Objective: To evaluate the incidence, treatment, and survival outcomes of Swyer syndrome with gonadal non-dysgerminoma malignant germ cell tumor (MGCT-NDG). Methods: A retrospective study was performed on Swyer syndrome patients with MGCT-NDG between January 2011 and December 2022 in Peking Union Medical College Hospital to investigate their characteristics and outcomes. Results: A total of 15 patients (4.9%, 15/307) with Swyer syndrome were identified in 307 MGCT-NDG patients. The average age at diagnosis of MGCT-NDG and Swyer syndrome were (16.8±6.7) and (16.7±6.6) years, respectively. Six cases were preoperatively diagnosed as Swyer syndrome, of which 4 cases received bilateral gonadectomy with or without hysterectomy, while the other 2 cases underwent removal of gonadal tumor and unilateral gonadectomy with hysterectomy, respectively. Of the 9 patients postoperatively diagnosed as Swyer syndrome, unilateral gonadectomy, removal of gonadal tumor, and unilateral gonadectomy with hysterectomy were performed in 6 patients, 2 patients, and 1 patient, respectively. Mixed malignant germ cell tumor (MGCT;10 cases), yolk sac tumor (4 cases), and immature teratoma (1 case) were the pathological subtypes, in the descending order. There were International Federation of Gynecology and Obstetrics (FIGO) stage Ⅰ in 6 cases, stage Ⅱ in 3 cases, stage Ⅲ in 5 cases, and stage Ⅳ in 1 case, respectively. Eleven patients received reoperation for residual gonadectomy after a average delay of (7.9±6.2) months, including 8 MGCT-NDG patients and 1 gonadoblastoma patient, no tumor involved was seen in the remaining gonads in the other 2 cases. Ten patients experienced at least one recurrence, with a median event free survival of 9 months (5, 30 months), of which 2 patients received surgery only at the time of initial treatment. All patients with recurrence received surgery and combined with postoperative chemotherapy. After a median follow-up of 25 months (15, 42 months), 10 patients were disease-free, 3 patients died of the tumor, 1 died of side effects of leukemia chemotherapy, and 1 survived with disease. Conclusion: The incidence rate of Swyer syndrome in patients with MGCT-NDG is about 4.9%; timely diagnosis and bilateral gonadectomy should be emphasized to reduce the risk of reoperation and second carcinogenesis in this population.
Female ; Humans ; Retrospective Studies ; Gonadal Dysgenesis, 46,XY/surgery* ; Gonadoblastoma/surgery* ; Neoplasms, Germ Cell and Embryonal/surgery* ; Ovarian Neoplasms/pathology*

OBJECTIVE To explore the molecular mechanism of urolithin A inhibition of human hepatoma cells growth. METHODS Hepatoma Huh-7 cells were treated with different concentrations of urolithin A(Uro-A). The inhibition rate of Huh-7 cells survival was detected by CCK-8 assay and the IC50 was calculated. Cell proliferation was detected by colony formation assay and cell migration ability was assessed by cell wound healing experiment. Glucose uptake and lactate level in culture medium through colorimetry and the ATP production in cell through chemiluminescence method was analyzed. Western blotting was applied to detect protein expression levels of glucose transporter(GLUT1), key enzymes of glycolysis(HK2, PFKM, LDHA), p53, p-p38 and Bcl-2 after treatment with different concentrations of Uro-A. Flow cytometry and TUNEL method were used to detect apoptosis rate. RESULTS The results of CCK-8 showed that Uro-A significantly inhibited the proliferation of Huh-7 cells, and the IC50 was(48.54±1.21) μmol·L−1. The ability of clone formation and migration decreased after Uro-A treatment. Cellular glucose uptake and level of lactic acid and ATP production were down regulated in Huh-7 cells treated with Uro-A. The results showed that expression of glycolytic key proteins GLUT1, PKM2, LDHA and HK2 decreased. Western Blotting further research indicated that the p53 and p-p38 were activated, while the Bcl-2 was down-regulated. Flow cytometry data and TUNEL method revealed that the induction of apoptosis by Uro-A was remarkably increased. CONCLUSION These findings suggest that Uro-A can suppress Huh-7 cell proliferation and migration. The possible mechanism is the inhibition of glycolysis by p53, p-p38 and Bcl-2, which prevent cell growth and finally induce apoptosis.

OBJECTIVES: To study the effect of ligustrazine injection on mitophagy in neonatal rats with hypoxic-ischemic encephalopathy (HIE) and its molecular mechanism. METHODS: Neonatal Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group with 8 rats, a model group with 12 rats, and a ligustrazine group with 12 rats. The rats in the model group and the ligustrazine group were used to establish a neonatal rat model of HIE by ligation of the left common carotid artery followed by hypoxia treatment, and blood vessels were exposed without any other treatment for the rats in the sham-operation group. The rats in the ligustrazine group were intraperitoneally injected with ligustrazine (20 mg/kg) daily after hypoxia-ischemia, and those in the sham-operation group and the model group were intraperitoneally injected with an equal volume of normal saline daily. Samples were collected after 7 days of treatment. Hematoxylin and eosin staining and Nissl staining were used to observe the pathological changes of neurons in brain tissue; immunohistochemical staining was used to observe the positive expression of PINK1 and Parkin in the hippocampus and cortex; TUNEL staining was used to measure neuronal apoptosis; Western blotting was used to measure the expression levels of the mitophagy pathway proteins PINK1 and Parkin and the autophagy-related proteins Beclin-1, microtubule-associated protein 1 light chain 3 (LC3), and ubiquitin-binding protein (P62). RESULTS: Compared with the sham-operation group, the model group had a significant reduction in the number of neurons, an increase in intercellular space, loose arrangement, lipid vacuolization, and a reduction in Nissl bodies. The increased positive expression of PINK1 and Parkin, apoptosis rate of neurons, and protein expression levels of PINK1, Parkin, Beclin1 and LC3 (P<0.05) and the decreased protein expression level of P62 in the hippocampus were also observed in the model group (P<0.05). Compared with the model group, the ligustrazine group had a significant increase in the number of neurons with ordered arrangement and an increase in Nissl bodies, significant reductions in the positive expression of PINK1 and Parkin, the apoptosis rate of neurons, and the protein expression levels of PINK1, Parkin, Beclin1, and LC3 (P<0.05), and a significant increase in the protein expression level of P62 (P<0.05). CONCLUSIONS Ligustrazine can alleviate hypoxic-ischemic brain damage and inhibit neuronal apoptosis in neonatal rats to a certain extent, possibly by inhibiting PINK1/Parkin-mediated autophagy.
Rats ; Animals ; Hypoxia-Ischemia, Brain/metabolism* ; Animals, Newborn ; Rats, Sprague-Dawley ; Beclin-1 ; Autophagy ; Ubiquitin-Protein Ligases/metabolism* ; Protein Kinases/metabolism*

【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Polysaccharide of Balanophora involucrata Hook. f. (BPS), the major component of Balanophora involucrata Hook. f., was confirmed the protective effect on liver injury in our previous study. This research aimed to investigate the protective mechanism of BPS on experimental liver injury by attenuating cell ferroptosis through modulating solute carrier family 7 member 11/glutathione peroxidase 4 (SLC7A11/GPX4) pathway. The animal experiment was approved by the Experimental Animal Ethical Committee of Hubei Minzu University and all rats had received human care in compliance with the institutional animal care guidelines. Rats were given intraperitoneal injection of (D-galactosamine, D-GalN) solution (800 mg·kg^-1) one time to establish the acute liver injury model. The results showed aspartate amino transferase (AST), alanine aminotransferase (ALT) and 4-hydroxynonenal (4-HNE) levels in serum were decreased, and the contents of reactive oxygen species (ROS), Fe²⁺, malondialdehyde (MDA) and lipid peroxide (LPO) in liver tissues also decreased and glutathione (GSH) level increased after BPS administration with 200 mg·kg^-1. Besides, BPS reduced iron deposition and increased the expression of SLC7A11 and GPX4 proteins in liver tissue. In conclusion, BPS ameliorated experimental liver injury by alleviating cell ferroptosis through SLC7A11/GPX4 pathway. The present study pointed to the possibility of utilizing BPS for protection against liver injury in clinic.

ObjectiveTo observe the effect of Shenghuitang on serum levels of neurotransmitters in the mouse model of Alzheimer's disease （AD） and explore the mechanism of Shenghuitang in mitigating the cognitive impairment and circadian rhythm disturbance of AD. MethodTwenty-seven APP/PS1 dementia mice were randomly assigned into a model group， a donepezil （0.92×10^-4 g·kg^-1·d^-1） group， and a Shenghuitang （13.5 g·kg^-1·d^-1） group. Another nine wild-type C57BL/6JNju mice was set as the control group. The mice were administrated with corresponding drugs by gavage and the control and model groups were given the same volume of pure water. Every group was continuously treated for 4 weeks. The cognitive function and circadian rhythm of mice were evaluated by Morris water maze test and open field test. The liquid chromatography-tandem mass spectrometry （LC-MS/MS） was employed to determine the expression levels of acetylcholine （ACh）， choline acetyltransferase （ChAT）， norepinephrine （NE）， epinephrine （E）， glutamate （Glu）， 5-hydroxyindoleacetic acid （5-HIAA）， and dopamine （DA） in the serum. ResultCompared with the control group， the modeling increased the escape latency， swimming distance， time of first arrival on the platform， activity time of light environment， activity time of dark environment， and total activity time （P<0.01）， while it decreased the number of crossing the platform and the swimming time in the target quadrant （P<0.01）. Compared with the model group， donepezil and Shenghuitang decreased the escape latency， swimming distance， time of first arrival on the platform， activity time of light environment， activity time of dark environment and total activity time （P<0.05， P<0.01）， while they increased the number of crossing the platform and the swimming time in the target quadrant （P<0.05， P<0.01）. Compared with the control group， the modeling down-regulated the expression levels of ACh and ChAT in the serum （P<0.01）. Compared with the model group， donepezil and Shenghuitang up-regulated the expression levels of ACh and ChAT in the serum （P<0.05， P<0.01）. Compared with the control group， the modeling up-regulated the expression level of Glu in the serum （P<0.01） and down-regulated the expression levels of NE， 5-HIAA， and DA （P<0.01）. Compared with the model group， Shenghuitang down-regulated the expression level of Glu （P<0.05） and up-regulated the expression levels of NE， 5-HIAA， and DA （P<0.05， P<0.01）. The expression levels of NE， Glu， 5-HIAA， and DA in the donepezil group did not change significantly compared with those in the model group. The expression level of E showed no significant difference among different groups. ConclusionShenghuitang may ameliorate the cognitive impairment and circadian rhythm disturbance of AD mice by regulating the levels of neurotransmitters in the serum.

Exosomes have the dual characteristics of promoting and inhibiting cancer and can induce the proliferation, invasion, and metastasis of hepatoma cells by altering tumor microenvironment, promoting neovascularization, and regulating epithelial-mesenchymal transition. Exosomes can regulate hepatocellular carcinoma and inhibit the growth and metastasis of hepatoma cells by regulating a variety of physiological and pathological processes, thus playing an important role in clinical diagnosis and treatment. It is pointed out that exosomes may become an effective antitumor treatment method through immune regulation and remodeling of tumor microcirculation.

[Abstract] Objective: To investigate the effect of polyadenosine diphosphate ribose polymerase 1 (PARP1) on the proliferation and 5-FU resistance of gastric cancer cells and its potential mechanism. Methods: The tumor tissues and corresponding paracancerous tissues of 72 patients with gastric cancer who were treated in Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from May 2018 to December 2019 were collected. AGS cells were transfected with siFUT8 to knock down FUT8 gene expression. qPCR and immunohistochemistry were used to detect the expression of PARP1 in gastric cancer and adjacent tissues. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis and colony formation of AGS cells. MTT assay was used to detect the effect of AG14361 on the 5-FU sensitivity of gastric cancer cells. The overall distribution of differential genes in AGS cells treated with AG14361 was analyzed by mRNA sequencing, and related signaling pathways were analyzed by KEGG enrichment. qPCR and WB were used to detect the expression of α-1,6-fucosyltransferase (FUT8) in AGS cells and the interference effect of FUT8. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis, and colony formation of AGS cells disturbed by siFUT8. Results: Compared with paracancer tissues, PARP1 expression was significantly increased in gastric cancer tissues (P<0.001). AG14361 can significantly inhibit the proliferation and colony formation of AGS cells, thus promoting the apoptosis of AGS cells (all P<0.01). AG14361 treatment reduced the IC50 of 5-FU against gastric cancer cells, especially against AGS cells, with IC50 decreased by more than 60%. mRNA sequencing results showed that FUT8 was a key glycosyltransferase of AG14361 in inhibiting the proliferation of AGS cells (P<0.05). Compared with the siNC group, treatment of AG14361 with IC50 significantly reversed the promotion of AGS cells proliferation caused by inerference with FUT8, promoted apoptosis and BAX protein expression, decreased Bcl2 protein expression and inhibited the increase in AGS cell colony formation caused by interference with FUT8 (all P<0.01). Conclusion: PARP1 can promote malignant transformation of gastric cancer cells by regulating N-glycosyltransferase FUT8. AG14361 can enhance the chemotherapy sensitivity of 5-FU, and PARP1 may become a potential target for gastric cancer treatment.

This study was designed to identify the pathogen causing soft rot of Pinellia ternata in Qianjiang of Hubei province and screen out the effective bactericides, so as to provide a theoretical basis for the control of soft rot of P. ternata. In this study, the pathogen was identified based on molecular biology and physiological biochemistry, followed by the detection of pathogenicity and pathogenicity spectrum via plant tissue inoculation in vitro and the indoor toxicity determination using the inhibition zone method to screen out bactericide with good antibacterial effects. The control effect of the bactericide against P. ternata soft rot was verified by the leave and tuber inoculation in vitro. The phylogenetic tree was constructed based on the 16 S rDNA, dnaX gene, and recA gene sequences, respectively, and the result showed that the pathogen belonged to the same branch as the type strain Dickeya fangzhongdai JS5. The physiological and biochemical tests showed that the pathogen was identical to D. fangzhongdai, which proved that the pathogen was D. fangzhongdai. The pathogenicity test indicated that the pathogen could obviously infect leaves at 24 h and tubers in 3 d. As revealed by the indoor toxicity test, 0.3% tetramycin, 5% allicin, and 80% ethylicin had good antibacterial activities, with EC_(50) values all less than 50 mg·L~(-1). Tests in tissues in vitro showed that 5% allicin exhibited the best control effect, followed by 0.3% tetramycin and 10% zhongshengmycin oligosaccharide, and their preventive effects were better than curative effects. Therefore, 5% allicin can be used as the preferred agent for the control of P. ternata soft rot, and 0.3% tetramycin and 10% zhongshengmycin oligosaccharide as the alternatives. This study has provided a certain theoretical basis for the control of P. ternata soft rot.
Phylogeny ; Pinellia/chemistry* ; Plant Leaves ; Plant Tubers

OBJECTIVE: To express and purify the antigenic peptide of adeno-associated virus (AAV) capsid conserved regions in prokaryotic cells and prepare its rabbit polyclonal antibody. METHODS: The DNA sequence encoding the conserved regions of AAV capsid protein was synthesized and cloned into the vector pET30a to obtain the plasmid pET30a-AAV-CR for prokaryotic expression and purification of the conserved peptides. Coomassie blue staining and Western blotting were used to identify the AAV conserved peptides. Japanese big ear white rabbits were immunized with AAV conserved region protein to prepare polyclonal antibody, with the rabbits injected with PBS as the control group. The antibody titer was determined with ELISA, and the performance of the antibody for recognizing capsid protein sequences of AAV1-AAV10 was assessed with Western blotting and immunofluorescence assay. RESULTS: The plasmid pET30a-AAV-CR was successfully constructed, and a recombinant protein with a relative molecular mass of 17000 was obtained. The purified protein induced the production of antibodies against the conserved regions of AAV capsid in rabbits, and the titer of the purified antibodies reached 1:320 000. The antibodies were capable of recognizing a wide range of capsid protein sequences of AAV1-AAV10. CONCLUSION We successfully obtained the polyclonal antibodies against AAV capsid conserved region protein from rabbits, which facilitate future studies of AAV vector development and the biological functions of AAV.
Animals ; Antibodies ; Capsid ; Capsid Proteins/genetics* ; Dependovirus/genetics* ; Prokaryotic Cells ; Rabbits ; Recombinant Proteins/genetics*