Anatomical variations, treatment and outcomes of Herlyn-Werner-Wunderlich syndrome: a literature review of 1673 cases.

PURPOSE: This study aimed to perform a systematic review of patients with Herlyn-Werner-Wunderlich syndrome (HWWS) and present the prevalence of symptoms, anatomical variants, endometriosis, surgical interventions, and short- and long-term outcomes. METHODS: A structured search was performed in PubMed, Scopus, Embase, and China National Knowledge Infrastructure, and studies published between 1 January, 2000 and 19 April, 2022 were included. The following data on HWWS were recorded: symptoms, anatomical variations, surgical interventions and short- and long-term outcomes. RESULTS: A total of 1673 patients were included in our analysis. The main symptoms were dysmenorrhea (53.8%), abnormal uterine bleeding (28.9%), and vaginal discharge (26.6%). The principal anomalies were right-obstructed hemivagina (57.3%), haematocolpos (81.7%), uterus didelphys (88.8%), and ipsilateral renal agenesis (93.1%). A majority of patients belonged to classification 1.1 (46.7%), with a blind hemivagina, and classification 2.1 (39.2%), with a small communication between two vaginas. The mainstay of treatment was vaginal septum excision (91.8%). Minimally invasive surgery (48.5%) was performed only after vaginal surgery (61.9%), and only a few patients required a second surgery (2.2%). Endometriosis was found in 9.6% of the patients. Fifty-two percent of them had ipsilateral ovarian endometriosis cysts. Pregnancy rate of these patients was 72.1%. The rate of adverse pregnancy outcomes was 22.4%. The caesarean section rate was 61.2%. CONCLUSIONS: Patients with HWWS presented with nonspecific symptoms and demonstrated various combinations of anomalies. The most common anatomical variants are classifications 1.1 and 2.1. Vaginal septum excision is effective in relieving symptoms and preventing complications, with hysteroscopic surgery as an option where there is concern about protecting the hymen from minor injury. The pregnancy rate for these patients after surgery was satisfactory, and the rate of adverse pregnancy outcomes after surgery was acceptable. We advise females with urological anomalies to be screened for Müllerian anomalies because of the close association between these two types of anomalies. Thus, HWWS contributes to the occurrence of endometriosis; however, more research is required to investigate the relationship between pelvic endometriosis and HWWS.

Sphingosine-1-phosphate receptor modulators in stroke treatment.

Sphingosine-1-phosphate (S1P) is a bioactive lysophospholipid that can influence a broad range of biological processes through its binding to five distinct G-protein-coupled receptors. S1P receptor modulators are a new group of immunosuppressive agents currently used in the immunotherapy of multiple sclerosis. Inflammation following stroke can exacerbate neuronal injury. Given that S1P signaling is linked to multiple immune processes, therapies targeting the S1P axis may be suitable for treating stroke. In this review, we outline S1P metabolism and S1P receptors, discuss the mechanisms of action of S1P receptor modulators in lymphocyte migration and their direct action on cells of the central nervous system, and provide a concise summary of the efficacy of S1P receptor modulators in animal studies and clinical trials on treatments for stroke.

Characterization and mutagenesis of a novel Mycobacterium smegmatis-derived glutamate decarboxylase active at neutral pH.

γ-aminobutyric acid (GABA) has various physiological functions and is widely used in medicine, food, and other fields. Glutamate decarboxylase (GAD) is a key enzyme that catalyzes the decarboxylation of L-glutamate to synthesize GABA. However, the industrial application of microorganism-derived GAD is limited by its rapid loss of enzymatic activity with pH approaching neutrality. In this study, a novel glutamate decarboxylase, GADMSM, from Mycobacterium smegmatis was overexpressed and purified. On the basis of homologous modeling and substrate molecular docking, several GADMSM mutants were constructed, and their enzymatic properties were analyzed. The results showed that the optimal pH of wild-type GADMSM is 5.4; at pH 6.2, 22.8% enzymatic activity was retained. The T211I replacement in GAD and C-terminal deletion mutant GADMSMΔC showed relatively high catalytic activity in a pH range of 5.0-7.0. The Vmax and Km values of GADMSMΔC were 14.69 and 5.70, respectively, at pH 5.5, and 9.87 and 6.17, respectively, at pH 7.0. Compared with the wild-type GAD, GADMSMΔC maintained higher affinity and enzymatic activity of the substrate, maintaining 78.5% of the highest enzymatic activity even at pH 7.0, which is the highest reported activity retention for GAD under neutral pH condition. Therefore, GADMSMΔC can be used for the transformation of high-yielding strains and industrial production of GABA.

circ0000069 promotes cervical cancer cell proliferation and migration by inhibiting miR-4426.

Circular RNAs (circRNAs) have been shown to be associated with the occurrence and development of cervical cancer (CC). In the present study, we aimed to investigate the tumor-promoting effect of hsa_circ_0000069 (circ0000069) on CC and the mechanisms underlying its effect. We found that circ0000069 was upregulated in CC cells and tissues, and that N6-methyladenosine (m6A) modification maintained circ0000069 stability. Gain- and loss-of-function assays revealed that circ0000069 promoted CC cell proliferation and migration. miR-4426 specifically binds circ0000069 and mediates its functions in CC development. In conclusion, circ0000069 was upregulated partially due to m6A modification, which promoted cell proliferation and migration via sponging miR-4426 in CC.

Rapid and label free detection of aflatoxin B1 in alcoholic beverages with a microfluid fiber device.

A rapid and label free aflatoxin B1 (AFB1) microfluid sensor was proposed and tested. The device was fabricated with hollow-core photonics crystal fiber infiltrated with the AFB1 solution. The autofluorescence emitting from the AFB1 molecules was detected. The sensor length was optimized. The AFB1 concentration was tested with a 4 cm long sensor. The best limit of detection was achieved as low as 1.34 ng/ml, which meets the test requirement of the national standards for AFB1 in food. The effectiveness of this sensor being applied in beer solution was also verified to be a little more sensitive than in aqueous solution. Compared with traditional AFB1 detection methods, the proposed single-ended device perfectly satisfies the demand of process control in alcoholic beverages manufacture.

vNOTES Hysterectomy with Sentinel Lymph Node Mapping for Endometrial Cancer: Description of Technique and Perioperative Outcomes.

STUDY OBJECTIVE: To explore the technique and clinical value of transvaginal natural orifice transluminal endoscopic surgery (vNOTES) in hysterectomy and sentinel lymph node (SLN) mapping for endometrial cancer by comparing its perioperative outcomes with those of laparoscopic staging. DESIGN: Retrospective cohort study. SETTING: Department of gynecology at a tertiary medical center. PATIENTS: All women diagnosed with endometrial cancer who underwent minimally invasive surgery at our center between August 2017 and May 2020. INTERVENTIONS: Both vNOTES and laparoscopic approaches were used for hysterectomy and SLN mapping. The success of SLN detection as well as perioperative outcomes were subsequently analyzed. MEASUREMENTS AND MAIN RESULTS: This study included 74 patients; 23 patients underwent vNOTES surgery, whereas 51 underwent standard laparoscopic surgery. The total successful SLN detection was 95.7% in the vNOTES group and 92.2% in the laparoscopy group (p >.05), whereas the bilateral success rates were 87.0% and 90.2%, respectively. No difference in SLN detection was observed between the 2 groups in terms of the side-specific mapping efficacy quotient (91.3% vs 91.2%, p = .47). The number of harvested SLNs, operative time, estimated blood loss, and intraoperative complications in the 2 groups were similar. One (4.3%) postoperative complication occurred in the vNOTES group vs 4 (7.9%) in the laparoscopy group (p = .029), and the median postoperative hospital stay was 3 days vs 4 days (p = .003). CONCLUSION: This study suggests that the vNOTES procedure is feasible, with a potentially decreased postoperative hospital stay, faster recovery, and better cosmetic results. However, prospective research is needed to validate its broader clinical application.

Automated and accurate assessment for postural abnormalities in patients with Parkinson's disease based on Kinect and machine learning.

BACKGROUND: Automated and accurate assessment for postural abnormalities is necessary to monitor the clinical progress of Parkinson's disease (PD). The combination of depth camera and machine learning makes this purpose possible. METHODS: Kinect was used to collect the postural images from 70 PD patients. The collected images were processed to extract three-dimensional body joints, which were then converted to two-dimensional body joints to obtain eight quantified coronal and sagittal features (F1-F8) of the trunk. The decision tree classifier was carried out over a data set established by the collected features and the corresponding doctors' MDS-UPDRS-III 3.13 (the 13th item of the third part of Movement Disorder Society-Sponsored Revision of the Unified Parkinson's Disease Rating Scale) scores. An objective function was implanted to further improve the human-machine consistency. RESULTS: The automated grading of postural abnormalities for PD patients was realized with only six selected features. The intraclass correlation coefficient (ICC) between the machine's and doctors' score was 0.940 (95%CI, 0.905-0.962), meaning the machine was highly consistent with the doctors' judgement. Besides, the decision tree classifier performed outstandingly, reaching 90.0% of accuracy, 95.7% of specificity and 89.1% of sensitivity in rating postural severity. CONCLUSIONS: We developed an intelligent evaluation system to provide accurate and automated assessment of trunk postural abnormalities in PD patients. This study demonstrates the practicability of our proposed method in the clinical scenario to help making the medical decision about PD.

Protective Effects of Nucleosides-Rich Extract from Cordyceps cicadae against Cisplatin Induced Testicular Damage.

Cisplatin (CISP) is an efficacious anticancer agent used in chemotherapy, however, the constraint to its clinical utility is the stray organ toxicity including testicular damage linked to oxidative and inflammatory cascades. This study aimed to explore the protective effect of nucleosides-rich extract from Cordyceps cicadae (NRCE) against CISP-induced testicular damage in rats. Rats were subjected to prophylactic oral administration of NRCE (50, 100 and 400 mg/kg body weight/day) for 7 days prior to testicular toxicity induced by CISP (10 mg/kg, ip) and were sacrificed after 72 h post-CISP injection. Cisplatin caused significant deficits in sperm count, viability and motility, testosterone and follicle stimulating hormone (FSH) compared to normal control. It depressed testicular activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), total antioxidant content (TAC), whereas malondialdehyde (MDA) increased remarkably. CISP considerably increased tumor necrosis factor-alpha (TNF-α) and interleukin-one beta (IL-1ß) with alterations in testis histology compared to normal control. Interestingly, NRCE pretreatment inhibited the CISP-induced alterations in reproductive indices, restored the antioxidant activities in testes as well as inflammatory mediators and histology comparable to control. Our findings demonstrate that NRCE could prevent CISP testicular damage via inhibition of oxidative stress and pro-inflammation in rats.

Tourniquet device for hemorrhage control during cesarean section of complete placenta previa pregnancies.

AIM: To reduce massive and potentially fatal hemorrhage during cesarean section (CS) delivery because of the transverse incision through the lower uterine segment and placenta in central placenta previa (CPP), especially the type in which the placenta attaches to the anterior wall of the uterus. METHODS: Thirty-four patients with CPP were enrolled in the study and 14 of them accepted the tourniquet method. The main outcome evaluations were blood loss, the proportion of red blood cell-transfused patients, the need for other hemostatic procedures and complications in either the mother or infant after the completion of the tourniquet procedure. RESULTS: A statistically significant benefit was noted between the two groups of the frequency of post-partum hemorrhage (PPH), the median estimated blood loss and the proportion of transfused patients. The effectiveness of the tourniquet method could be further proved by the relative risk with confidence intervals for PPH data and additional procedures. CONCLUSION: The clinical data of 34 patients show that this is an effective surgical treatment to reduce blood loss and suggest that there is negligible harm to patients and fetuses.

The anti-colorectal cancer effect and metabolites of Agrimonia pilosa Ledeb.

ETHNOPHARMACOLOGICAL RELEVANCE: Agrimonia pilosa Ledeb. (Rosaceae, A. pilosa) has been used in traditional medicine in China, Japan, Korea, and other Asian countries for treatment of acute and chronic enteritis and diarrhea. Secondary metabolites have been isolated and tested for biological activities. It remains unclear in terms of its potential components of anti-colorectal cancer properties. AIM OF THE STUDY: The study aimed to how extracts from A. pilosa and their components influenced tumor microenvironment and the colorectal tumor growth in vivo on AOM/DSS induced colorectal cancer mice, the metabolites of A. pilosa was also been studied. MATERIALS AND METHODS: Different methods have been used to extract different parts of A. pilosa. And the anti-proliferation effect of these extracts on colon cancer cells have been tested. The components of A. pilosa and its metabolites in vivo were analyzed by UPLC-QTOF-MS/MS. The anti-colorectal cancer (CRC) effects of A. pilosa and its components in vivo were studied on AOM/DSS induced CRC mice. The effects of constituents of A. pilosa on the composition of immune cells in tumor microenvironment (TME) were analyzed by flow cytometry. 16 S rDNA technology was used to analyze the effect of administration on the composition of intestinal microflora. Pathological section staining was used to compare the morphological changes and molecular expression of intestinal tissue in different groups. RESULTS: The constituent exists in root of A. pilosa showed the strongest anti-proliferation ability on colon cancer cells in vitro. The extract from the root of A. pilosa could attenuate the occurrence of colorectal tumors induced by AOM/DSS in a concentration-dependent manner. Administration of the extract from the root of A. pilosa could affect the proportion of γδT cells, tumor associated macrophages and myeloid derived suppressor cells in TME, increasing the proportion of anti-tumor immune cells and decrease the immunosuppressive cells in the TME to promote the anti-tumor immune response. The administration of the extract adjusted the composition of gut microbiota and its components Agrimoniin and Agrimonolide-6-o-glucoside showed the strongest anti-CRC effect in vivo with adjusting the gut microbiota differently. CONCLUSIONS: The extract from root of A. pilosa showed anti-colorectal cancer effects in vivo and in vitro, affecting the composition of gut microbiota and the anti-tumor immune response. Within all components of A. pilosa, Agrimoniin and Agrimonolide-6-o-glucoside showed remarkable anti-CRC efficiency in vivo and in vitro. Besides, the metabolites of extract from root of A. pilosa in gastrointestinal tract mainly composed of two parts: Agrimonolide-related metabolites and Urolithins. The extract from root of A. pilosa could contribute to potential drugs for assisting clinical anti-colon cancer therapy.

Prospective randomized comparison of laparoscopic peritoneal vaginoplasty with laparoscopic sigmoid vaginoplasty for treating congenital vaginal agenesis.

INTRODUCTION AND HYPOTHESIS: The aim of this study was to compare the effectiveness and long-term anatomic and functional results of laparoscopic peritoneal vaginoplasty and laparoscopic sigmoid vaginoplasty. METHODS: From January 2002 to December 2010, 40 patients with congenital vaginal agenesis were prospectively randomized to undergo either laparoscopic peritoneal vaginoplasty (26 cases) or laparoscopic sigmoid vaginoplasty (14 cases) in 2:1 ratio. Pre- and postoperative examination findings, Female Sexual Function Index (FSFI) questionnaire responses, and sexual satisfaction rates are reported. RESULTS: All surgical procedures were performed successfully, with no intraoperative complications. The laparoscopic peritoneal vaginoplasty group had significantly less blood loss and a surgery shorter on average than the laparoscopic sigmoid colovaginoplasty group. Postoperative course was uneventful for all patients in both groups, though postoperative retention time and hospital stay were less for peritoneal vaginoplasty patients than for sigmoid vaginoplasty patients. Mean neovaginal length, excessive mucous production, sexual life initiation time, and sexual satisfaction rate were similar between groups. Patient complaints of abdominal discomfort, unusual odor from vaginal secretions, and vaginal contraction during intercourse were higher in the sigmoid colovaginoplasty group (p < 0.005 vs. peritoneal vaginoplasty). Postoperative FSFI scores did not differ significantly between groups. CONCLUSIONS: Relative to laparoscopic sigmoid colovaginoplasty, laparoscopic peritoneal vaginoplasty provides good anatomic and functional results and excellent patient satisfaction.

The efficacy of ureteroscopic triage in increasing the cure rate of the first-line treatment for a ureterovaginal fistula resulting from radical hysterectomy.

Study objective: To explore the effect of pretreatment with ureteroscopic triage for iatrogenic ureterovaginal fistula (UVF) resulting from radical hysterectomy. Design: A retrospective cohort study. Setting: Department of gynecology at a tertiary medical center. Patients: Women diagnosed with UVF secondary to radical hysterectomy at our center between April 2008 to June 2018. Interventions: The patients were divided into two groups according to whether pretreatment with ureteroscopic triage was performed. Those in the non-triage group underwent retrograde placement of a double-J stent immediately following diagnosis as the first-line therapy. Patients in the triage group were first evaluated under ureteroscopy, their ureteral injuries were then classified into different grades and then underwent different treatments as the first-line therapy, including stent placement or reconstruction surgeries. The cure rate of the first-line therapy and the timeliness of the implementation of adjuvant radiotherapy were subsequently analyzed. Measurements and main results: Ninety-eight UVF patients were included. The demographics, ECOG status, stage of cervical cancer (FIGO 2009), types and onset time of symptoms were not different between the two groups. There were 54 patients in the nontriage group, with an overall first-line cure rate of 70.4% and a timely implementation rate of adjuvant radiotherapy of 38.5%. There were 44 patients in the evaluation group, with an overall first-line cure rate of 93.2% and a timely implementation rate of adjuvant radiotherapy of 90.0%. The differences were statistically significant (p < 0.001). Conclusion: Ureteroscopic triage of ureteral injuries can guide the selection of the optimal first-line therapy for patients with UVF secondary to radical hysterectomy, increase the cure rate and ensure the timely implementation of adjuvant radiotherapy.

Coronavirus disease 2019 and acute cerebrovascular events: a comprehensive overview.

Since the Corona Virus Disease 2019 (COVID-19) pandemic, there has been increasing evidence that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is associated with acute cerebrovascular events such as cerebral infarction, cerebral hemorrhage, and cerebral venous thrombosis. Although the mechanism of cerebrovascular complications among COVID-19 patients has not been adequately elucidated, the hypercoagulable state, excessive inflammation and ACE-2-associated alterations in the renin-angiotensin-aldosterone system after SARS-CoV-2 infection probably play an essential role. In this overview, we discuss the possible mechanisms underlying the SARS-CoV-2 infection leading to acute cerebrovascular events and review the characteristics of COVID-19-related acute cerebrovascular events cases and treatment options available worldwide.

Extinction risk modeling predicts range-wide differences of climate change impact on Karner blue butterfly (Lycaeides melissa samuelis).

The Karner blue butterfly (Lycaeides melissa samuelis, or Kbb), a federally endangered species under the U.S. Endangered Species Act in decline due to habitat loss, can be further threatened by climate change. Evaluating how climate shapes the population trend of the Kbb can help in the development of adaptive management plans. Current demographic models for the Kbb incorporate in either a density-dependent or density-independent manner. We instead created mixed density-dependent and -independent (hereafter "endo-exogenous") models for Kbbs based on long-term count data of five isolated populations in the upper Midwest, United States during two flight periods (May to June and July to August) to understand how the growth rates were related to previous population densities and abiotic environmental conditions, including various macro- and micro-climatic variables. Our endo-exogenous extinction risk models showed that both density-dependent and -independent components were vital drivers of the historical population trends. However, climate change impacts were not always detrimental to Kbbs. Despite the decrease of population growth rate with higher overwinter temperatures and spring precipitations in the first generation, the growth rate increased with higher summer temperatures and precipitations in the second generation. We concluded that finer spatiotemporally scaled models could be more rewarding in guiding the decision-making process of Kbb restoration under climate change.

Deficiency of SR-B1 reduced the tumor load of colitis-induced or APCmin /+ -induced colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is one of the most common tumors in the world. Cholesterol plays an important role in the pathogenesis of tumors. One of the cholesterol transporters, scavenger receptor class B type 1 (SR-B1), a multi-ligand membrane receptor protein, is expressed in the intestines which also highly expressed in various tumors. But the potential mechanism of SR-B1 in CRC development has not been reported. AIMS: This study aimed to clarify the importance of SR-B1 in the development and prognosis of CRC as much as possible to provide a possible strategy in CRC treatment. MATERIALS & METHODS: In this study, we used SR-B1 gene knockdown mice to study the effect of SR-B1 on colitis-induced or APCmin/+ -induced CRC. The expression of related molecules were detected through the immunohistochemistry and hematoxylin-eosin staining, western blot analysis, and Flow cytometry. The gene expression and microbiota in microenvironment of CRC mice were analyzed through eukaryotic mRNA sequencing and 16S rRNA high-throughput sequencing. RESULTS: The results showed that SR-B1 knockdown reduced the tumor load of colitis-induced or APCmin/+ -induced CRC. SR-B1 knockdown improved the immune microenvironment by affecting the level of tumor-associated macrophage (TAM), mononuclear myeloid-derived suppressor cells (M-MDSCs), granulocytic myeloid-derived suppressor cells (G-MDSCs), programmed cell death-ligand 1 (PD-L1), and human leukocyte antigen class I-B (HLA-B), and also reduced the level of low-density lipoprotein receptor (LDL-R), and increased the level of ATP binding cassette transporter A1 (ABCA1) to regulate the cholesterol metabolism, and regulated the expression of related genes and intestinal microbiota. SR-B1 knockdown can also trigger the anti-CRC effect of anti-PD 1 in colitis-induced CRC. DISCUSSION: SR-B1 deficiency significantly improved the immunity in tumor microenvironment of colitis-induced or APCmin/+ -induced CRC. In addition, the microbiota changes caused by SR-B1 deficiency favor improving the immune response to chemotherapeutic drugs and anti-PD1 therapy. The mechanism of action of SR-B1 deficiency on the development of CRC still needs further in-depth research. CONCLUSION: This study provides a new treatment strategy for treating CRC by affecting the expression of SR-B1 in intestine.

Altered expression of circular RNA in patients with cervical artery dissection.

Cervical artery dissection (CeAD), a special cerebrovascular disease and the main cause of stroke in young people, can present with ischemic stroke, headache, subarachnoid hemorrhage, and other symptoms, increasing the possibility of misdiagnosis. As a special class of non-coding RNAs, circRNAs are commonly found in organisms and can play regulatory roles in transcription and post-transcription processes, affecting gene expression.CircRNAs have reported to be associated with neurological diseases; however, their role in CeAD has not been discerned. In this study, we aimed to elucidate the pathophysiological changes in patients with CeAD and identify biomarkers. Peripheral blood mononuclear cells from patients with CeAD and healthy controls were sequenced using high-throughput sequencing. We detected 460 differently expressed circRNAs in patients with CeAD (p < 0.5, fold difference ≥ 2), of which 240 were upregulated and 220 were downregulated. Four circRNAs showed significant differences in expression, which were validated using qRT-PCR. These results suggested that three circRNAs were consistent with high-throughput sequencing results. Bioinformatics analysis demonstrated that these differentially expressed circRNAs were involved in protein metabolism, regulation, synapses, and other pathophysiological processes during CeAD-induced stroke. Additionally, various pathways related to inflammation were closely associated with circRNAs. Based on our results, we suggest that the aberrant expression of circRNAs in CeAD may serve as a biomarker for its diagnosis and as a potential therapeutic target.

Phosphoproteomic Analysis Reveals the Predominating Cellular Processes and the Involved Key Phosphoproteins Essential for the Proliferation of Toxoplasma gondii.

PURPOSE: To explore the essential roles of phosphorylation in mediating the proliferation of T. gondii in its cell lytic life. METHODS: We profiled the phosphoproteome data of T. gondii residing in HFF cells for 2 h and 6 h, representing the early- and late-stages of proliferation (ESP and LSP) within its first generation of division. RESULTS: We identified 70 phosphoproteins, among which 8 phosphoproteins were quantified with the phosphorylation level significantly regulated. While only two of the eight phosphoproteins, GRA7 and TGGT1_242070, were significantly down-regulated at the transcriptional level in the group of LSP vs. ESP. Moreover, GO terms correlated with host membrane component were significantly enriched in the category of cellular component, suggesting phosphoprotein played important roles in acquiring essential substance from host cell via manipulating host membrane. Further GO analysis in the categories of molecular function and biological process and pathway analysis revealed that the cellular processes of glucose and lipid metabolism were regulated by T. gondii phosphoproteins such as PMCAA1, LIPIN, Pyk1 and ALD. Additionally, several phosphoproteins were enriched at the central nodes in the protein-protein interaction network, which may have essential roles in T. gondii proliferation including GAP45, MLC1, fructose-1,6-bisphosphate aldolase, GRAs and so on. CONCLUSION: This study revealed the main cellular processes and key phosphoproteins crucial for the intracellular proliferation of T. gondii, which would provide clues to explore the roles of phosphorylation in regulating the development of tachyzoites and new insight into the mechanism of T. gondii development in vitro.

Genome editing mRNA nanotherapies inhibit cervical cancer progression and regulate the immunosuppressive microenvironment for adoptive T-cell therapy.

CRISPR/Cas9-based genome editing is promising for therapy of cervical cancer by precisely targeting human papillomavirus (HPV). To develop CRISPR/Cas9-based genome editing nanotherapies, a pH-responsive hybrid nonviral nanovector was constructed for co-delivering Cas9 mRNA and guide RNAs (gRNAs) targeting E6 or E7 oncogenes. The pH-responsive nanovector was fabricated using an acetalated cyclic oligosaccharide (ACD), in combination with low molecular weight polyethyleneimine. Thus obtained hybrid ACD nanoparticles (defined as ACD NP) showed efficient loading for both Cas9 mRNA and E6 or E7 gRNA, giving rise to two pH-responsive genome editing nanotherapies E6/ACD NP and E7/ACD NP, respectively. Cellularly, ACD NP exhibited high transfection but low cytotoxicity in HeLa cervical carcinoma cells. Also, efficient genome editing of target genes was achieved in HeLa cells, with minimal off-target effects. In mice bearing HeLa xenografts, treatment with E6/ACD NP or E7/ACD NP afforded effective editing of target oncogenes and considerable antitumor activities. More importantly, treatment with E6/ACD NP or E7/ACD NP notably promoted CD8+ T cell survival by reversing the immunosuppressive microenvironment, thereby leading to synergistic antitumor effects by combination therapy using the gene editing nanotherapies and adoptive T-cell transfer. Consequently, our pH-responsive genome editing nanotherapies deserve further development for the treatment of HPV-associated cervical cancer, and they can also serve as promising nanotherapies to improve efficacies of other immune therapies against different advanced cancers by regulating the immunosuppressive tumor microenvironment.

Learning curve for laparoendoscopic single-site radical hysterectomy using the "chopstick" technique: a retrospective cohort study.

Background: Laparoendoscopic single-site surgery (LESS), as a promising minimally invasive surgery, is confronted with the obstacle of the "chopstick effect" which limits its further application. The "chopstick" technique is characterized by the usage of instruments of parallel and equal length, and in operations relying on double-fulcrum and unique surgeon position can play a key role in overcoming the above disadvantage effect. This study sought to explore the learning curve for the use of the novel "chopstick" technique in laparoendoscopic single-site radical hysterectomy (LESS-RH) and evaluate the technique's practicability. Methods: Consecutive cervical cancer patients who underwent LESS-RH with the "chopstick" technique by a surgeon with rich experience in laparoscopy from November 2016 to September 2018 were included in the study. The learning curve of his surgeries with the "chopstick" technique was evaluated using the cumulative summation (CUSUM) method by analyzing operation time (OT) which was the surrogate indicator of surgical ability. The patients were divided into Phase I and Phase II based on the learning curve peak, whose demographic and perioperative characteristics, such as tumor Federation International of Gynecology and Obstetrics (FIGO) stage and histology, operation time, blood loss, and complications were collected and analyzed. Results: The mean OT was 231.5 min (range, 115-355 min). The division of the learning curve based on OT occurred after the first 15 cases were finished, dividing Phase I and Phase II. The mean OT for Phase I (259 min) was significantly longer than that of Phase II (219 min) (P=0.02). Only 1 intraoperative complication occurred in Phase I, and none occurred in Phase II. Major postoperative complications occurred more frequently in Phase I (N=3) than in Phase II (N=0). No significant differences were observed in terms of lymph nodes, blood loss, or pathological features. Conclusions: The "chopstick" technique may help surgeons obtain stable LESS surgical performance through a relatively short learning curve, even in some complex surgeries, such as radical hysterectomy.

The Alteration Profiles of m6A-Tagged circRNAs in the Peri-Infarct Cortex After Cerebral Ischemia in Mice.

The N6-methyladenosine (m6A) modification acts as a dynamic regulatory factor in diseases by regulating the metabolism and function of the transcriptome, especially mRNAs. However, little is known regarding the functional effects of m6A modifications on circRNAs. In this research, we established a distal middle cerebral artery occlusion (MCAO) model in adult C57BL/6J mice. The mice were divided into three groups: sham surgery, 3 days after MCAO (3d), and 7 days after MCAO (7d). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) demonstrated that the mRNA expression levels of m6A-related methyltransferases (METTL3, METTL14), demethylases (FTO, ALKBH5), and reading proteins (YTHDF1, YTHDF3) altered compared to the sham group. Furthermore, the translation level of ALKBH5 and YTHDF3 was significantly decreased in the 3d group while increased in 7d group. Methylated RNA immunoprecipitation (MeRIP) and circRNA microarray indicated 85 hypermethylated and 1621 hypomethylated circRNAs in the 3d group. In the 7d group, the methylation level increased in 57 and decreased in 66 circRNAs. Subsequently, our results were verified by MeRIP-qPCR. Bioinformatics analysis was performed to analyze the functions of differentially m6A-modified circRNAs. We found some m6A modified-circRNAs associated with cerebral infarction, providing a new direction for the molecular mechanism of stroke.