ObjectiveTo establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus (MHV), mouse parvovirus (MPV), and reovirus type 3 (Reo-3) in laboratory mice. MethodsRecombinant plasmid standards were constructed using conserved genomic regions of the three target viruses. Specific primers and probes were designed, and a triplex TaqMan quantitative PCR (qPCR) system was optimized through preliminary experiments. Sensitivity was evaluated using 10-fold serial dilutions (10¹-10⁷ copies/μL). Intra- and inter-assay repeatability were assessed via reproducibility experiments. ResultsA strong linear correlation (R ²>0.99) was observed between copy number and Ct value at template concentrations of 10²-10⁷ copies/μL. Intra-assay and inter-assay coefficients of variation (CV) were both below 5%. No cross-reactivity was observed with mouse cytomegalovirus (MCMV), vesicular stomatitis virus (VSV), Sendai virus (SeV), or pneumonia virus of mice (PVM). ConclusionThe triplex TaqMan quantitative PCR method established in this study has high sensitivity, good repeatability, and strong specificity. It enables the rapid, specific, and sensitive detection of MHV, MPV, and Reo-3, allows the simultaneous detection of multiple pathogens in a single tube, and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations, thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective:To evaluate the reliability and validity of the Mandarin version of the patient-rated tennis elbow evaluation(PRTEE)for patients with tennis elbow.Method:With the permission of the author of the original questionnaire,the PRTEE was translated into Chi-nese following current international procedures.A total of 100 subjects were included in the study,and were assessed using the visual analogue scale(VAS),the Disabilities of the Arm,Shoulder,and Hand(DASH)ques-tionnaire,and PRTEE.A second assessment using the PRTEE was completed with 75 participants after one week.The internal consistency of the scale was evaluated using Cronbach's alpha coefficient,and the test-re-test reliability was evaluated using the intraclass correlation coefficient(ICC).The construct validity of the Mandarin version of PRTEE scale was examined by analyzing correlation with the Chinese version of VAS and DASH scores using Pearson's correlation coefficient.The ceiling or floor effect was determined by explor-atory analysis.Result:The Cronbach's alpha coefficient of PRTEE total score,pain and function subscales were 0.947,0.872,and 0.927,respectively.The ICC values for test-retest reliability of PRTEE total score,pain,and func-tion subscales were 0.988、0.975,and 0.980,respectively.The correlation coefficient between PRTEE total score and its two subscales and VAS ranged from 0.838 to 0.920(P＜0.01),indicating a strong correlation.The correlation coefficient between the PRTEE total score and two subscales,the DASH total score and two di-mensions ranged from 0.704 to 0.893(P＜0.01),indicating a strong correlation.There were no subjects who re-ceived the lowest or highest scores,indicating that the scale had no ceiling or floor effects.Conclusion:The Mandarin version of PRTEE demonstrates good reliability and validity,with no evidence of ceiling or floor effect.It is suitable for evaluating tennis elbow in Mandarin-speaking patients.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Objective:To evaluate the reliability and validity of the Mandarin version of the patient-rated tennis elbow evaluation(PRTEE)for patients with tennis elbow.Method:With the permission of the author of the original questionnaire,the PRTEE was translated into Chi-nese following current international procedures.A total of 100 subjects were included in the study,and were assessed using the visual analogue scale(VAS),the Disabilities of the Arm,Shoulder,and Hand(DASH)ques-tionnaire,and PRTEE.A second assessment using the PRTEE was completed with 75 participants after one week.The internal consistency of the scale was evaluated using Cronbach's alpha coefficient,and the test-re-test reliability was evaluated using the intraclass correlation coefficient(ICC).The construct validity of the Mandarin version of PRTEE scale was examined by analyzing correlation with the Chinese version of VAS and DASH scores using Pearson's correlation coefficient.The ceiling or floor effect was determined by explor-atory analysis.Result:The Cronbach's alpha coefficient of PRTEE total score,pain and function subscales were 0.947,0.872,and 0.927,respectively.The ICC values for test-retest reliability of PRTEE total score,pain,and func-tion subscales were 0.988、0.975,and 0.980,respectively.The correlation coefficient between PRTEE total score and its two subscales and VAS ranged from 0.838 to 0.920(P＜0.01),indicating a strong correlation.The correlation coefficient between the PRTEE total score and two subscales,the DASH total score and two di-mensions ranged from 0.704 to 0.893(P＜0.01),indicating a strong correlation.There were no subjects who re-ceived the lowest or highest scores,indicating that the scale had no ceiling or floor effects.Conclusion:The Mandarin version of PRTEE demonstrates good reliability and validity,with no evidence of ceiling or floor effect.It is suitable for evaluating tennis elbow in Mandarin-speaking patients.

Objective:This study analyzes the clinicopathological features and prognostic factors of early-stage gastric mixed adenoneuroendocrine carcinoma (G-MANEC), which is an exceedingly rare malignancy, in an effort to provide evidence-based insights for clinical decision-making.Methods:A retrospective observational study was conducted using the clinical data of 36 patients with early-stage G-MANEC who underwent surgical resection at Zhongshan Hospital, Fudan University, from July 2014 to May 2022. The observed indicators included clinicopathological data and follow-up information on recurrence, metastasis, and overall survival (OS).Results:Among the 36 patients there were 21 males and 15 females, aged 32-84 (65±11) years. The most common initial symptoms were abdominal pain and distension (19/36, 52.8%), followed by incidental findings during physical examinations (7/36, 19.4%). Tumors were located in the proximal stomach in 13 cases (36.1%), the middle stomach in 4 cases (11.1%), and the distal stomach in 19 cases (52.8%). Average tumor diameter was (2.48±1.18) cm. Gross morphology included elevated type in 12 cases (33.3%), flat type in 20 cases (55.6%), and depressed type in 4 cases (11.1%). Ulceration was present in 12 cases (33.3%). There were 11 cases (30.6%) at T1a stage and 25 cases (69.4%) at T1b stage. Lymph node metastasis was positive in 10 cases (27.8%), and the differentiation grades of the adenocarcinoma component were Grade I, II, and III in 3 (8.3%), 10 (27.8%), and 23 (63.9%) cases, respectively. Furthermore, the proportion of neuroendocrine carcinoma component was ≥50% in 18 cases (50.0%) and <50% in 18 cases (50.0%). Lymphovascular or perineural invasion was present in 18 cases (50.0%). Lauren classification included mixed type in 10 cases (27.8%), intestinal type in 19 cases (52.8%), and diffuse type in 7 cases (19.4%), and chromogranin A (CgA) positivity was found in 20 cases (55.6%). Additionally, the Ki-67 index positivity was found in 26 cases (72.2%). Total gastrectomy was performed in 12 cases (33.3%) and partial gastrectomy in 24 cases (66.7%), with a median follow-up duration of 77.5 months. The 3-year and 5-year OS rates were 88.89% and 79.67%, respectively. Univariate analysis revealed that age, gross morphology, ulceration, proportion of neuroendocrine carcinoma component, lymphovascular or perineural invasion, and chromogranin A (CgA) positivity showed statistical significance in their association with OS ( P<0.10). Multivariate Cox regression analysis further identified ulceration (HR=7.74, 95%CI: 1.24-48.30, P=0.028) and CgA positivity (HR=21.76, 95%CI: 1.86-53.97, P=0.014) as independent risk factors of OS. Conclusions:Patients with early-stage G-MANEC are typically asymptomatic, and those with ulceration or positive CgA immunohistochemical staining tend to have a poor prognosis.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective To investigate the effect of preoperative oral midazolam on postoperative de-lirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.Methods Eighty elderly patients undergoing laparoscopic surgery for radical resection of colorectal cancer,32 males and 48 females,aged 65-79 years,BMI 21-27 kg/m2,ASA physical status Ⅱ or Ⅲ,the state-trait anxiety inventory(STAI-S)≥38 scores at admission were selected.Patients were divided into two groups using random number method:control group and midazolam group,40 patients in each group.The midazolam group were administrated midazolam 7.5 mg per night till one day before surgery,while the placebo was administrated in the control group.The incidence of delirium 3 days after surgery and the STAI-S scores of one day before surgery were evaluated.The HR and MAP at entry,30 minutes after an-esthesia induction,1 hour,2 hours after anesthesia induction,and 30 minutes after extubation were recor-ded.The total dose of propofol,remifentanil and dexmedetomidine and the using rate of metaraminol were recorded.The visual analog scale scores 30 minutes after extubation,24 and 72 hours after surgery,the u-sing rate of tramadol,and the extubation time were recorded.Results Compared with the control group,the STAI-S scores of one day before surgery,and the incidence of postoperative delirium,the rate of using metaraminol,the VAS scores 30 minutes after extubation and 24 hours after surgery,the rate using of tram-adol were significantly decreased in the midazolam group(P＜0.05).There were no significant differences in total dose of propofol,remifentanil,and dexmedetomidine,extubation time between the two groups.Conclusion Preoperative oral midazolam can effectively reduce the incidence of postoperative delirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.

[Purpose]To analyze the trend of incidence and mean age at diagnosis of lymphoma in cancer registration areas of Jiangsu Province from 2009 to 2019.[Methods]Continuous and com-pleted data of cancer incidence from 2009 to 2019 were collected from 16 cancer registries in Jiangsu Province,and all indicators meeting the requirements.Joinpoint 4.7.0.0 software was used to analyze the trend of lymphoma incidence,the average annual percentage change(AAPC)and 95％confidence interval(CI)were calculated.A birth cohort was built from 1929 to 2019,to ana-lyze the change trend of lymphoma incidence rate among people born in different periods.[Re-sults]From 2009 to 2019,the AAPC of lymphoma incidence rate in cancer registration areas of Jiangsu Province was 5.74％,5.55％for men and 5.02％for women.The AAPC in urban areas was higher than that in rural areas(6.26％vs 4.90％).The age-standardized incidence rate(ASIR)of lymphoma increased by 3.40％annually on average.The rising amplitude of ASIR of women(3.16％)was higher than that of men(3.05％),and the rising amplitude of ASIR of lymphoma in urban areas(3.39％)was higher than that in rural areas(2.43％).The birth cohort analysis showed that the incidence rate of lymphoma fluctuated greatly before the age of 40,and there was no sig-nificant trend of change,while the incidence rate in age groups of 40 years old and above showed an overall upward trend.The average age at onset of lymphoma showed an upward trend from 2009 to 2019,with an average annual increase of 0.35 years old(P＜0.001).The standardized average age at onset for lymphoma still showed a significant upward trend,with an average annual increase of 0.15 years old(P=0.016).Compared with 2009,the standardized age specific incidence of lym-phoma in 2019 showed an overall backward trend,while the standardized incidence in all age groups after 60 years old(except for the age group of 70～74 years old)increased.The proportion of lymphoma in people above 60 years old was 56.16％in 2019,which was higher than that in 2009(52.57％).[Conclusion]The incidence rate of lymphoma in Jiangsu Province was increasing year by year from 2009 to 2019.The age at onset tended to shift back,and the trend was more pronounced in male than female.