discrimination distance, and hand function were observed during the follow-up. At the final follow-up, the function of the affected hand was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association.Results:After the operation of stage Ⅰ, all the flaps of 15 cases of patients survived completely, including 1 case that had arterial crisis of flap but survived completely after exploration and re-anastomosis of blood vessels; all the wounds at the donor site healed. During the follow-up period of 6 to 18 months after stage Ⅰ, the flap was slightly swollen, with a little pigmentation, and the two-point discrimination distance in the finger flap was 8-11 mm. The fingers could complete the basic life actions such as flexion, extension, pinch, and grip. At the final follow-up, 3 cases were excellent, 9 cases were good, and 3 cases were acceptable in function evaluation of the affected hand.Conclusions:For degloving destructive wound of total hand, free transplantation of one or both thinned anterolateral thigh perforator flaps is used for repair in stage Ⅰ, and finger splitting and webplasty are used to reconstruct the flaps in the later stage, which can basically restore the pinch and grip function of the affected hand that is required for daily life, and is worthy of clinical promotion.

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.