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Purpose To identify the clinical features of intra-articular osteoid osteoma (OO) of the hip, to evaluate the clinical effect of arthroscopic excision for intra-articular OO, and to summarize the characteristics of revision cases of hip OO and the revision surgery under arthroscopy in these cases. Methods We retrospectively reviewed the data of 25 patients who underwent arthroscopic excision of hip OO. The case series included 10 patients who underwent revision surgery. Lesion location, presenting symptoms, and symptom duration were analyzed; postoperative improvement was assessed using the modified Harris Hip Score (mHHS) and International Hip Outcomes Tool (iHot-12) score. We examined the reasons for revision surgery and the characteristics of OO progression after the first surgery. Results The most common presenting symptom was groin pain that was relieved by nonsteroidal anti-inflammatory drugs (NSAIDs). Varying degrees of limitation of range of motion (ROM) were present in all patients. The osteosclerosis around the tumor nest on computed tomography (CT) scan is a characteristic radiographic feature in this disease. However, the classic radiographic feature was apparent on plain x-rays in only 2 of 25 patients. As a kind of efficient radiological method, magnetic resonance imaging (MRI) can help in distinguishing OO from femoroacetabular impingement (FAI), as the latter is characterized by a large effusion and bone marrow edema at the atypical site of impingement. For the patients who had only 1 arthroscopic resection, the mean (± standard deviation) mHHS and iHot-12 scores were 70.30 ± 9.06 (range 51 to 86) and 75.07 ± 7.69 (57 to 88), respectively. At last follow-up, the mean scores were 98.30 ± 2.15 (94 to 100) and 97.76 ± 2.04 (94 to 100). For revision cases, the mean mHHS and iHot-12 scores were 68.55 ± 3.77 (60 to 72) and 67.88 ± 5.39 (56 to 76). At last follow-up, the mean scores were 97.11 ± 2.47 (94 to 100) and 95.22 ± 1.78 (94 to 100). In the present study, 24 of 25 patients (96%) reached the minimal clinically important difference (MCID) of mHHS, and 21 of 22 patients (95.2%) reached the MCID of iHot-12. Among the revision patients, the most common misdiagnosis at first surgery was FAI. Another feature is that a wrong diagnosis or incomplete intra-articular OO resection can stimulate the tumor and cause an inflammatory reaction and rapidly progressive OA, necessitating prompt revision surgery for complete removal. The degree of joint degeneration was related to the time since the first operation. Conclusion OO of the hip joint typically presents with pain and limited joint activity. Misdiagnosis as FAI or synovitis is common, and CT scan is very helpful for accuracy diagnosis. Arthroscopic excision appears to be an effective method for the treatment of OO of the hip joint. Level of Evidence IV, case series.

α-Synuclein (α-syn), especially its abnormal oligomeric and phosphorylated form, plays a critical role in the pathogenesis of Parkinson’s disease (PD). Plasma exosomal α-syn species have been shown to be a promising PD biomarker. However, whether different α-syn species in plasma exosomes (the oligomeric α-syn and the Ser129 phosphorylated α-syn (p-α-syn)) which represent the PD pathogenesis in the brain could be specific peripheral PD biomarker haven’t been well elucidated. In this study, we successfully extracted and identified the human plasma exosomes, and the CNS-derived exosomes were detected. The different aggregation status, localization and degradation characteristics of α-syn and p-α-syn in the plasma exosomes between PD patients and healthy controls were further analyzed. The results suggested that α-syn and p-α-syn in the plasma exosomes of PD patients showed poor solubility after protease K (PK) treatment. Aggregated α-syn and p-α-syn existed both inside and on the membrane surface of plasma exosomes. The Receiver operating characteristic (ROC) performance of α-syn oligomer/total α-syn in exosomes was moderately helpful in PD diagnosis (AUC = 0.71, sensitivity = 60.5%, specificity = 59.4%), and the ratio of p-α-syn oligomer/total p-α-syn showed similar result (AUC = 0.69, sensitivity = 60.0%, specificity = 59.5%). This study indicates that the oligomeric α-syn/total α-syn and oligomeric p-α-syn/total p-α-syn ratio in plasma exosomes may be applied to assist the PD diagnosis, which needs further research.

To compare the oncological outcomes and major complications of laparoscopic radical hysterectomy (LRH) and abdominal radical hysterectomy (ARH) for stage IB1 cervical cancer (FIGO 2009) with a tumour size less than 2 cm.We retrospectively compared the oncological outcomes and major complications of 1207 stage IB1 cervical cancer patients with a tumour size less than 2 cm who received LRH (n = 546) or ARH (n = 661) in 37 hospitals.(1) There was no significant difference in 3-year overall survival (OS; 97.3% vs. 98.5%, P = 0.288) or 3-year disease-free survival (DFS; 95.1% vs. 95.4%, P = 0.792) between LRH (n = 546) and ARH (n = 661).(2) The rate of any 1 complication refers to the incidence of one or more complications in a patient, which was higher with LRH than ARH (OR = 4.047, 95% CI = 2.035-8.048, P 0.001). Additionally, intraoperative complications occurred with LRH (OR = 12.313, 95% confidence intervals [CI] = 1.571-96.493, P = 0.017), and postoperative complications (OR = 3.652, 95% CI = 1.763-7.562, P 0.001) were higher with LRH than ARH. The ureteral injury rate was higher with LRH than with ARH (1.50% vs. 0.20%, OR = 9.814, 95% CI = 1.224-78.712, P = 0.032). The ureterovaginal fistula rate was higher with LRH than ARH. The rates of obturator nerve injury, bladder injury, vesicovaginal fistula, rectovaginal fistula, venous thromboembolism, bowel obstruction, chylous leakage, pelvic haematoma, and haemorrhage were similar between the groups.The oncological outcomes of LRH and ARH for stage IB1 cervical cancer patients with a tumour size less than 2 cm do not differ significantly. However, incidences of any 1 complication, intraoperative complications, and postoperative complications were higher with LRH than ARH, with complications manifesting mainly as ureteral injury and uterovaginal fistula.

BACKGROUND: China’s atmospheric PM2.5 pollution is serious, and PM2.5 exerts a negative impact on the human respiratory system, cardiovascular, and mental health, and even more serious health risk for the elderly with weak immunity. OBJECTIVE: This work aims to analyse the impacts of PM2.5 microenvironment exposure on the health of the elderly and provide corresponding countermeasures. METHODS: The survey subjects are 118 retired elderly people in the community. PM2.5 exposure concentrations are monitored in summer (June 10 ∼ July 10, 2019) and winter (November 25 ∼ December 25, 2019). RESULTS: The exposure concentration in winter is higher than that in summer, with statistical difference (P < 0.05). Under the impact of PM2.5 microenvironment exposure, smoking in the elderly can increase the concentration of PM2.5, and long-term exposure to PM2.5 in the elderly can cause mental health problems. CONCLUSION: Long-term exposure of the elderly to the PM2.5 microenvironment leads to physical diseases and even psychological problems, which requires attention.

The purpose of this study was to investigate the surgical methods and clinical outcomes of arthroscopic treatment of a special type of calcification with surrounding inflammation in the acetabular labrum of the hip, which was temporarily named “calcifying labrumitis”. From April 2015 to November 2019, a total of seven patients with calcifying labrumitis of the hip who underwent arthroscopic excision of calcified lesions and suture or partial resection of the labrum were included in this study. Radiographs were retrospectively evaluated for morphologic characteristics of calcifying labrumitis. Each patient was assessed by the visual analogue scale (VAS), modified Harris hip score (mHHS), nonarthritic hip score (NAHS) and satisfaction rate before surgery and at the final follow-up evaluation. Seven patients, one male and six females aged 29–48 years, were included in the study; of these patients, three had calcifying labrumitis on the left side and four had calcifying labrumitis on the right side. All patients had hip pain and limited range of motion for a mean of 7.5 ± 3.1 months (range, 3–12 months). The mean follow-up period was 34.9 ± 19.5 months (range, 12–66 months). The lateral central-edge angle (LCEA) was 31.7 ± 3.9° (range, 28.8–36.4°), and the α angle was 41.4 ± 5.3° (range, 33.6–48.2°). None of the patients had cam or pincer lesions. After complete removal of calcified lesions, five patients underwent repair of the labrum with a suture anchor, and two patients underwent partial resection of the labrum. The symptoms of all patients improved significantly at the last follow-up. Mean scores improved from 5.8 ± 1.5 to 1.1 ± 0.3 (p

Objective to explore the differences in the central response after acupuncture and moxibusbution at Zusānlĭ (足三里ST36) in treatment of functional dyspepsia (FD) based on the analysis of fractional amplitude of low frequency fluctuation (fALFF) of resting-state functional magnetic resonance imaging (rfMRI). Methods a total of 60 patients with FD were randomized into an acupuncture group and a moxibustion group, 30 cases in each one. In the acupuncture group, acupuncture was applied to bilateral ST36. In the moxibustion group, moxibustion was exerted at bilateral ST36. The treatment was given once a day, 5 times a week, totally for 4 weeks in each group, including 20 treatments with acupuncture or moxibustion. Separately, before and at the end of treatment, rfMRI scanning was conducted in two groups. Using data processing assistant for resting-state fMRI (DPARSF) software and MATLAB data platform, rfMRI data were collected for preprocessing and fALFF analysis. Results compared with the data before treatment, after treated with acupuncture at ST36 in FD patients, fALFF value was reduced in the right superior frontal gyrus, left superior frontal gyrus, left inferior frontal gyrus, right cuneus, left precuneus, right middle occipital gyrus, left middle occipital gyrus, etc., and it was increased in the left parahippocampus, right parahippocampus, left cerebellum, etc (all P Conclusion the differences in central function responses are induced in treamtent with acupuncture and moxibustion, which is probably related to the resting-state default network associated with targeted regulation and ventral attention of brain areas in functional dyspepsia.

Background Thermotolerant yeast has outstanding potential in industrial applications. Komagataella phaffii (Pichia pastoris) is a common cell factory for industrial production of heterologous proteins. Results Herein, we obtained a thermotolerant K. phaffii mutant G14 by mutagenesis and adaptive evolution. G14 exhibited oxidative and thermal stress cross-tolerance and high heterologous protein production efficiency. The reactive oxygen species (ROS) level and lipid peroxidation in G14 were reduced compared to the parent. Oxidative stress response (OSR) and heat shock response (HSR) are two major responses to thermal stress, but the activation of them was different in G14 and its parent. Compared with the parent, G14 acquired the better performance owing to its stronger OSR. Peroxisomes, as the main cellular site for cellular ROS generation and detoxification, had larger volume in G14 than the parent. And, the peroxisomal catalase activity and expression level in G14 was also higher than that of the parent. Excitingly, the gene knockdown of CAT encoding peroxisomal catalase by dCas9 severely reduced the oxidative and thermal stress cross-tolerance of G14. These results suggested that the augmented OSR was responsible for the oxidative and thermal stress cross-tolerance of G14. Nevertheless, OSR was not strong enough to protect the parent from thermal stress, even when HSR was initiated. Therefore, the parent cannot recover, thereby inducing the autophagy pathway and resulting in severe cell death. Conclusions Our findings indicate the importance of peroxisome and the significance of redox balance in thermotolerance of yeasts.

ObjectiveTo explore the impact of the dietary patterns of truck drivers on their driving behaviours and the mediation effect of fatigue between these factors.MethodsA sample of 389 male truck drivers from a transport company in Suzhou, China completed the Food Frequency Questionnaire (FFQ), the Multidimensional Fatigue Inventory (MFI), the Positive Driver Behaviours Scale (PDBS) and the Driver Behaviour Questionnaire (DBQ). The associations among dietary patterns, fatigue and driving behaviour were examined using pathway analysis.ResultsFour dietary patterns were identified based on principal component analysis: animal-derived foods, staple foods, snacks and vegetables. The pathway analysis showed that the vegetable-rich pattern had a direct positive impact on positive driving behaviour (β=0.211, pβ=0.094, pβ=0.071, pβ=−0.191, pConclusionsOverall, the driving behaviours of truck drivers are correlated with their dietary patterns. Drivers who preferred vegetables and staple foods had more positive driving behaviour, while the animal-derived food and snack patterns were related to dangerous driving behaviour. The experience of fatigue could explained the underlying mechanism between these factors.

Junlong Guo,1,2 Lulu Yang,1,2 Yan Xu,1,2 Chenxi Zhang,1,2 Xian Luo,1,2 Shuai Liu,1– 3 Lihua Yao,4 Hanping Bai,4 Xiaofen Zong,4 Jihui Zhang,5 Zhongchun Liu,4 Bin Zhang1,2 1Department of Psychiatry, Nanfang Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 2Guangdong Mental Health Center, Guangdong Provincial People’s Hospital, Guangzhou, People’s Republic of China; 3Department of Psychiatry, The Third People’s Hospital of Qinghai Province, Xining, People’s Republic of China; 4Department of Psychiatry, Renmin Hospital of Wuhan University, Wuhan, People’s Republic of China; 5Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Southern Medical University, Guangzhou, Guangdong, People’s Republic of ChinaCorrespondence: Bin ZhangDepartment of Psychiatry, Nanfang Hospital, Southern Medical University, 1838 Guangzhou Avenue North, Guangzhou, 510515, People’s Republic of ChinaTel +86-20-62786731Email zhang73bin@hotmail.comZhongchun LiuDepartment of Psychiatry, Renmin Hospital of Wuhan University, 238 Jiefang Road, Wuhan, 430060, People’s Republic of ChinaEmail zcliu6@whu.edu.cnIntroduction: The prevalence rate and related factors of insomnia remained unknown after the COVID-19 epidemic had been under control. Therefore, we conducted this survey to investigate the prevalence rate and related factors of insomnia symptoms in the Chinese general public after the COVID-19 had been initially control.Methods: An online survey was conducted among Chinese citizens through the JD Health APP. The questionnaire was used for collecting demographic data and self-designed questions related to the COVID-19 outbreak. Insomnia Severity Index, Patient Health Questionnaire-9, Somatic Symptom Scale-8 and Impact of Events Scale-Revised were used for measuring psychological symptoms. To examine the associations of sociodemographic and psychological factors with insomnia symptoms, a binary logistic regression was used.Results: In total, there were 14,894 eligible participants, and 4601 (30.9%) participants were found to have insomnia symptoms. The regression model revealed that a higher risk of insomnia symptoms was associated with being over the age of 40 years, having history of psychiatric disorders, smoking, having infected friends or colleagues, having depressive or somatic symptoms, experiencing psychological distress and feeling estranged from family members. Meanwhile a lower risk of insomnia symptoms was associated with being female, having closer family relationships, not feeling alienated from others and being satisfied with the available information.Conclusion: In our study, 30.9% of the participants in the general public reported insomnia symptoms after the COVID-19 epidemic had been initially controlled. When providing precise interventions for insomnia, extra attention should be paid to the individuals who are male, elderly and smokers, and those with psychiatric disorder history, with infected friends or colleagues, with psychological symptoms and with poor social support.Keywords: insomnia symptoms, prevalence, COVID-19, under control, general public, China

Background Gastric adenocarcinoma with peritoneal carcinomatosis (PC) is therapy resistant and leads to poor survival. To study PC in depth, there is an urgent need to develop representative PC-derived cell lines and metastatic models to study molecular mechanisms of PC and for preclinical screening of new therapies. Methods PC cell lines were developed from patient-derived PC cells. The tumorigenicity and metastatic potential were investigated by subcutaneously (PDXs) and orthotopically. Karyotyping, whole-exome sequencing, RNA-sequencing, and functional studies were performed to molecularly define the cell lines and compare genomic and phenotypic features of PDX and donor PC cells. Results We established three PC cell lines (GA0518, GA0804, and GA0825) and characterized them in vitro. The doubling times were 22, 39, and 37 h for GA0518, GA0804, and GA0825, respectively. Expression of cancer stem cell markers (CD44, ALDH1, CD133 and YAP1) and activation of oncogenes varied among the cell lines. All three PC cell lines formed PDXs. Interestingly, all three PC cell lines formed tumors in the patient derived orthotopic (PDO) model and GA0518 cell line consistently produced PC in mice. Moreover, PDXs recapitulated transcriptomic and phenotypic features of the donor PC cells. Finally, these cell lines were suitable for preclinical testing of chemotherapy and target agents in vitro and in vivo. Conclusion We successfully established three patient-derived PC cell lines and an improved PDO model with high incidence of PC associated with malignant ascites. Thus, these cell lines and metastatic PDO model represent excellent resources for exploring metastatic mechanisms of PC in depth and for target drug screening and validation by interrogating GAC for translational studies.

Gastric cancer (GC) is one of the most common cancers globally, threatening global health. The deregulation of the Hippo signaling pathway has been discovered in GC and may be related to cancer development, proliferation, metastasis, and drug resistance. Yes-associated protein (YAP), as a downstream effector of the Hippo signaling pathway and a crucial co-transcription factor in the nucleus, is a promising and vital potential drug target for the treatment of GC. A series of drugs or compounds that inhibit YAP has been developed or confirmed. Therefore, this review will focus on summarizing the drugs and small-molecule inhibitors that have been reported to inhibit YAP and discuss the clinical prospects of YAP inhibitors in GC.

Background There are no effective antiviral therapies for coronavirus disease 2019 (COVID-19) at present. Although most patients with COVID-19 have a mild or moderate course of disease, up to 5%-10% of patients may have a serious and potentially life-threatening condition, indicating an urgent need for effective therapeutic drugs. The therapeutic effect of thymosin on COVID-19 has not been previously studied. In this paper, for the first time we report a case of thymosin treatment of COVID-19. Case summary A 51-year-old man with imported COVID-19 was admitted with definite symptoms of chest tightness, chest pain, and fatigue. The polymerase chain reaction results for severe acute respiratory syndrome coronavirus 2 were negative. The antibody test was positive, confirming the diagnosis of COVID-19. As many orally administered drugs were not well tolerated due to gastrointestinal symptoms, an emergency use of thymosin, a polypeptide consisting of 28 amino acids, was administered by injection. Finally, after the implementation of the treatment program, symptoms and lung imaging improved significantly. Conclusion In this case report, it is confirmed that thymosin may help alleviate the severity of COVID-19 symptoms.

Background The pancreatic immune‐related adverse event (irAE) is a rare but increasingly occurrence disease with limited knowledge, which was associated with the use of immune checkpoint inhibitors (ICIs). Methods In this case series study of pancreatic irAE patients, clinical and radiological manifestations are summarized. Baseline and post‐treatment fecal microbiota of immune‐related acute pancreatitis (irAP) patients were analyzed by the 16 s rDNA amplicon sequencing method. Results A total of six patients were enrolled into the study, and the onset of pancreatic irAEs occurred a median of 105 days after a median of 4.5 cycles with immune checkpoint inhibitors (ICIs). All patients had an effective response to ICIs. Abdominal pain was the main clinical manifestation. Serum amylase (sAMY) and lipase (sLIP) had dynamic changes parallel to clinical severity. Contrast‐enhanced computed tomography (CT) did not accurately reveal the level of inflammation. However, magnetic resonance imaging (MRI) was a sensitive imaging method which showed decreased and increased signal intensity of pancreatic parenchyma in T1‐weighted fat‐saturated and diffusion‐weighted imaging, respectively. Glucocorticoids were the main treatment with a rapid initial effect followed by a slow improvement. After reinitiation of ICI therapy, pancreatic irAEs either deteriorated, remained stable or the patient developed severe pancreatic β‐cell destruction without irAP recurrence. The baseline microbiota of irAP had low Bacteroidetes/Firmicutes ratio at phylum level, low relative abundance of Alistipes, Bacteroides and high Lachnospiraceae at genus level, compared to levels of pancreatic β‐cell destruction and post‐treatment of irAP. Conclusions Pancreatic irAE patients had corresponding abdominal pain and increase in sAMY/sLIP. MRI was found to be an ideal imaging modality. Treatment with glucocorticoids were the main approach. The microbiota showed relative changes at baseline and during treatment., Following onset and glucocorticoid treatment of severe immune‐related acute pancreatitis (irAP) (A), the serum amylase and lipase had parallel changes with the clinical conditions (B). The contrast‐enhanced CT did not accurately reveal the inflammation level; however, the MRI including T1‐weighted fat‐saturated, diffusion‐weighted image and magnetic resonance cholangiopancreatography clarified the radiological characteristics and dynamic changes of irAP (C). Correspondingly, the microbiota had relative changes at baseline and during treatment (D).

Background & Aims Liver fibrosis is a wound healing response that arises from various aetiologies. The intermediate filament protein Nestin has been reported to participate in maintaining tissue homeostasis during wound healing responses. However, little is known about the role Nestin plays in liver fibrosis. This study investigated the function and precise regulatory network of Nestin during liver fibrosis. Methods Nestin expression was assessed via immunostaining and quantitative real-time PCR (qPCR) in fibrotic/cirrhotic samples. The induction of Nestin expression by transforming growth factor beta (TGFβ)-Smad2/3 signalling was investigated through luciferase reporter assays. The functional role of Nestin in hepatic stellate cells (HSCs) was investigated by examining the pathway activity of profibrogenic TGFβ-Smad2/3 signalling and degradation of TGFβ receptor I (TβRI) after interfering with Nestin. The in vivo effects of knocking down Nestin were examined with an adeno-associated virus vector (serotype 6, AAV6) carrying short-hairpin RNA targeting Nestin in fibrotic mouse models. Results Nestin was mainly expressed in activated HSCs and increased with the progression of liver fibrosis. The profibrogenic pathway TGFβ-Smad2/3 induced Nestin expression directly. Knocking down Nestin promoted caveolin 1-mediated TβRI degradation, resulting in TGFβ-Smad2/3 pathway impairment and reduced fibrosis marker expression in HSCs. In AAV6-treated murine fibrotic models, knocking down Nestin resulted in decreased levels of inflammatory infiltration, hepatocellular damage, and a reduced degree of fibrosis. Conclusion The expression of Nestin in HSCs was induced by TGFβ and positively correlated with the degree of liver fibrosis. Knockdown of Nestin decreased activation of the TGFβ pathway and alleviated liver fibrosis both in vitro and in vivo. Our data demonstrate a novel role of Nestin in controlling HSC activation in liver fibrosis. Lay summary Liver fibrosis has various aetiologies but represents a common process in chronic liver diseases that is associated with high morbidity and mortality. Herein, we demonstrate that the intermediate filament protein Nestin plays an essential profibrogenic role in liver fibrosis by forming a positive feedback loop with the TGFβ-Smad2/3 pathway, providing a potential therapeutic target for the treatment of liver fibrosis.

Background Sputum cell‐free DNA (cfDNA) is a valuable surrogate sample for assessing EGFR‐sensitizing mutations in patients with advanced lung adenocarcinoma. Detecting EGFR exon 20 p.T790 M (p.T790 M) is much more challenging due to its limited availability in tumor tissues. Exploring sputum cfDNA as an alternative for liquid‐based sample type in detecting p.T790 M requires potential improvement in clinical practice. Methods A total of 34 patients with EGFR‐sensitive mutation‐positive lung adenocarcinoma and acquired resistance to the first generation of epidermal growth factor receptor‐tyrosine kinase inhibitors (EGFR‐TKIs) were enrolled. The sputum samples, and paired tumors and/or plasma samples were tested for p.T790 M mutation and concordance of p.T790 M status among the three sample types was analyzed. Results The overall concordance rate of p.T790 M mutation between sputum cfDNA and tumor tissue samples was 85.7%, with a sensitivity of 66.7% and a specificity of 100%. The sensitivity for detecting p.T790 M in sputum cfDNA was 100%, 66.7%, and 0% in the three sputum groups of malignant, satisfactory but no malignant cells, and unsatisfactory, respectively. The combined results of plasma cfDNA testing and sputum cfDNA testing further increased the sensitivity to 100% for p.T790 M detection in satisfactory but no malignant cells sputum group. Conclusion These findings revealed that cfDNA from malignant or satisfied but no malignant cells sputum is considered suitable for detecting p.T790 M mutation in patients with acquired resistance to first or second‐generation EGFR‐TKIs. The sputum cytological pathological evaluation‐guided sputum cfDNA testing assists in significantly improving the sensitivity of p.T790 M detection, bringing significant value for the maximal application of third‐generation EGFR‐TKIs in second‐line treatment., CfDNA from malignant or satisfied but no malignant cells sputum is suitable for detecting T790 M mutation in patients with acquired resistance to 1st or 2nd generation of EGFR‐TKIs. Sputum cytological pathological evaluation‐guided joint sputum and plasma cfDNA testing can significantly improve the sensitivity of T790 M detection, which brings significant value for maximal application of the third generation of EGFR‐TKI in second‐line treatment.

Long noncoding RNA (lnc RNA) is aberrant expressed in many kinds of tumors and may be concerned with the occurrence and progression of tumors. Lnc RNA MST1P2 is increased in cervical cancer (CC), but its mechanism in CC has not been clarified. In this study, RT-qPCR was employed to analyze Lnc MST1P2 and miR-133b expression. CCK8 and cell apoptosis assay detect the proliferation optical density (OD) value and apoptosis rate. Cell metastasis was evaluated by Wound-healing assay and Transwell assay. Dual-Luciferase assay analyzed the relationship between Lnc MST1P2 and miR-133b. In vivo experiment was performed by establishing xenograft animal model. We found that Lnc MST1P2 is obviously overexpression in CC tissues and cells. Si-Lnc MST1P2 obviously repressed cell growth, cell migration, and cell invasion in Hela and SIHA cells. Moreover, Si-Lnc MST1P2 suppressed CC tumorigenesis in vivo. Dual-Luciferase assay and RT-qPCR assay further proved that Lnc MST1P2 has a negative regulation to miR-133b. miR-133b up-regulation inhibited cell viability and metastasis of Hela and SIHA cells. miR-133b inhibition notably decreased the anti-cancer effect of si-Lnc MST1P2. LncRNA MST1P2 serves as a Cervical Cancer oncogene by sponging with miR-133b.

The coronavirus disease 2019 (COVID-19) pandemic has imposed both physical and psychological burdens on healthcare workers (HCWs). What is more, few studies have focused on the gender differences in mental health problems (MHPs) among HCWs during such an outbreak. Thus, the current study investigated the prevalence and gender differences of various MHPs among HCWs in China during the COVID-19 outbreak. This nationwide survey was conducted online from January 29 to February 3, 2020. General information was collected by questions about socio-demographics, work-related factors, and living situations. Depressive, anxiety, stress, and insomnia symptoms were assessed by the Patient Health Questionnaire-9, the Generalized Anxiety Disorder-7, the Impact of Event Scale-Revised, and the Insomnia Severity Index, respectively. Among the 2,198 contacted HCWs, 1,563 (71.1%) responded with valid data, of whom 1,293 (82.7%) were females. The prevalences of depressive, anxiety, stress, and insomnia symptoms in participants were 50.7%, 44.7%, 52.5%, and 36.1%, respectively. Female HCWs had significantly higher scores in all four scales (p

L-tert-leucine (L-Tle) is widely used as vital chiral intermediate for pharmaceuticals and as chiral auxiliarie for organocatalysis. L-Tle is generally prepared via the asymmetric reduction of trimethylpyruvate (TMP) catalyzed by NAD+-dependent leucine dehydrogenase (LeuDH). To improve the catalytic efficiency and coenzyme affinity of LeuDH from Bacillus cereus, mutation libraries constructed by error-prone PCR and iterative saturation mutation were screened by two kinds of high-throughput methods. Compared with the wild type, the affinity of the selected mutant E24V/E116V for TMP and NADH increased by 7.7- and 2.8-fold, respectively. And the kcat/Km of E24V/E116V on TMP was 5.4-fold higher than that of the wild type. A coupled reaction comprising LeuDH with glucose dehydrogenase of Bacillus amyloliquefaciens resulted in substrate inhibition at high TMP concentrations (0.5 M), which was overcome by batch-feeding of the TMP substrate. The total turnover number and specific space-time conversion of 0.57 M substrate increased to 11,400 and 22.8 mmol·h-1·L-1·g-1, respectively. KEY POINTS: • The constructed new high-throughput screening strategy takes into account the two indicators of catalytic efficiency and coenzyme affinity. • A more efficient leucine dehydrogenase (LeuDH) mutant (E24V/E116V) was identified. • E24V/E116V has potential for the industrial synthesis of L-tert-leucine.

In this work, the occurrence of 34 common antibiotic (15 sulfonamides and 19 quinolones) residues were evaluated in 236 ready-to-eat duck eggs (salted and preserved duck eggs) marketed through e-commerce stores by ultra-performance liquid chromatography coupled to tandem mass spectrometry, and subsequent dietary risk assessments for Chinese consumers were conducted. Among violated positives in duck egg samples, 11 analytes were found including sulfamethazine, sulfaquinoxaline, sulfamethoxazole, sulfadiazine, sulfamonomethoxine, ciprofloxacin, enrofloxacin, ofloxacin, flumequine, sarafloxacin, and nalidixic acid. A higher number of antibiotics were detected in salted duck eggs (five sulfonamides and six quinolones) than in preserved duck eggs (one sulfonamide and two quinolones). The maximum contamination of sulfonamides and quniolones was 448.0 µg/kg (sulfaquinoxaline) and 563.7 µg/kg (enrofloxacin) in salted duck eggs, respectively. Dietary exposure was evaluated through the estimated daily intake (EDI) of risky antibiotics (sulfamethazine, ciprofloxacin, and enrofloxacin) and hazard quotients (HQs). The results showed that EDIs and HQs were in the range of 0.0004 to 0.0099 µg/kg bw/day and 0.0009 to 0.1594%, respectively. The risk was low since HQs obtained were less than 100%. However, a special attention should be paid to ready-to-eat duck egg products high percentile consumers with the prosperity of e-commerce market in China. PRACTICAL APPLICATION: The present analytical method could be used for multiresidue determination of antibiotics in ready-to-eat duck eggs, and dietary risk assessments of risky antibiotics provided a support for the work of regulatory bodies to conduct surveillance programs regarding food safety evaluation of ready-to-eat foods.

Pathogens secrete a large number of effectors that manipulate host processes to create an environment conducive to pathogen colonization. However, the underlying mechanisms by which Plasmopara viticola effectors manipulate host plant cells remain largely unclear. In this study, we reported that RXLR31154, a P. viticola RXLR effector, was highly expressed during the early stages of P. viticola infection. In our study, stable expression of RXLR31154 in grapevine (Vitis vinifera) and Nicotiana benthamiana promoted leaf colonization by P. viticola and Phytophthora capsici, respectively. By yeast two-hybrid screening, the 23-kDa oxygen-evolving enhancer 2 (VpOEE2 or VpPsbP), encoded by the PsbP gene, in Vitis piasezkii accession Liuba-8 was identified as a host target of RXLR31154. Overexpression of VpPsbP enhanced susceptibility to P. viticola in grapevine and P. capsici in N. benthamiana, and silencing of NbPsbPs, the homologs of PsbP in N. benthamiana, reduced P. capcisi colonization, indicating that PsbP is a susceptibility factor. RXLR31154 and VpPsbP protein were co-localized in the chloroplast. Moreover, VpPsbP reduced H2 O2 accumulation and activated the 1 O2 signaling pathway in grapevine. RXLR31154 could stabilize PsbP. Together, our data revealed that RXLR31154 reduces H2 O2 accumulation and activates the 1 O2 signaling pathway through stabilizing PsbP, thereby promoting disease.

Objective This study aims to comparatively analyze the therapeutic efficacy upon multiple medication plans over Lopinavir/Ritonavir (LPV/r), Arbidol (ARB) and Methylprednisolone on patients with Coronavirus Disease 2019 (COVID‐19). Methods Totally 75 COVID‐19 patients admitted to The First Affiliated Hospital, Zhejiang University School of Medicine from January 22, 2020 to February 29, 2020 were recruited and grouped based on whether or not LPV/r and ARB were jointly used and whether or not Methylprednisolone was used. Indexes including body temperature, time for nucleic acid negative conversion, hospital stays and laboratory indexes were examined and compared. Results For all patients, there were no significant differences in the change of body temperature, the time for negative conversion and hospital stays whether LPV/r and ARB were jointly used or not. While for severe and critically severe patients, Methylprednisolone noticeably reduced the time for negative conversion. Meanwhile, the clinical efficacy was superior on patients receiving Methylprednisolone within 3 days upon admission, and the duration of hospital stays was much shorter when Methylprednisolone was given at a total dose of 0‐400 mg than a higher dose of >400 mg if all patients received a similar dose per day. Nonetheless, no significant changes across hepatic, renal and myocardial function indexes were observed. Conclusion LPV/r combined with ARB produced no noticeably better effect on COVID‐19 patients relative to the single agent treatment. Additionally, Methylprednisolone was efficient in severe and critically severe cases, and superior efficacy could be realized upon its early, appropriate and short‐term application. This article is protected by copyright. All rights reserved.

AIM: To investigate the clinical manifestations, diagnostic approaches, treatments, and outcomes of intraocular lymphoma. METHODS: In this retrospective study, 16 patients (28 eyes) with intraocular lymphoma were recruited in the Department of Ophthalmology, Peking Union Medical College Hospital, from 2004 to 2019. All patients underwent comprehensive ophthalmic examinations. Vitreous specimens of 13 patients were sent for cytopathology examination and other adjunctive diagnostic procedures. Three patients were diagnosed with intraocular lymphoma according to analysis of the histopathological results of systemic lymphoma by one clinician. Twenty-three eyes were treated with intravitreal administration of methotrexate, 4 eyes could not receive ocular treatment due to life-threatening lymphoma, and 1 eye did not require ocular treatment because the fundus lesions regressed after systematic chemotherapy. RESULTS: In 28 eyes, 25 eyes were diagnosed with vitreoretinal lymphoma, and 3 eyes were diagnosed with ciliary body lymphoma, all of which were non-Hodgkin diffuse large B cell lymphomas. The final visual acuity improved in 15 eyes (54%), remained unchanged in 5 eyes (18%), and decreased in 8 eyes (29%). Anterior segment inflammation disappeared or reduced in 8 and 5 eyes, respectively; and 15 eyes had no anterior segment reaction. Twenty eyes had mild vitreous opacity, 1 eye had mild vitritis, and 7 eyes had pars plana vitrectomy combined with silicone oil tamponade. Fundus lesions disappeared in 9 eyes and were relieved in 5 eyes; 4 eyes showed no changes, and the remaining 10 eyes' fundus were normal. CONCLUSION: The clinical manifestations of intraocular lymphoma are diverse, and the misdiagnosis rate is high. Cytopathological analysis of vitreous is one of the gold standards for the diagnosis. Immunohistochemistry, gene rearrangement and flow cytometric immunophenotypic analysis can improve the diagnostic rate. Ocular chemotherapy or radiotherapy regimens may preserve visual acuity, and a multidisciplinary team can provide individualized treatment for the patients.

Objectives The Bio-Rad SARS-CoV-2 ddPCR Kit (Bio-Rad Laboratories) was the first droplet digital polymerase chain reaction (ddPCR) assay to receive Food and Drug Administration (FDA) Emergency Use Authorization approval, but it has not been evaluated clinically. We describe the performance of ddPCR—in particular, its ability to confirm weak-positive severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) results. Methods We clinically validated the Bio-Rad Triplex Probe ddPCR Assay. The limit of detection was determined by using serial dilutions of SARS-CoV-2 RNA in an artificial viral envelope. The ddPCR assay was performed according to the manufacturer’s specifications on specimens confirmed to be positive (n = 48) or negative (n = 30) by an FDA-validated reverse transcription–polymerase chain reaction assay on the m2000 RealTime system (Abbott). Ten borderline positive cases were also evaluated. Results The limit of detection was 50 copies/mL (19 of 20 positive). Forty-seven specimens spanning a range of quantification cycles (2.9-25.9 cycle numbers) were positive by this assay (47 of 48; 97.9% positive precent agreement), and 30 negative samples were confirmed as negative (30 of 30; 100% negative percent agreement). Nine of 10 borderline cases were positive when tested in triplicate. Conclusions The ddPCR of SARS-CoV-2 is an accurate method, with superior sensitivity for viral RNA detection. It could provide definitive evaluation of borderline positive cases or suspected false-negative cases.

The prognosis of Artificial liver support system (ALSS) for hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) is hard to be expected, which results in multiple operations of ALSS and excessive consumption of plasma, increase in clinical cost. A total of 375 HBV-ACLF patients receiving ALSS treatment were randomly divided a train set and an independent test set. Logistic regression analysis was conducted and a decision tree was built based on 3-month survival as outcome. The ratio of total bilirubin before and after the first time of ALSS treatment was the most significant prognostic factor, we named it RPTB. Further, a decision tree based on the multivariate logistic regression model using CTP score and the RPTB was built, dividing patients into 3 main groups such as favorable prognosis group, moderate prognosis group and poor prognosis group. A clearly-presented and easily-understood decision tree was built with a good predictive value of prognosis in HBV-related ACLF patients after first-time ALSS treatment. It will help maximal the therapeutic value of ALSS treatment and may play an important role in organ allocation for liver transplantation in the future.

BACKGROUND: Recurrence of esophageal cancer (EC) after chemotherapy may mainly be explained by the existence of chemotherapy-resistant cells, and an effective drug against chemotherapy-resistant cells is highly sought. The aim of this study was to investigate the cytotoxicity of bispecific antibody solitomab combined with γ δ T cells on Eca109 cell spheres. METHODS: We cultured Eca109 cell spheres in serum-free medium, and the morphological differences between wild-type Eca109 cells and Eca109 cell spheres were compared by microscope and flow cytometry. Different concentrations of nanoparticle albumin-bound paclitaxel (Nab-PTX) and cisplatin were used to treat the two groups of cells and compare their drug resistance. Flow cytometry was then used to detect the expression level of epithelial cell adhesion molecule (EpCAM) and the cytotoxicity of γ δ T cells combined with bispecific antibody solitomab on the two groups. RESULTS: Flow cytometry analysis showed that Eca109 cell spheres were smaller in size and had less cytoplasmic granules and CCK-8 assay showed that the viability of Eca109 cell spheres treated with different concentrations of Nab-PTX and cisplatin was significantly higher than that of wild-type Eca109 cells (P

Background Fermented foods, with a history as long as the human civilization, form an indispensable constituent in our daily life. However, most fermented foods are produced by spontaneous fermentation, and the fermentation processes are still uncontrolled. To ensure consistent food quality, it is of paramount importance to understand and control the spontaneous food fermentations. Scope and approach In spontaneous food fermentations, metabolic activity of a succession of complex microbiota results in desired flavour that is the key criterion to decide consumers’ preference. Therefore, flavour compound formation by microbial metabolism can be used as the control target in spontaneous food fermentations. However, relatively little is known about the complexity of the microbiota associated with the flavour compound formation. Therefore, in this review by using Chinese liquor as a model system, we present key biotechnological aspects of the microbiota crucial for flavour of fermented foods, including the driving forces for the microbiota succession, flavour compound formation, and the regulation of flavour compound formation. Key findings and conclusions Core microbiota, associated with flavour compound formation, can be identified and eventually used to construct a synthetic (i.e. designed) microbiota. Meanwhile, key environmental factors, affecting the core microbiota, can also be identified and controlled to regulate the synthetic microbiota. Furthermore, modelling can be used to predict, optimize and control the flavour compound formation by the synthetic microbiota, so that such spontaneous food fermentations can become controllable, with the ultimate goal to monitor, control, and improve the quality, productivity and safety of fermented foods.

At present, there is no method to predict or monitor patients with AMI, and there is no specific treatment method. In order to improve the analysis of clinical influencing factors of acute myocardial infarction, based on the machine learning algorithm, this paper uses the K-means algorithm to carry out multifactor analysis and constructs a hybrid model combined with the ART2 network. Moreover, this paper simulates and analyzes the model training process and builds a system structure model based on the KNN algorithm. After constructing the model system, this paper studies the clinical influencing factors of acute myocardial infarction and combines mathematical statistics and factor analysis to carry out statistical analysis of test results. The research results show that the system model constructed in this paper has a certain effect in the clinical analysis of acute myocardial infarction.

Asarum is a traditional medicine and has been widely used as remedies for inflammatory diseases, toothache, headache, local anesthesia, and aphthous stomatitis in China, Japan, and Korea. Our previous research found that safrole and methyl eugenol were vital compounds that contribute to distinguish the different species and raw Asarum and its processed products apart. The pharmacokinetics of safrole and methyl eugenol after oral administration of Asarum extract has not been reported yet. In this study, a rapid and simple gas chromatography–mass spectroscopy (GC–MS) method that has a complete run time of only 4.5 min was developed and validated for the simultaneous determination and pharmacokinetic study of safrole and methyl eugenol in rat plasma after administration of Asarum extracts. The chromatographic separation was realized on a DB-17 column ( 30 m × 0.25 mm × 0.25 μ m ). And detection was carried out under selected ion monitoring (SIM) mode. Plasma samples were pretreated by n -hexane. The pharmacokinetic parameters provided by this study will be beneficial for further developments and clinical applications of Asarum.

Nodal-skip metastasis (NSM) is found in esophageal squamous cell carcinoma (ESCC), but its prognostic role is controversial. This study aimed to investigate the prognostic value of NSM for thoracic ESCC patients. Categorization of NSM was according to the N groupings of Japan Esophagus Society (JES) staging system, which is dependent on tumor location. Using the Kaplan–Meier method and Cox-regression analysis, this study retrospectively analyzed the overall survival (OS) for 2325 ESCC patients after radical esophagectomy at three high-volume esophageal cancer centers. Predictive models also were constructed. The overall NSM rate was 20% (229/1141): 37.4% in the in upper, 12.9% in the middle, and 22.2% in the lower thoracic ESCC. The patients with NSM always had a better prognosis than those without NSM. Furthermore, NSM was an independent prognostic factor for thoracic ESCC patients (hazard ratio [HR], 0.633; 95% confidence interval [CI], 0.499–0.803; P

Background: Achilles tendon (AT) defects often occur in traumatic and chronic injuries. Currently, no graft can satisfactorily regenerate parallel tendinous tissue at the defect site to completely restore AT function. Purpose: To develop a cell-free functional graft by tethering bone morphogenetic protein 12 (BMP-12) on a book-shaped decellularized tendon matrix (BDTM) and to determine whether this graft is more beneficial for AT defect healing than an autograft. Study Design: Controlled laboratory study. Methods: Canine patellar tendon was sectioned into a book shape and decellularized to fabricate a BDTM. The collagen-binding domain (CBD) was fused into the N-terminus of BMP-12 to synthesize a recombinant BMP-12 (CBD-BMP-12), which was tethered to the BDTM to prepare a cell-free functional graft (CBD-BMP-12/BDTM). After its tensile resistance, tenogenic inducibility, and BMP-12 release dynamics were evaluated, the efficacy of the graft for tendon regeneration was determined in a rat model. A total of 140 mature male Sprague-Dawley rats underwent AT tenotomy. The defect was reconstructed with reversed AT (autograft group), native BMP-12 tethered to an intact decellularized tendon matrix (IDTM; NAT-BMP-12/IDTM group), native BMP-12 tethered to a BDTM (NAT-BMP-12/BDTM group), CBD-BMP-12 tethered on an IDTM (CBD-BMP-12/IDTM group), and CBD-BMP-12 tethered on a BDTM (CBD-BMP-12/BDTM group). The rats were sacrificed 4 or 8 weeks after surgery to harvest AT specimens. Six specimens from each group at each time point were used for histological evaluation; the remaining 8 specimens were used for biomechanical testing. Results: In vitro CBD-BMP-12/BDTM was noncytotoxic, showed high biomimetics with native tendons, was suitable for cell adhesion and growth, and had superior tenogenic inducibility. In vivo the defective AT in the CBD-BMP-12/BDTM group regenerated more naturally than in the other groups, as indicated by more spindle-shaped fibroblasts embedded in a matrix of parallel fibers. The biomechanical properties of the regenerated AT in the CBD-BMP-12/BDTM group also increased more significantly than in the other groups. Conclusion: CBD-BMP-12/BDTM is more beneficial than autograft for healing AT defects in a rat model. Clinical Relevance: The findings of this study demonstrate that CBD-BMP-12/BDTM can serve as a practical graft for reconstructing AT defects.

Dysfunction of the sleep-wake transition is considered to be associated with the pathology of patients with primary insomnia (PI). Previous animal study had reported that brain circuits between the striatum and cortex can regulate sleep-wake transitions. So far, few studies have systematically explored the structural connectivity of the striatum-centered circuits and their potential roles in patients with PI. In this study, we chosen the striatum as the seed and 10 priori target regions as masks to assess the structural connectivity by using seed-based classification with a diffusion tensor imaging (DTI) probabilistic tractography method. Track strengths of the striatum-centered circuits were compared between 22 patients with PI (41.27 ± 9.21 years) and 30 healthy controls (HC) (35.2 ± 8.14 years). Pittsburgh Sleep Quality Index (PSQI) was used to measure the sleep quality in all participants. Lower track strengths (left striatum- anterior cingulate cortex (ACC), left striatum- dorsal anterior cingulate cortex (dACC), left striatum-Hippocampus, and right striatum-Hippocampus) were observed in patients with PI compared to HC. Additionally, the lower track strengths of brain circuits mentioned above were negatively correlated with PSQI. Taken together, our findings revealed the lower tract strength of frontostriatal circuits in patients with PI and HC, which provided the implications of the system-level structural connections of frontostriatal circuits in the pathology of PI. We suggested that the track strengths of the frontostriatal circuits calculated from DTI can be the potential neuroimaging biomarkers of the sleep quality in patients with PI.

Purpose To investigate the change in muscle volume around the hip in patients with femoroacetabular impingement (FAI) after arthroscopy and evaluate other factors related to muscle change. Methods We performed a retrospective review of magnetic resonance imaging data of patients with FAI who underwent hip arthroscopy. Magnetic resonance imaging was obtained pre- and postoperatively. The cross-sectional area (CSA) of muscles were determined on axial images. The Wilcoxon signed-rank test was used to determine the differences between pre- and postoperative hip muscle CSA. The correlations of change in muscle CSA with age, sex, body mass index, pain level, preoperative symptom duration, follow-up time, and multiple validated patient-reported outcomes were also analyzed with a Spearman rank correlation test. Results Fifty-one patients with a mean age of 36.5 ± 5.6 years were included and analyzed. The follow-up was 26.6 ± 0.5 months (range, 24-40 months), and 27 (52.9%) were women. Patients with FAI showed increased hip muscle CSA of gluteus maximus (P = .002) and gluteus minimus (P = .001). Post- compared with preoperative, the value for the change in medius CSA was underpowered, and no differences in other hip muscle CSAs were observed. The increased muscle CSA of the gluteus maximus was significantly correlated with the improvement of modified Harris Hip Score (ρ = 0.404; P = .003). The increased muscle CSA of the gluteus minimus was significantly correlated with the improvement of pain Visual Analog Scale (ρ = 0.452; P = .001). Age, body mass index, sex, symptom duration, and follow-up time were not significantly correlated with change in muscle CSA. Conclusions Patients with FAI have a significantly increased postoperative muscle CSA of the gluteus maximus (7.8%) and the gluteus minimus (11.6%) compared with preoperative values. The increased muscle CSA of the gluteus maximus and gluteus minimus was significantly correlated with improvement in modified Harris Hip Score and pain Visual Analog Scale, respectively. The increase of muscle volume may be associated with the improvement of subjective function and pain relief. Levels of Evidence Level IV, therapeutic case series.

Interrupted axons that fail to regenerate mainly cause poor recovery after spinal cord injury (SCI). How neurons epigenetically respond to injury determines the intrinsic growth ability of axons. However, the mechanism underlying epigenetic regulation of axonal regeneration post-SCI remains largely unknown. In this study, we elucidated the role of the epigenetic regulatory network involving ubiquitously transcribed tetratricopeptide repeat on chromosome X (UTX)/microRNA-24 (miR-24)/NeuroD1 in axonal regeneration and functional recovery in mice following SCI. Our results showed that UTX was significantly increased post-SCI and repressed axonal regeneration in vitro. However, downregulation of UTX remarkably promoted axonal regeneration. Furthermore, miR-24 was increased post-SCI and positively regulated by UTX. miR-24 also inhibited axonal regeneration. Chromatin immunoprecipitation (ChIP) indicated that UTX binds to the miR-24 promoter and regulates miR-24 expression. Genome sequencing and bioinformatics analysis suggested that NeuroD1 is a potential downstream target of UTX/miR-24. A dual-luciferase reporter assay indicated that miR-24 binds to NeuroD1; moreover, it represses axonal regeneration by negatively regulating the expression of NeuroD1 via modulation of microtubule stability. UTX deletion in vivo prominently promoted axonal regeneration and improved functional recovery post-SCI, and silencing NeuroD1 restored UTX function. Our findings indicate that UTX could be a potential target in SCI., Graphical Abstract, Guo et al. described an epigenetic regulatory mechanism of axon regeneration after spinal cord injury (SCI). UTX and miR-24 form an epigenetic complex to regulate expression of NeuroD1, which further influences microtubule stability and axon regeneration after SCI. This opens new possibilities for therapeutic strategies of SCI.

Nicotinamide adenine dinucleotide (NAD+) is an essential coenzyme involved in numerous physiological processes. As an attractive product in the industrial field, NAD+ also plays an important role in oxidoreductase-catalyzed reactions, drug synthesis, and the treatment of diseases, such as dementia, diabetes, and vascular dysfunction. Currently, although the biotechnology to construct NAD+-overproducing strains has been developed, limited regulation and low productivity still hamper its use on large scales. Here, we describe multi-strategy metabolic engineering to address the NAD+-production bottleneck in E. coli. First, blocking the degradation pathway of NAD(H) increased the accumulation of NAD+ by 39%. Second, key enzymes involved in the Preiss-Handler pathway of NAD+ synthesis were overexpressed and led to a 221% increase in the NAD+ concentration. Third, the PRPP synthesis module and Preiss-Handler pathway were combined to strengthen the precursors supply, which resulted in enhancement of NAD+ content by 520%. Fourth, increasing the ATP content led to an increase in the concentration of NAD+ by 170%. Finally, with the combination of all above strategies, a strain with a high yield of NAD+ was constructed, with the intracellular NAD+ concentration reaching 26.9 μmol/g DCW, which was 834% that of the parent strain. This study presents an efficient design of an NAD+-producing strain through global regulation metabolic engineering.

Yu-Hong Zhao, Yan Xu Operating Room, Linyi Central Hospital, Linyi, Shangdong, 276400, People’s Republic of ChinaCorrespondence: Yan XuOperating Room, Linyi Central Hospital, No. 17 Health Road, Yicheng Street, Yishui County, Linyi, Shangdong, 276400, People’s Republic of ChinaTel +86-15964811679Email 623535870@qq.comObjective: The aim of our current study is to explore the effect of comprehensive nursing based on cognitive behavior on the psychological function of glioma patients.Methods: A total of 108 patients with glioma were randomly divided into intervention group (n = 54) and control group (n = 54). Patients in the control group were given routine nursing for 3 months, while patients in the intervention group were given comprehensive nursing based on cognitive behavior for 3 months on the basis of control group. Parameters such as scores of simplified Fear of Progression Questionnaire-Short Form (FoP-Q-SF), Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS) as well as cancer-related fatigue and quality of life (QOL) were compared before and after the intervention.Results: No significant difference was observed in the scores of FoP-Q-SF, SAS, SDS, cancer-related fatigue and quality of life between the two groups before intervention (P > 0.05). Conversely, after the intervention, the scores of FoP-Q-SF, SAS, SDS and cancer-related fatigue in the intervention group were significantly lower than those in the control group, and the scores of life quality in the intervention group were significantly higher than those in the control group (all P values < 0.05).Conclusion: Our findings provide evidence that comprehensive nursing based on cognitive behavior can effectively improve the negative psychological state of patients such as anxiety and depression and cancer-related fatigue with improved life quality for patients harboring glioma.Keywords: comprehensive nursing based on cognitive behavior, glioma, psychological function

VPS33B is reported to be a tumor suppressor in hepatocellular carcinoma, nasopharyngeal carcinoma, colon cancer, and lung adenocarcinoma. Here, we observed that reduced VPS33B protein level was an unfavorable factor that promoted the pathogenesis of non-small cell lung cancer (NSCLC) in clinical specimens. We achieved lentivirus-mediated stable overexpression of VPS33B in NSCLC cells. Increased VPS33B reduced cell cycle transition and cell proliferation of NSCLC cells in vivo and in vitro. Knocking down VPS33B restored cell growth. Mechanism analysis indicated that miR-192-3p was induced by VPS33B and acted as a tumor suppressor of cell growth in NSCLC. Further, c-Myc or p53 was identified as a transcription factor that bound to the miR-192-3p promoter and regulated its expression. miR-192-3p directly targeted cell cycle-promoted factor CCNB1 and suppressed NSCLC cell growth. VPS33B modulated c-Myc/p53/miR-192-3p signaling to target CCNB1 by reducing activation of the Ras/ERK pathway. Our study reveals a novel molecular basis for VPS33B as a tumor suppressor to participate in the pathogenesis of NSCLC., Graphical Abstract, The dysfunction of a large number of genes participates in the pathogenesis of NSCLC. Liu and colleagues demonstrated that VPS33B acts as a tumor suppressor to halt NSCLC pathogenesis and elucidated the underlying mechanism by which VPS33B regulates c-Myc/p53/miR-192-3p to target CCNB1 via inactivating the Ras/ERK signal.

Small RNAs play a major role in the post-transcriptional regulation of gene expression in eukaryotes. Despite the evolutionary importance of streptophyte algae, knowledge on small RNAs in this group of green algae is almost non-existent. We used genome and transcriptome data of 34 algal and plant species, and performed genome-wide analyses of small RNA (miRNA & siRNA) biosynthetic and degradation pathways. The results suggest that Viridiplantae started to evolve plant-like miRNA biogenesis and degradation after the divergence of the Mesostigmatophyceae in the streptophyte algae. We identified two major evolutionary transitions in small RNA metabolism in streptophyte algae; during the first transition, the origin of DCL-New, DCL1, AGO1/5/10 and AGO4/6/9 in the last common ancestor of Klebsormidiophyceae and all other streptophytes could be linked to abiotic stress responses and evolution of multicellularity in streptophytes. During the second transition, the evolution of DCL 2,3,4, and AGO 2,3,7 as well as DRB1 in the last common ancestor of Zygnematophyceae and embryophytes, suggests their possible contribution to pathogen defense and antibacterial immunity. Overall, the origin and diversification of DICER and AGO along with several other small RNA pathway-related genes among streptophyte algae suggested progressive adaptations of streptophyte algae during evolution to a subaerial environment., Wang, Liang et al. conduct a genome-wide investigation into the origin of small RNA pathway-related genes in Viridiplantae. Their findings suggest that streptophyte algae progressively adapted to a subaerial environment through generation of these pathways.

To investigate the effects of RNA-binding proteins cold-inducible RNA binding protein (CIRP) and human antigen R (HuR) on expression of Claudin1 and mucosal barrier function in ulcerative colitis (UC). The clinical specimens of UC patients and healthy volunteers were collected. In the clinical experiments, the expressions of CIRP, Claudin1, and HuR, along with their correlations in tissues of UC patients were analyzed by qRT-PCR, Western blot and Pearson correlation coefficient, respectively. The chi-square test was utilized to assess the relevance between CIRP/HuR/Claudin1 level and clinicopathological characteristics of UC patients. The in vitro and in vivo models of UC were established by lipopolysaccharide treatment or dextran sulfate sodium injection. For cell experiments, after loss- and gain-of-function, the roles of CIRP or HuR in the apoptosis and proliferation of enterocytes were examined by flow cytometry and CCK-8 assay. The intestinal epithelial barrier function was inspected after determination on transepithelial electrical resistance value, horseradish peroxidase permeability and expressions of tight junction proteins (Occludin, ZO-1, and JAM-1). The relationship between HuR, CIRP, and Claudin1 was performed by RNA immunoprecipitation and dual-luciferase reporter gene assay. For in vivo experiments, the disease activity index score, weight loss and colon length of mice were assessed to observe the effect of CIRP or HuR on the UC mouse models. Histological analysis of colon tissues was conducted by H&E staining. FITC-dextran tracking was applied to inspect the intestinal mucosal barrier function of UC mouse models. In this study, high expression of CIRP and low expressions of HuR and Claudin1 were observed in patients, cells and mouse models of UC. The expressions of CIRP, HuR, and Claudin1 were correlated with the severity of patients with UC. There was a negative correlation between CIRP and Claudin1, and as a positive correlation between HuR and Claudin1. Claudin1 can be suppressed by CIRP, while enhanced by HuR. HuR and CIRP can competitively bind to Claudin1. HuR upregulation or CIRP downregulation promoted proliferation, suppressed apoptosis and ameliorated the damage of the barrier function in enterocytes. The in vivo experiments verified that the ameliorated damage of the intestinal mucosal barrier function in UC mice occurred with HuR overexpression or CIRP knockdown. CIRP and HuR confer pivotal effect on the intestinal mucosal barrier function of UC through competitively binding to Claudin1 mRNA.

INTRODUCTION Nicotiflorin is the main characteristic component of Nymphaea candida, which is a natural product that reportedly ameliorates acute injury of the liver and cerebral cortex, but the effect of nicotiflorin on acute kidney injury (AKI) remains unknown. This study aimed to investigate the effects of nicotiflorin on ischaemia/reperfusion (I/R) AKI and the associated mechanisms. METHODS We performed both (a) in vivo experiments with C57BL/6 mice with bilateral renal pedicles clamped for 45 minutes and (b) in vitro experiments with human kidney epithelial cells (HK-2) exposed to hypoxia/reoxygenation to mimic I/R injury to study the role of nicotiflorin in AKI. RESULTS In vivo, nicotiflorin administration exerted protective effects on renal injury, as demonstrated by reductions in the levels of caspase3 and Bad (P

The PAX3-FOXO1 fusion protein is the key oncogenic driver in fusion positive rhabdomyosarcoma (FP-RMS), an aggressive soft tissue malignancy with a particularly poor prognosis. Identifying key downstream targets of PAX3-FOXO1 will provide new therapeutic opportunities for treatment of FP-RMS. Herein, we demonstrate that Forkhead Box F1 (FOXF1) transcription factor is uniquely expressed in FP-RMS and is required for FP-RMS tumorigenesis. The PAX3-FOXO1 directly binds to FOXF1 enhancers and induces FOXF1 gene expression. CRISPR/Cas9 mediated inactivation of either FOXF1 coding sequence or FOXF1 enhancers suppresses FP-RMS tumorigenesis even in the presence of PAX3-FOXO1 oncogene. Knockdown or genetic knockout of FOXF1 induces myogenic differentiation in PAX3-FOXO1-positive FP-RMS. Over-expression of FOXF1 decreases myogenic differentiation in primary human myoblasts. In FP-RMS tumor cells, FOXF1 protein binds chromatin near enhancers associated with FP-RMS gene signature. FOXF1 cooperates with PAX3-FOXO1 and E-box transcription factors MYOD1 and MYOG to regulate FP-RMS-specific gene expression. Altogether, FOXF1 functions downstream of PAX3-FOXO1 to promote FP-RMS tumorigenesis.

Rationale: The regeneration and replacement of lung cells or tissues from induced pluripotent stem cell- or embryonic stem cell-derived cells represent future therapies for life-threatening pulmonary disorders but are limited by technical challenges to produce highly differentiated cells able to maintain lung function. Functional lung tissue-containing airways, alveoli, vasculature, and stroma have never been produced via directed differentiation of embryonic stem cells (ESCs) or induced pluripotent stem cells. We sought to produce all tissue components of the lung from bronchi to alveoli by embryo complementation.Objectives: To determine whether ESCs are capable of generating lung tissue in Nkx2-1-/- mouse embryos with lung agenesis.Methods: Blastocyst complementation was used to produce chimeras from normal mouse ESCs and Nkx2-1-/- embryos, which lack pulmonary tissues. Nkx2-1-/- chimeras were examined using immunostaining, transmission electronic microscopy, fluorescence-activated cell sorter analysis, and single-cell RNA sequencing.Measurements and Main Results: Although peripheral pulmonary and thyroid tissues are entirely lacking in Nkx2-1 gene-deleted embryos, pulmonary and thyroid structures in Nkx2-1-/- chimeras were restored after ESC complementation. Respiratory epithelial cell lineages in restored lungs of Nkx2-1-/- chimeras were derived almost entirely from ESCs, whereas endothelial, immune, and stromal cells were mosaic. ESC-derived cells from multiple respiratory cell lineages were highly differentiated and indistinguishable from endogenous cells based on morphology, ultrastructure, gene expression signatures, and cell surface proteins used to identify cell types by fluorescence-activated cell sorter.Conclusions: Lung and thyroid tissues were generated in vivo from ESCs by blastocyst complementation. Nkx2-1-/- chimeras can be used as "bioreactors" for in vivo differentiation and functional studies of ESC-derived progenitor cells.

Objective This study investigates variations in night, day, and total sleep trajectories across infancy and childhood in Asian children. Participants Participants consisted of a subset of 901 children, within the Growing Up in Singapore Towards healthy Outcomes cohort, which recruited 1247 pregnant women between June 2009 and September 2010. Design We used a novel conditional probabilistic trajectory model: a probabilistic model for mixture distribution, allowing different trajectory curves and model variances among groups to cluster longitudinal observations. Longitudinal sleep duration data for the trajectory analyses were collected from caregiver-reported questionnaires at 3, 6, 9, 12, 18, 24, and 54 months. Results We found 3 patterns of night sleep trajectories (n = 356): long consistent (31%), moderate consistent (41%), and short variable (28%); and 4 patterns of day sleep trajectories (n = 347): long variable (21%), long consistent (20%), moderate consistent (34%), and short consistent (25%). We also identified 4 patterns of total sleep trajectories (n = 345): long variable (19%), long consistent (26%), moderate consistent (28%), and short variable (27%). Short, moderate, and long trajectories differed significantly in duration. Children with consistent trajectories also displayed sleep patterns that were significantly more representative of typical developmental sleep patterns than children with variable trajectories. Conclusions This is the first study to describe multiple sleep trajectories in Singaporean children and identify between-individual variability within the trajectory groups. Compared to predominantly Caucasian samples, night/total sleep trajectories were generally shorter, while day sleep trajectories were longer. Future studies should investigate how these variations are linked to different developmental outcomes.

This study evaluated the effects of mitral regurgitation (MR) on cardiac structure and function in left ventricular noncompaction (LVNC) patients. The clinical and cardiovascular magnetic resonance (CMR) data for 182 patients with noncompaction or hypertrabeculation from three institutes were retrospectively included. We analyzed the difference in left ventricular geometry, cardiac function between LVNC patients with and without MR. The results showed that patients with MR had a worse New York Heart Association (NYHA) class and a higher incidence of arrhythmia (P

High levels of impulsivity are a risk factor for the initiation of heroin use and a core behavioral characteristic of heroin dependence. Impulsivity also contributes to the maintenance of drug use and hinders effective therapy. Here we sought to identify neuroimaging markers of impulsivity in heroin-dependent individuals (HDI), with a focus on the nucleus accumbens (NAc), a key region implicated in impulsivity and drug addiction generally. Volume and resting-state functional connectivity (RSFC) differences of the bilateral NAc were investigated between 21 HDI and 21 age-, gender-, nicotine-, alcohol-matched healthy controls (HC). The neuroimaging results were then correlated with the Barratt Impulsivity Scales (BIS-11). Higher motor impulsivity (t = 2.347, p = 0.0253) and larger right NAc volume (F (1,38) = 4.719, p = 0.036) was observed in HDI. The right NAc volume was positively correlated with BIS total (r = 0.6196, p = 0.0239) /motor (r = 0.5921, p = 0.0330) scores in HC and BIS motor (r = 0.5145, p = 0.0170) score in HDI. A negative correlation was found between RSFC of the right NAc-bilateral superior frontal gyrus (SFG) and motor impulsivity in HDI (left: r=-0.6537, p = 0.0013; right: r=-0.6167, p = 0.0029) and HC (left: r=-0.6490,p = 0.0164; right: r=-0.6993, p = 0.0078). We aimed to reveal novel multimodality neuroimaging biomarkers of the higher impulsivity in HDI by focusing on the NAc and corresponding functional circuits. Higher motor impulsivity was observed in HDI. Furthermore, the volume of the right NAc and the RSFC strength of right NAc-SFG could be neuroimaging biomarkers for the severity of impulsivity in HDI. These potential biomarkers could be a target for novel treatments in HDI.

Di Yang,1,* Lin-xia Shen,1,2,* Ri-feng Chen,1,* Yu Fu,1 Hong-yan Xu,1 Li-na Zhang,1 Dong-hua Liu1,3 1Department of Dermatology, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, People’s Republic of China; 2Department of Dermatology and Venereology, Huashan Hospital, Fudan University, Shanghai, 200040, People’s Republic of China; 3Guangxi Key Laboratory of AIDS Prevention and Treatment, School of Public Health, Nanning, 530021, People’s Republic of China*These authors contributed equally to this workCorrespondence: Dong-hua LiuDepartment of Dermatology, The First Affiliated Hospital of Guangxi Medical University, No.6, Shuangyong Road, Qingxiu District, Nanning, Guangxi Province, 530021, People’s Republic of ChinaEmail ldhgxmu@163.comBackground: Talaromyces marneffei (T. marneffei) is a destructive opportunistic dimorphic fungal which can cause lethiferous Talaromycosis, but the clearance of T. marneffei mainly depends on the innate immune response.Objective: To investigate whether T. marneffei can inhibit the expression of CD86 in THP-1 cells after infection and discuss the potential mechanisms.Methods: Western blot and immunoelectron microscopy were used to detect the CD86 expression on T. marneffei cultured on BHI medium at 37°C. Western blot, enzyme-linked immunoassay and immunofluorescence were used to detect the change of CD86 expression on macrophages incubating with T. marneffei. Enzyme-linked immunoassay was used to detect the content of CD86 in supernatant in the co-culture system. Immunohistochemistry and immunoelectron microscopy were used to detect the expression of CD86 on T. marneffei incubating with macrophages.Results: T. marneffei did not express CD86 when cultured separately at 37°C detected by Western blot and immunoelectron microscopy, but it did express CD86 when incubated with macrophages detected by immunohistochemistry and immunoelectron microscopy. The CD86 expression of macrophages significantly decreased at 72 hours when infected with T. marneffei while the content of CD86 in supernatant significantly increased at 72 hours compared with the control group which were detected by Western blot, enzyme-linked immunoassay and immunofluorescence.Conclusion: 1) After T. marneffei infection, CD86 expression on THP-1 decreased, and with the progression of infection, insufficient polarization of M1 macrophages gradually appeared; 2) T. marneffei may adsorb or uptake CD86 in supernatant produced by macrophages during the contact with THP-1 cells, thus leading to the consumption of CD86 in macrophages.Keywords: Talaromyces marneffei, T. marneffei, macrophages, CD86, THP-1, immunoelectron microscopy

Background Von Willebrand factor (VWF) contains a number of free thiols, the majority of which are located in its C-domains, and these have been shown to alter VWF function, However, the impact of free thiols on function following acute exposure of VWF to collagen under high and pathological shear stress has not been determined. Methods VWF free thiols were blocked with N-ethylmaleimide and flow assays performed under high and pathological shear rates to determine the impact on platelet capture and collagen binding function. Atomic force microscopy (AFM) was used to probe the interaction of VWF with collagen and molecular simulations conducted to determine the effect of free thiols on the flexibility of the VWF-C4 domain. Results Blockade of VWF free thiols reduced VWF-mediated platelet capture to collagen in a shear-dependent manner, with platelet capture virtually abolished above 5000 s-1 and in regions of stenosis in microfluidic channels. Direct visualization of VWF fibers formed under extreme pathological shear rates and analysis of collagen-bound VWF attributed the effect to altered binding of VWF to collagen. AFM measurements showed that thiol-blockade reduced the lifetime and strength of the VWF-collagen bond. Pulling simulations of the VWF-C4 domain demonstrated that with one or two reduced disulphide bonds the C4 domain has increased flexibility and the propensity to undergo free-thiol exchange. Conclusions We conclude that free thiols in the C-domains of VWF enhance the flexibility of the molecule and enable it to withstand high shear forces following collagen binding, demonstrating a previously unrecognized role for VWF free thiols.

Our study aimed to comprehensively explore efficient prognostic indicators in idiopathic dilated cardiomyopathy (IDCM) patients with reduced left ventricular ejection fraction (LVEF40%).Prognostic value of cardiac magnetic resonance(CMR) parameters for IDCM have been inconsistent.126 IDCM patients with reduced LVEF (40%) were retrospectively enrolled. Cardiac function parameters, myocardial strain indices and myocardial fibrosis were evaluated. Laboratory data also were analyzed. The endpoint was a combination of major adverse cardiac events (MACEs), including cardiac death, heart transplantation, and rehospitalization. Prognostic value was evaluated by the Kaplan-Meier method and Cox regression.During a median follow-up of 31 months, 44 patients experienced MACEs, including 9 deaths, 1 heart transplantation, and 34 rehospitalizations due to heart failure. Univariate and multivariate Cox analyses showed that cardiac function and myocardial strain indexes were not associated with the prognosis of IDCM (all p0.05). NT-proBNP (HR 1.5, 95%CI: 1.053 to 2.137), Late‑gadolinium enhancement(LGE) mass (HR 1.022, 95%CI: 1.005 to 1.038), and LGE mass/left ventricle mass were significant predictors (HR 1.027, 95%CI: 1.007 to 1.046) for MACEs, all p 0.05. Besides, poorest prognosis was observed in IDCM patients with positive LGE combined with NT-proBNP (log-rank = 27.261, p ≤ 0.001).NT-proBNP and extent of LGE were reliable predictors in IDCM patients with reduced LVEF. Additionally, presence of LGE combined with NT-proBNP showed the strongest prognostic value in IDCM with reduced LVEF. Myocardial strain parameters seemed to have no prognostic value in IDCM patients with reduced LVEF.

Background Positron emission tomography/computed tomography (PET/CT) has been recognized for diagnosing and staging lung cancer, but the prognostic value of standardized uptake value (SUV) on 18 F-FDG PET/CT imaging in patients with advanced non-small cell lung cancer (NSCLC) remains controversial. Methods We performed a retrospective analysis of patients with advanced NSCLC who had undergone 18 F-FDG PET/CT before systemic treatment between June 2012 and June 2016. The relationship between the maximum SUV (SUVmax) of the pulmonary lesion and lesion size was evaluated via Spearman's correlation analysis. We collected patients' clinical and pathological data. Univariate and multivariate analyses were performed to analyze the factors influencing survival. Results We included 157 patients with advanced NSCLC. Among these, 135 died, 13 survived, and nine were lost to follow-up (median follow-up period, 69 months). SUVmax was correlated with lesion size and was significantly greater for tumors ≥3 cm than for tumors 6 (HR = 0.651, 95% confidence interval, 0.436-0.972; Wald value, 4.400; p = 0.036). Conclusions The SUVmax of the primary lung lesion on PET/CT is significantly correlated with survival in treatment-naive patients with advanced NSCLC.

Global warming is one of the key limiting factors affecting the cultivation of Pyropia haitanensis which is an economically important macroalgae species grown in southern China. However, the mechanism underlying the high-temperature tolerance of P. haitanensis remains largely unknown. In a previous study, we showed that the expression of the small heat shock protein 22 gene (Hsp22) is upregulated in P. haitanensis in response to high-temperature stress, but the associated regulatory mechanism was not fully elucidated. In this study, a transgenic Chlamydomonas reinhardtii expression system was used to functionally characterize P. haitanensis Hsp22. Our analyses indicated that the C-terminal of PhHsp22 is highly conserved and contains an A-crystal structure domain. A phylogenetic analysis revealed PhHsp22 is not closely related to small heat shock protein genes in other species. Additionally, PhHsp22 expression significantly increased at 3 and 6 h after initiating 33 °C treatment, which improved the survival rate of transgenic C. reinhardtii during the early stage of high-temperature treatment. The further transcriptome analysis revealed that PhHsp22 expression can promote pathways related to energy metabolism, metabolites metabolism, and protein homeostasis in transgenic C. reinhardtii cells exposed to high temperatures. Therefore, PhHsp22 may be crucial for the response of Pyropia species to high-temperature stress. Furthermore, this gene may be useful for breeding new high-temperature algal strains.

Objectives To study the feasibility of using unbalanced embryos as a reference in distinguishing euploid carrier and noncarrier embryos by single nucleotide polymorphism (SNP) array-based preimplantation genetic testing (PGT) for reciprocal translocations. Methods After comprehensive chromosome screening (CCS), euploid embryos were identified as normal or carriers using a family member as a reference. Next, unbalanced embryos were used as a reference, and the results were compared with the previous ones. Karyotypes of transferred embryos were validated by prenatal diagnosis. Results Of 995 embryos from 110 couples, 288 were found to be euploid. Using a family member as a reference, 142 and 144 embryos were tested to be euploid noncarrier and carrier respectively, and the remaining 2 embryos were undetermined. When unbalanced embryos were selected as references, all the results were consistent with the previous ones. A total of 107 embryos were transferred, resulting in 66 clinical pregnancies. Karyotypes of prenatal diagnosis were all in accordance with the results of tested embryos. Conclusions SNP array-based haplotyping is a rapid and effective way to distinguish between euploid carrier and noncarrier embryos. In case no family member is available as a reference, unbalanced embryos can be used for identification of euploid carrier and noncarrier embryos.