Objective:To explore the clinicopathological and molecular genetic characteristics of anaplastic lymphoma kinase (ALK)-rearranged renal cell carcinoma (RCC), including a rare case with the TPM1-ALK gene subtype.Methods:Three cases of ALK-rearranged RCC diagnosed in the Department of Pathology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China from January 2020 to December 2024 were collected. Their clinical pathological and next-generation sequencing (NGS) data were analyzed. Relevant literature was also reviewed, and follow-ups were carried out.Results:Among the three patients, there were 1 female (case 1) and 2 males (cases 2 and 3), with the ages of 29,41 and 44 years, respectively. All of them were presented with space-occupying renal lesions. Case 1 (KIAA1217-ALK RCC) showed mixed cystic and solid components under the microscope, with tubular, papillary, and cribriform arrangements. The tumor cells had clear boundaries, and were cubic or low columnar, arranged in a single layer, pseudostratified or in sheets. The cytoplasm was abundant and eosinophilic, and part of the cytoplasm was vacuolated, as if there was accumulation of mucoid substances. The tumor cell nuclei were oval with prominent nucleoli. A large amount of mucus and inflammatory cell infiltration were noted in the stroma. Case 2 (TPM1-ALK RCC) showed a papillary growth pattern, with small, slender papillae accompanied by branches. The cells were arranged in a single layer, and the cytoplasm was either eosinophilic or clear. Foamy cells were aggregated in the stroma, accompanied by psammoma body-like calcifications. Case 3 (EML4-ALK RCC) was characterized by papillary and tubulocystic structures. The cytoplasm was abundant and eosinophilic. The tumor cell nuclei were large, with prominent nucleoli. There was conspicuous infiltration of lymphocytes and neutrophils in the fibromuscular stroma. The tumor cells all expressed epithelial markers, PAX8, GATA3, P504s and FH. ALK (5A4) staining showed diffuse strong expression in the cytoplasm, while TFE-3 was positive (nuclear stain) only in case 1 and case 3. The fluorescence in situ hybridization showed that ALK gene rearrangement was present in all three cases, while TFE-3 gene rearrangement/mutation was not detectable in case 1 and case 3. NGS showed the KIAA1217::ALK fusion (the fusion site in the exon 11 of KIAA1217 and exon 18 of ALK) in case 1, the TPM1::ALK fusion (the exon 8 of TPM1 and exon 20 of ALK) in case 2, and the EML4::ALK fusion (the exon 2 region of EML4 and the exon 20 region of ALK) in case 3.Conclusions:ALK-rearranged RCC has unique molecular characteristics. Its histological morphology is easily confused with that of papillary RCC and TFE3-rearranged RCC. Both immunohistochemistry and gene rearrangement tests should be used to confirm the diagnosis.

Objective:The minimal ablative margin (MAM) after radiofrequency ablation (RFA) was evaluated based on magnetic resonance imaging (MRI) image registration to analyze its effect on the prognosis of patients with hepatocellular carcinoma (HCC).Methods:Clinical data of 120 patients with HCC undergoing RFA in the General Hospital of Ningxia Medical University from January 2017 to April 2022 were retrospectively analyzed, including 88 males and 32 females, aged (58.4±8.5) years. The enhanced MRI images of patients before and after treatment were imported into a 3D Slicer software to show the ablative margin, and patients were divided into two groups according to whether MAM exceeded the peritumor safety boundary of 5 mm: MAM<5 mm group ( n=75) and MAM≥5 mm group ( n=45). Clinical data were recorded such as gender, age, tumor length and location. Patients were followed up by outpatient review to record whether local tumour progression occurred. Kaplan-Meier method was used for survival analysis, and log-rank test was used for survival comparison. Cox regression analysis was performed to analyze the risk factors of local tumour progression after RFA in patients with HCC. Results:There were significant differen-ces in tumor volume, whether the tumor is located around the vessels, and the mode of RFA guidance between the two groups (all P<0.05). The cumulative local tumour progression-free survival rates at 6, 12 and 24 months after RFA were 100%, 100% and 98% in MAM ≥5 mm group, superior to those in MAM<5 mm group (92%, 84% and 69%, respectively, χ2=47.22, P<0.001). Multivariate Cox regression analysis showed that MAM<5 mm ( OR=9.992, 95% CI: 4.358-22.913), tumor diameter ≥2 cm ( OR=1.758, 95% CI: 1.025-3.015) and perivascular tumor ( OR=2.344, 95% CI: 1.379-3.985) were risk factors for local tumour progression after RFA in patients with HCC (all P<0.05). Conclusion:The MAM evaluated based on MRI image registration is an influential factor on prognosis of patients with HCC. Patients with MAM<5 mm suffer an increased risk of postoperative local tumour progression.

Objective:To construct a prediction model for local tumor progression (LTP) in patients with hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA) based on the radiomics features of enhanced MRI.Methods:Clinical data of 120 patients with HCC undergoing RFA in the General Hospital of Ningxia Medical University from June 2017 to June 2022 were retrospectively analyzed, including 90 males and 30 females, aged (58.2±8.2) years. The patients were divided into training set ( n=84) and validation set ( n=36) in a ratio of 7∶3. According to whether LTP occurred within 2 years after RFA, the patients in training set were divided into LTP positive group ( n=32) and LTP negative group ( n=52). Logistic regression analysis was performed to analyze the risk factors for LTP after RFA in patients with HCC in training set. In the advanced arterial phase of preoperative enhanced MRI, the region of interest of tumor and peritumoral 5 mm area were mapped, and the radiomics features were extracted. The maximum correlation-minimum redundancy algorithm, the minimum absolute value shrinkage and selection operator algorithm were used to screen the radiomics features closely related to LTP, and the radiomics score was established. A nomogram model was constructed by combining the radiomics score with clinical tumor characteristics. The predictive performance and clinical practical value of different models were compared by the area under the receiver operating characteristic curve, calibration curve, clinical decision curve analysis (DCA) and clinical impact curve (CIC). Results:Tumor located around the blood vessels ( OR=4.574, 95% CI: 1.454-14.393, P=0.009) and ablation margin <5 mm ( OR=5.724, 95% CI: 1.996-16.420, P=0.001) were independent risk factors for LTP in patients with HCC after RFA. Five higher-order radiomics features were extracted and screened, including three tumoral features (glrlm_ShortRunHighGrayLevelEmphasis, ngtdm_Complexity and glcm_Imc1) and two peritumoral features (firstorder_Mean and glszm_SmallAreaHighGrayLevelEmphasis). Delong test showed that the area under curve of the combined model was higher than that of the radiomics model ( Z=2.90, P=0.004) and the clinical tumor characteristic model ( Z=2.56, P=0.010). Calibration curves, DCA and CIC curves all show that the combined model had a better clinical net benefit. Conclusion:Combining the radiomics features extracted from enhanced MRI images with clinical tumor characteristics can effectively predict the risk of LTP in patients with HCC after RFA.

ObjectiveTo investigate a suspected outbreak of healthcare-associated infection with carbapenem-resistant Klebsiella pneumoniae (CRKP) in a geriatric emergency ward, and to provide references for the prevention and control of multidrug-resistant bacteria in a hospital in Shanghai. MethodsOn-site epidemiological investigation, combined with environmental hygiene monitoring and pulsed field gel electrophoresis (PFGE) molecular typing method, were adopted to investigate a suspected outbreak of CRKP infection in the geriatric emergency ward of a hospital from October to November 2022, aiming at finding out factors caused the outbreak before taking corresponding control measures. ResultsA total of 3 cases of healthcare-associated CRKP infection were identified, of which 2 cases were homologous to a previous case of community-associated CRKP infection. What’s more, the 2 cases lived in the same ward with the latter and with adjacent beds, but the third case was non-homologous to the community-associated infection case. A total of 46 samples were collected from the environmental surfaces and the hands of healthcare workers, of which 7 samples tested positive for CRKP and were identical to the strains from the 2 healthcare-associated infection cases and the 1 community-associated infection case, originating from the bedrails, bedside tables, surface of non-invasive ventilator, bed curtains and panels of monitoring equipment, with a detection rate of 15.22%. But none of the 11 samples from the hands of healthcare workers tested positive for CRKP. The outbreak was effectively controlled after taking specific prevention and control measures such as strengthening personnel management, intensifying environmental cleaning and disinfection and strictly enforcing hand hygiene among healthcare workers. Subsequently, no similar new cases were reported during the 14-day follow-up period. ConclusionIncomplete environmental cleaning and disinfection, as well as inadequate enforcement of hand hygiene among heatheare workers may have contributed to the suspected outbreak of CRKP in the geriatric emergency ward. Early warning and timely investigation of suspected outbreaks of multidrug-resistant bacteria are crucial for preventing and controlling such outbreaks in hospitals.

Objective:The minimal ablative margin (MAM) after radiofrequency ablation (RFA) was evaluated based on magnetic resonance imaging (MRI) image registration to analyze its effect on the prognosis of patients with hepatocellular carcinoma (HCC).Methods:Clinical data of 120 patients with HCC undergoing RFA in the General Hospital of Ningxia Medical University from January 2017 to April 2022 were retrospectively analyzed, including 88 males and 32 females, aged (58.4±8.5) years. The enhanced MRI images of patients before and after treatment were imported into a 3D Slicer software to show the ablative margin, and patients were divided into two groups according to whether MAM exceeded the peritumor safety boundary of 5 mm: MAM<5 mm group ( n=75) and MAM≥5 mm group ( n=45). Clinical data were recorded such as gender, age, tumor length and location. Patients were followed up by outpatient review to record whether local tumour progression occurred. Kaplan-Meier method was used for survival analysis, and log-rank test was used for survival comparison. Cox regression analysis was performed to analyze the risk factors of local tumour progression after RFA in patients with HCC. Results:There were significant differen-ces in tumor volume, whether the tumor is located around the vessels, and the mode of RFA guidance between the two groups (all P<0.05). The cumulative local tumour progression-free survival rates at 6, 12 and 24 months after RFA were 100%, 100% and 98% in MAM ≥5 mm group, superior to those in MAM<5 mm group (92%, 84% and 69%, respectively, χ2=47.22, P<0.001). Multivariate Cox regression analysis showed that MAM<5 mm ( OR=9.992, 95% CI: 4.358-22.913), tumor diameter ≥2 cm ( OR=1.758, 95% CI: 1.025-3.015) and perivascular tumor ( OR=2.344, 95% CI: 1.379-3.985) were risk factors for local tumour progression after RFA in patients with HCC (all P<0.05). Conclusion:The MAM evaluated based on MRI image registration is an influential factor on prognosis of patients with HCC. Patients with MAM<5 mm suffer an increased risk of postoperative local tumour progression.

Objective:To construct a prediction model for local tumor progression (LTP) in patients with hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA) based on the radiomics features of enhanced MRI.Methods:Clinical data of 120 patients with HCC undergoing RFA in the General Hospital of Ningxia Medical University from June 2017 to June 2022 were retrospectively analyzed, including 90 males and 30 females, aged (58.2±8.2) years. The patients were divided into training set ( n=84) and validation set ( n=36) in a ratio of 7∶3. According to whether LTP occurred within 2 years after RFA, the patients in training set were divided into LTP positive group ( n=32) and LTP negative group ( n=52). Logistic regression analysis was performed to analyze the risk factors for LTP after RFA in patients with HCC in training set. In the advanced arterial phase of preoperative enhanced MRI, the region of interest of tumor and peritumoral 5 mm area were mapped, and the radiomics features were extracted. The maximum correlation-minimum redundancy algorithm, the minimum absolute value shrinkage and selection operator algorithm were used to screen the radiomics features closely related to LTP, and the radiomics score was established. A nomogram model was constructed by combining the radiomics score with clinical tumor characteristics. The predictive performance and clinical practical value of different models were compared by the area under the receiver operating characteristic curve, calibration curve, clinical decision curve analysis (DCA) and clinical impact curve (CIC). Results:Tumor located around the blood vessels ( OR=4.574, 95% CI: 1.454-14.393, P=0.009) and ablation margin <5 mm ( OR=5.724, 95% CI: 1.996-16.420, P=0.001) were independent risk factors for LTP in patients with HCC after RFA. Five higher-order radiomics features were extracted and screened, including three tumoral features (glrlm_ShortRunHighGrayLevelEmphasis, ngtdm_Complexity and glcm_Imc1) and two peritumoral features (firstorder_Mean and glszm_SmallAreaHighGrayLevelEmphasis). Delong test showed that the area under curve of the combined model was higher than that of the radiomics model ( Z=2.90, P=0.004) and the clinical tumor characteristic model ( Z=2.56, P=0.010). Calibration curves, DCA and CIC curves all show that the combined model had a better clinical net benefit. Conclusion:Combining the radiomics features extracted from enhanced MRI images with clinical tumor characteristics can effectively predict the risk of LTP in patients with HCC after RFA.

Objective:To explore the clinicopathological and molecular genetic characteristics of anaplastic lymphoma kinase (ALK)-rearranged renal cell carcinoma (RCC), including a rare case with the TPM1-ALK gene subtype.Methods:Three cases of ALK-rearranged RCC diagnosed in the Department of Pathology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China from January 2020 to December 2024 were collected. Their clinical pathological and next-generation sequencing (NGS) data were analyzed. Relevant literature was also reviewed, and follow-ups were carried out.Results:Among the three patients, there were 1 female (case 1) and 2 males (cases 2 and 3), with the ages of 29,41 and 44 years, respectively. All of them were presented with space-occupying renal lesions. Case 1 (KIAA1217-ALK RCC) showed mixed cystic and solid components under the microscope, with tubular, papillary, and cribriform arrangements. The tumor cells had clear boundaries, and were cubic or low columnar, arranged in a single layer, pseudostratified or in sheets. The cytoplasm was abundant and eosinophilic, and part of the cytoplasm was vacuolated, as if there was accumulation of mucoid substances. The tumor cell nuclei were oval with prominent nucleoli. A large amount of mucus and inflammatory cell infiltration were noted in the stroma. Case 2 (TPM1-ALK RCC) showed a papillary growth pattern, with small, slender papillae accompanied by branches. The cells were arranged in a single layer, and the cytoplasm was either eosinophilic or clear. Foamy cells were aggregated in the stroma, accompanied by psammoma body-like calcifications. Case 3 (EML4-ALK RCC) was characterized by papillary and tubulocystic structures. The cytoplasm was abundant and eosinophilic. The tumor cell nuclei were large, with prominent nucleoli. There was conspicuous infiltration of lymphocytes and neutrophils in the fibromuscular stroma. The tumor cells all expressed epithelial markers, PAX8, GATA3, P504s and FH. ALK (5A4) staining showed diffuse strong expression in the cytoplasm, while TFE-3 was positive (nuclear stain) only in case 1 and case 3. The fluorescence in situ hybridization showed that ALK gene rearrangement was present in all three cases, while TFE-3 gene rearrangement/mutation was not detectable in case 1 and case 3. NGS showed the KIAA1217::ALK fusion (the fusion site in the exon 11 of KIAA1217 and exon 18 of ALK) in case 1, the TPM1::ALK fusion (the exon 8 of TPM1 and exon 20 of ALK) in case 2, and the EML4::ALK fusion (the exon 2 region of EML4 and the exon 20 region of ALK) in case 3.Conclusions:ALK-rearranged RCC has unique molecular characteristics. Its histological morphology is easily confused with that of papillary RCC and TFE3-rearranged RCC. Both immunohistochemistry and gene rearrangement tests should be used to confirm the diagnosis.

Objective:To investigate the expression of circular RNA hsa_circ_0046701 in glioma tissues and its effect on the proliferation,migration and invasion of glioma U251 cells.Methods:Tumor tissue specimens and clinical data of fifty-two glioma patients treated surgically in Putuo People's Hospital Affiliated to Tongji University between June 2022 and March 2023 were collected,and another 30 normal brain tissue specimens were collected as controls.The expression level of hsa_circ_0046701 in glioma tissues was detected by qPCR to analyze the relationship between the level and clinical characteristics of the patient.Kaplan-Meier method was used to analyze the relationship between hsa_circ_0046701 level and survival prognosis.Using RNA interference technology,circ_0046701 overexpression and empty vector,siRNA-circ_0046701 and negative control(si-NC)plasmid were transfected into glioma U251 cells,respectively.The experiment cells were divided into the circ 0046701 OE group,Vector group,si-circ_0046701 group and si-NC group.CCK-8 method and Transwell assay were applied to detect the proliferation,migration and invasion abilities of cells in each group.Western blotting was applied to detect the expressions of vimentin,Snail,E-cadherin and cyclin D1 proteins in cells of each group.Results:The expression level of hsa_circ_0046701 in glioma tissues was significantly higher than that in normal brain tissues(P<0.01).The percentage of patients with WHO glioma grading(grades Ⅲ～Ⅳ)in the hsa_circ_0046701 high-expression group was significantly higher than that of the hsa_circ_0046701 low expression group(P<0.01).The OS of patients in the high-expression group was significantly shorter than that of the low-expression group.Compared with those in the si-NC group,the proliferation ability of U251 cells in the si-circ_0046701 group was significantly reduced(P<0.05 or P<0.01),and the numbers of migrated and invaded cells decreased significantly(both P<0.01).The expressions of vimentin,Snail,and cyclin D1 proteins in the cells was significantly reduced(all P<0.01),while the expression of E-cadherin protein increased significantly(P<0.01).Compared with those in the Vector group,the proliferation ability of U251cells in the circ_0046701 OE group increased significantly(P<0.01);the numbers of migrated and invaded cells increased significantly(both P<0.01);the expressions of vimentin,Snail,and cyclin D1 proteins in the cells increaseds significantly(all P<0.01),and the expression of E-cadherin protein decreased significantly(P<0.01).Conclusion:Circular RNA hsa_circ_0046701 is highly expressed in glioma tissues,and is correlated with poor patient prognosis.Knockdown of hsa_circ_0046701 expression could inhibit the proliferation,migration and invasion of glioma U251cells.

BACKGROUND:Warm-needling moxibustion can effectively treat knee osteoarthritis.Degeneration,injury and fracture of the anterior cruciate ligament can affect the local stability of the knee joint,and then induce the formation of knee osteoarthritis.Whether warm-needling moxibustion can repair the injured cruciate ligament and the mechanism of action are still unclear. OBJECTIVE:To observe the effects of warm-needling moxibustion on the morphology of the anterior cruciate ligament and the expression of insulin growth factor-1 and transforming growth factor-β in rabbits with knee osteoarthritis and to clarify the mechanism of anterior cruciate ligament repair by warm-needling moxibustion. METHODS:Thirty New Zealand rabbits were randomly divided into blank group,model group and warm-needling moxibustion group,with 10 rabbits in each group.Knee osteoarthritis model was established by plaster cast immobilization.The blank group was not intervened.Rabbits in the model group rabbits were fixed in a rabbit holder for 15 minutes every day.The warm-needling moxibustion group was treated with warm acupuncture,once a day,7 days as a course of treatment,a total of two courses.After treatment,the imaging changes of the anterior cruciate ligament were observed by MRI and MRI grading statistics were performed.Morphological changes of the anterior cruciate ligament were observed by transmission electron microscope and hematoxylin-eosin staining.mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:MRI examination:Compared with the blank control group,the anterior cruciate ligament in the model group was thickened,edematous,and partially torn,and the difference in grading statistics was statistically significant(P＜0.05).Compared with the model group,the anterior cruciate ligament in the warm-needling moxibustion group was slightly thickened,with mild edema and no tearing,and the difference in grading statistics was statistically significant(P＜0.05).General observation:In the model group,the surface of the anterior cruciate ligament was glossy and faded,with the edge being covered with flocculent periosteum and obvious tissue necrosis;in the warm-needling moxibustion group,the surface of the ligament was glossy,and the ligament was in a normal helical shape.Hematoxylin-eosin staining:In the model group,there was obvious tissue necrosis in the anterior cruciate ligament,a large number of new capillaries,loosely arranged fibroblasts and collagen fibers.In the warm-needling moxibustion group,there was a small amount of tissue necrosis and few new vessels in the anterior cruciate ligament,and the cells and collagen fibers were loosely and irregularly arranged.Transmission electron microscopy:In the model group,the fibers in the anterior cruciate ligament were arranged in a disordered way with uneven thickness and distribution,and there are more fibroblasts that were irregular in morphology;in the warm acupuncture group,the fibers were basically arranged longitudinally,with uneven thickness and distribution,and a small number of oval-shaped fibroblasts were observed.RT-PCR and western blot assay:mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were significantly decreased in the model group compared with the blank control group(P＜0.05),but significant increased after treatment with warm-needling moxibustion(P＜0.05).To conclude,warm-needling moxibustion can alleviate anterior cruciate ligament injury and regulate the expression of insulin growth factor-1 and transforming growth factor-β to treat knee osteoarthritis.

Objective:To investigate the association between exposure to ambient air pollutants and pregnancy outcomes in patients undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI), and to clarify the susceptible windows of exposure. Methods:A retrospective cohort study was conducted on 2 071 infertile women aged ≤40 years who underwent the first fresh cleavage stage embryo transfer in the Department of Reproductive Medicine of the First Affiliated Hospital of Harbin Medical University from January 2014 to December 2021. Patients were divided into heating season group and non-heating season group based on whether the start date of gonadotropin (Gn) was during the heating season or not, and baseline data and pregnancy outcomes were compared between the two groups. Daily average levels of six criteria air pollutants [fine particulate matter (PM 2.5), inhalable particulate matter (PM 10), nitrogen dioxide (NO 2), sulfur dioxide (SO 2), carbon monoxide (CO), ozone (O 3) max-8 h] in four discrete periods were obtained. The four discrete periods included period 1, 75 d prior to Gn start to Gn start; period 2, Gn start to oocyte retrieval; period 3, oocyte retrieval to day 3 embryo transfer; period 4, embryo transfer to serum human chorionic gonadotropin test. The primary outcome was ongoing pregnancy. A multiple logistic regression model was used to investigate the association between air pollutant exposure and pregnancy outcomes, adjusted for important confounders. Results:The biochemical pregnancy rate [51.7% (465/900)], the clinical pregnancy rate [45.2% (407/900)], the ongoing pregnancy rate [38.2% (344/900)], and the live birth rate [36.1% (325/900)] in the heating season group were significantly lower than those in the non-heating season group [56.3% (659/1 171), P=0.037; 51.2% (599/1 171), P=0.007; 44.3% (519/1 171), P=0.005; 41.3% (484/1 171), P=0.016]. A significant negative correlation was observed between SO 2 and NO 2 exposure and ongoing pregnancy in all four time windows. SO 2 increased by one interquartile range (IQR), corresponding to adjusted OR (a OR) and 95% CI were 0.92 (0.85-0.99), 0.92 (0.87-0.99), 0.93 (0.87-0.99) and 0.93 (0.87-0.99), respectively. An IQR increase in NO 2 was also significantly associated with decreased odds of ongoing pregnancy (a OR=0.83, 95% CI: 0.72-0.95; a OR=0.87, 95% CI: 0.77-0.97; a OR=0.90, 95% CI: 0.81-1.00; a OR=0.85, 95% CI: 0.77-0.95, respectively). During period 1 and period 4, we observed adverse effects of PM 10 exposure on ongoing pregnancy (a OR=0.86, 95% CI: 0.75-0.98; a OR=0.89, 95% CI: 0.80-1.00, respectively). In Period 1, PM 2.5 exposure was significantly associated with reduced odds for ongoing pregnancy with a OR=0.86, 95% CI: 0.75-0.99. In addition, NO 2 exposure was associated with a decreased likelihood of achieving clinical pregnancy across all exposure windows except for period 3. However, no associations were noted with CO and O 3. Conclusion:Ambient air pollution has detrimental effects on pregnancy outcomes in patients undergoing fresh embryo transfer. Notably, the adverse impacts were also observed during preantral-antral follicle transition stage before IVF/ICSI treatment. A significant negative association between NO 2 exposure and pregnancy outcomes was observed in almost all exposure windows, indicating that NO 2 may be the main air pollutant associated with adverse pregnancy outcomes.