Polycystic ovary syndrome (PCOS) is the predominant cause of subfertility in reproductive-aged women; however, its pathophysiology remains unknown. Neurotensin (NTS) is a member of the gut-brain peptide family and is involved in ovulation; its relationship with PCOS is unclear. Here, we found that NTS expression in ovarian granulosa cells and follicular fluids was markedly decreased in patients with PCOS. In the in vitro culture of cumulus-oocyte complexes, the neurotensin receptor 1 (NTSR1) antagonist SR48692 blocked cumulus expansion and oocyte meiotic maturation by inhibiting metabolic cooperation and damaging the mitochondrial structure in oocytes and surrounding cumulus cells. Furthermore, the ERK1/2-early growth response 1 pathway was found to be a key downstream mediator of NTS/NTSR1 in the ovulatory process. Animal studies showed that in vivo injection of SR48692 in mice reduced ovulation efficiency and contributed to irregular estrus cycles and polycystic ovary morphology. By contrast, NTS partially ameliorated the ovarian abnormalities in mice with dehydroepiandrosterone-induced PCOS. Our findings highlighted the critical role of NTS reduction and consequent abnormal NTSR1 signaling in the ovulatory dysfunction of PCOS, suggesting a potential strategy for PCOS treatment.
Polycystic Ovary Syndrome/physiopathology* ; Female ; Animals ; Neurotensin/metabolism* ; Receptors, Neurotensin/antagonists & inhibitors* ; Mice ; Ovulation/drug effects* ; Humans ; Granulosa Cells/metabolism* ; Adult ; Oocytes/metabolism* ; MAP Kinase Signaling System ; Signal Transduction ; Follicular Fluid/metabolism* ; Disease Models, Animal ; Gonadotropin-Releasing Hormone/analogs & derivatives*

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

Objective To investigate the effects of miR-338-3p on interleukin(IL)-13-induced human bronchial epithelial cell line(BEAS-2B)injury and airway inflammation in mice with ovalbumin(OVA)-induced asthma.Methods OVA was used to replicate an asthma model of mice,which were divided into control group,model group,miR-NC agomir and miR-338-3p agomir intervention groups.HE staining microscopy was employed to ob-serve the pathological morphology of lung tissue,while TUNEL staining was used to assess cell apoptosis in lung tis-sue.ELISA was conducted to measure the levels of interleukin-1β(IL-1β)and tumor necrosis factor(TNF)-α in lung tissue.The BEAS-2B cells were subjected to IL-13-induced injury and divided into control group,IL-13 group,IL-13+miR-NC group,and IL-13+miR-338-3p mimic group.Cell viability was assessed with MTT assay.Flow cytometry was employed to evaluate cell apoptosis.The level of IL-1β and TNF-α in cells was measured by ELISA.The targeting relationship between miR-338-3p and Ras homologous(Rho)was investigated using bioinfor-matics analysis,luciferase assay,Western blot,and functional repair assay.Results Compared to the model group,the miR-338-3p agamid intervention group exhibited a significant reduction in inflammatory cell infiltration and airway wall thickening in lung tissue,as well as decreased cell apoptosis and the level of IL-1β and TNF-α in lung tissue(P＜0.05).Compared to the control group,cell viability of BEAS-2B cells in the IL-13+miR-338-3p mimic group exhibited a significant increase(P＜0.05),while apoptosis and level of IL-1β and TNF-α within the cells demonstrated a notable decrease(P＜0.05).Rho was a target gene of miR-338-3p,and over-expression of Rho attenuated the effect of miR-338-3p mimic on IL-13-induced injury and inflammation in BEAS-2B cells.Conclusions Up-regulation of miR-338-3p can inhibit asthma-related airway inflammation and injury of lung epi-thelial cells with a potential mechanism targeting at Rho gene.

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

Objective:To explore the application value and treatment effects of multiple free anterolateral perforator flaps of calf for reconstruction of multiple digital wounds in single hand.Methods:From August 2020 to March 2022, 12 patients with soft tissue defects in 35 digits were treated in Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. Ten patients were male and 2 were female, aged 25 to 58 years old. Of the patients, 1 had soft tissue defects in 5 digits, 3 in 4 digits, 2 in 3 digits and 6 in 2 digits. The size of defects was from 1.2 cm ×1.2 cm to 7.0 cm×3.5 cm after debridement. The vascular perforators discovered from intraoperative explorations were found originating from the superficial peroneal artery in 24 flaps, from the peroneal artery in 7 flaps and from the anterior tibial artery in 4 flaps. During surgery, the perforator artery and accompanying veins of the flaps were anastomosed with the proper digital artery and palmar or dorsal subcutaneous veins in the recipient site, respectively. The size of the flaps was from 1.5 cm×1.5 cm to 7.5 cm×4.0 cm. No nerve was affected in the surgery. The wound at donor sites in the calf was sutured directly. Regular postoperative follow-ups were conducted at outpatient clinics. The comprehensive evaluation scale of flap was used to assess the conditions of the donor and recipient sites.Results:In this study, all 35 soft tissue defects of digits in 12 patients were reconstructed by the anterolateral perforator flaps of calf. All the 35 flaps survived after surgery, with a 100% of survival rate. The patients were instructed to carry out early functional training after surgery. Follow-up lasted 6 to 24 months, with an average of 11 months. Twenty-five flaps were found in slightly swollen, and further flap thinning surgery were carried out 3 months after the primary surgery, while the rest of the flaps had good appearance and texture. At 6 months after surgery, all flaps recovered a partial deep and shallow sensory and sense of touch. All wound at donor sites in calf had one-stage healing without dysfunction. The comprehensive evaluation scale was excellent in 28 flaps and good in 7 flaps. The excellent and good rate was 100%.Conclusion:It is an effective method to use multiple free anterolateral perforator flaps of calf to reconstruct multiple digit defects in single hand. The flaps can be conveniently harvested and the multiple digital defects in single hand can be reconstructed in primary surgery with small damages to the donor sites and together with satisfactory clinical outcomes.

Objective To observe the clinical and CT features of neonatal adrenal cystic neuroblastoma(CNB).Methods Eight newborns with adrenal CNB confirmed by surgical pathology were retrospectively analyzed.The clinical data were recorded,and the plain and enhancement abdominal CT manifestations were observed.Results Among 8 cases,6(6/8,75.00％)were detected with prenatal ultrasound,while 2(2/8,25.00％)were detected after birth with ultrasound,all with single adrenal grand lesion,located half in left and half in right adrenal gland(each 4/8,50.00％).The maximum diameter of CNB lesion was 2.3-6.1 cm,with the median maximum diameter of 4.5 cm.CT showed all 8 lesions(8/8,100％)presented as single localized adrenal grand thick-walled cystic lesion,among which 3(3/8,37.50％)with uniform density within the cysts,3(3/8,37.50％)with internal septum within the cysts,1(1/8,12.50％)with slight floating debris and the rest 1(1/8,12.50％)with both internal septum and floating debris in the cyst.No calcification,cross the midline nor surround blood vessels were observed.Seven(7/8,87.50％)lesions had clear while 1(1/8,12.50％)had unclear boundaries,all mildly compressed surrounding structures.The capsule wall and internal septum of the cysts slightly enhanced after enhancement.Multiple liver metastases occurred in 2 cases.Conclusion Most neonatal adrenal CNB were detected before delivery,which mainly presented as thick-walled cystic mass with clear boundary,accompanied by septa and floating debris,and the cystic wall and septa slightly enhanced after enhancement,and liver metastasis might occur.

Objective:To investigate the value of CT-radiomics based machine learning model in predicting the abundance of tumor infiltrating CD8 +T cells and the prognosis of pancreatic cancer patients. Methods:A total of 150 pancreatic cancer patients who underwent surgical excision and confirmed by pathology from Fudan University Shanghai Cancer Center between December 2011 and January 2017 were retrospectively enrolled. The patients were randomly divided into the training set ( n=105) and the validation set ( n=45) in a 7∶3 ratio with simple random sampling. The immunohistochemical method was used to assess the abundance of tumor infiltrating CD8 +T cells, and the patients were then divided into high infiltrating group ( n=75) and low infiltrating group ( n=75) according to the median. The prognosis between the 2 groups was evaluated using Kaplan-Meier method and log-rank test. Radiomic features were extracted from preoperative venous-phase enhanced CT images in the training set. The Wilcoxon test, the max-relevance and min-redundancy algorithm were used to select the optimal feature set. Three supervised machine learning models (decision tree, random forest and extra tree) were established based on the optimal feature set to predict the abundance of tumor infiltrating CD8 +T cells. Performance of above-mentioned models to predict the abundance of tumor infiltrating CD8 +T cells in pancreatic cancer was tested in the validation set. The evaluation parameters included area under the receiver operating characteristic curve (AUC), F1-score, accuracy, precision and recall. Results:The median overall survival time of patients in high infiltrating group and low infiltrating group were 875 days and 529 days, respectively (χ2=11.53, P<0.001). The optimal feature set consisted of 10 radiomic features in training set. In the validation set, the decision tree, random forest and extra tree model showed the AUC of 0.620, 0.704 and 0.745, respectively; corresponding to a F1-score of 0.457, 0.667 and 0.744, the accuracy of 57.8%, 68.9% and 75.6%, the precision of 66.7%, 73.7% and 80.0%, the recall of 34.8%, 60.9% and 69.6%. Conclusions:Pancreatic cancer patients with high tumor infiltrating CD8 +T cells have better prognosis than those with low tumor infiltrating CD8 +T cells. The radiomics-based extra tree model is valuable in predicting the CD8 +T cells infiltrating level in pancreatic cancer.

ObjectiveWe examined the principal respiratory pathogens in patients with acute respiratory tract infection in Taizhou， Zhejiang Province during 2020‒2021 to provide evidence for prevention， diagnosis and treatment of acute respiratory tract infection. MethodsFrom September 2020 to August 2021， a total of 2 831 cases with acute respiratory tract infection were collected from two influenza sentinel surveillance hospitals in Taizhou， which had then received the examination of 22 respiratory pathogens by multiple fluorescence quantitative PCR. ResultsThe total positive rate of respiratory pathogens in 2 831 samples was 14.13%， among which enterovirus （7.77%） and respiratory syncytial virus （1.59%） were the principal pathogens. Except enterovirus， there was no significant difference in the positive rate of pathogens detected by gender（P>0.05）. Moreover， there was significant difference in pathogens by age （P<0.05）， with the highest positive rate in 0‒4 years（35.21%）. There was also significant difference in pathogens by seasons （P<0.05）， with the highest positive rate in summer（20.54%）. ConclusionThe positive rate of acute respiratory tract infection decreases significantly， compared with that before the COVID-19 epidemic. The differences in the positive rate differ significantly by age and seasons. Comprehensive consideration of diverse factors before diagnosis and the utilization of multiple fluorescent quantitative PCR can quickly and effectively determine the pathogens in the early stage of infection. Our findings may provide certain support for the diagnosis and treatment of acute respiratory infections in the context of COVID-19 in Taizhou.

Objective:To investigate the effect of CRT on thapsigargin (TG)-induced epithelial mesenchymal transition (EMT) of pancreatic cancer (PC) cells.Methods:Immunohistochemistry was used to investigate the differential CRT expression in PC tissues. Western blot (WB) and transwell were used to detect the effect of CRT silencing on TG induced EMT phenotype. Fluro-4/AM and confocal microscopy were used to detect intracellular calcium level in PC cells.Results:CRT was overexpressed in PC tissues ( P<0.01). Overexpression of CRT was positively associated with lymph node metastasis ( P=0.017) and UICC stage ( P=0.021) of PC patients, and negatively associated with E-cadherin expression ( P=0.013). High CRT and low E-cad expression contributed to the poor prognosis of PC patients ( P=0.023). In PC cells, TG induced EMT phenotype was reversed by siRNA-mediated CRT silencing. TG induced EMT was significantly reversed by CRT silencing in vitro. Conclusions:CRT mediates TG induced intracytoplasmic Ca 2+, and ultimately promotes EMT of PC cells.

Objective:To investigate the effects of anterolateral wide pedicled double dynamic flap of the calf in repair of soft tissue defects of mid-and forefoot.Methods:From September 2015 to Septemler 2020, 15 cases with severe soft tissue defects of mid-and forefoot were repaired with the anterolateral wide pedicled double dynamic flap of the calf. There were 11 males and 4 females with an average age of 37(range, 22-53)years old. Of the 15 cases, the defects were caused by traffic accident in 6 cases and objects smash in 9 cases. Three cases were simple soft tissue defect, and 12 cases combined with fracture or dislocation and bone defect. The size of soft tissue defects ranged from 4 cm×5 cm to 7 cm×12 cm. All wounds of donor sites were repaired by skin grafting. All patients entered follow-ups at the outpatient clinic or through WeChat. The appearance of flaps and limb recovery were recorded after surgery.Results:All cases followed-up for 6-24 (mean, 16) months. Two days after surgery, 1 case had flap swelling and cyanosis, which was improved after pedicle suture removal and surface bloodletting. The pedicle of the flap was slightly bloated in 4 cases, and the texture and appearance were good in 11 cases. The ankle function of all cases recovered satisfactorily. The ranges of ankle motion were 15°-20° for dorsiflexion and 30°-40° for plantar flexion. The donor site healed well and all the skin grafts survived.Conclusion:The anterolateral wide pedicled double dynamic flap of the calf is one of the ideal flaps for repairing the soft tissue defects of the mid-and forefoot with reliable blood supply, sufficient venous return, simple operation and no require a vascular anastomosis.