The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels ; Carbohydrates ; chemistry ; Cellulose ; chemistry ; Ethanol ; metabolism ; Fermentation ; Furaldehyde ; chemistry ; Glycerol ; chemistry ; Hydrolysis ; Triticum

Objective: To investigate the successful signs of laser photocoagulation or endolaser combined vitrectomy for congenital optic disc pit (ODP) with serous macular detachment. Methods: A retrospective case analysis. Twelve eyes of 12 patients with congenital ODP complicated with serous macular detachment diagnosed in Zhongshan Ophthalmic Center from 2003 to 2018 were included in this study. There were 2 males (2 eyes) and 8 females (8 eyes). The average age was 30.17 years old. Retinal laser photocoagulation and/or vitrectomy were performed in all the eyes. The optic disc and macular area were scanned using the tracking mode of the Germany Heidelberg Spectralis OCT instrument. The "dam-sign" change was a retinal choroidal scar on the channel between the ODP and the macula on the OCT image after treatment. The eyes were divided into a "dam-sign" change group (group A, 10 eyes) and no "dam-sign" change group (group B, 2 eyes). The BCVA of eyes in group A was 0.03-0.6. In group A, 1 eye was treated with laser photocoagulation alone, 9 eyes were treated with vitrectomy combined with laser photocoagulation. The BCVA of eyes in group B was 0.1. All the eyes in group B underwent vitrectomy combined with laser photocoagulation. The follow-up was ranged from 6 to 174 months. The same equipment and method before treatment were used for 1, 3, 6, 12, 18, and 24 months after treatment. The BCVA, absorption of subretinal fluid (SRF) and the formation of "dam-sign" change were observed. Results: At the last follow-up, the BCVA of eyes in group A was 0.4-1.0 (0.4 in 1 eye, 0.5 in 3 eyes, 0.6 in 2 eyes, 0.8 in 3 eyes, 1.0 in 1 eye). In 9 eyes treated with vitrectomy combined with laser photocoagulation, SRF was completely absorbed. In 1 eye treated with laser photocoagulation alone, SRF remained in small amount. The BCVA of the two eyes in group B was 0.03 and 0.3, respectively; and SRF was not absorbed in both of them. Conclusions The "dam-sign" change near optic disc after laser photocoagulation can promote SRF absorption and improve BCVA. It can be used as a success indicator after treatment.

Background: and Purpose: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe central nervous system disorder mediated by NMDAR antibodies that damages neurons. We investigated the correlation between cytoskeletal autoantibodies and the clinical severity in patients with anti-NMDAR encephalitis. Methods: Non-NMDAR autoantibodies were identified by screening matched cerebrospinal fluid (CSF) and the serum samples of 45 consecutive patients with anti-NMDAR encephalitis and 60 healthy individuals against N-methyl-D-aspartate receptor 1-transfected and nontransfected human embryonic kidney 293T cells. Immunocytochemistry was performed to assess antibody binding in rat brain sections and primary cortical neurons. Cell-based assays and Western blotting were applied to identify autoantibodies targeting medium neurofilaments (NFMs). We compared clinical characteristics between patients with NMDAR encephalitis who were positive and negative for anti-NFM-autoantibodies. Results: Anti-NFM autoantibodies were detected in both the serum and CSF in one patient (2%) and in the serum only in six patients (13%). No antibodies were detected in the serum of healthy controls (7/45 vs. 0/60, p=0.0016). Four of the seven patients with anti-NFM autoantibodies in serum were children (57%), and three (43%) had abnormalities in brain magnetic resonance imaging. These patients responded well to immunotherapy, and either no significant or only mild disability was observed at the last follow-up. Anti-NMDAR encephalitis did not differ with the presence of anti-NFM autoantibodies. Conclusions Anti-NFM autoantibodies may be present in patients with anti-NMDAR encephalitis, indicating underlying neuronal damage. A large cohort study is warranted to investigate the clinical differences between patients with NMDAR encephalitis according to their antiNFM antibody status.

Objective To investigate the efficacy and influential factors of post-resection 131 I thera-py in DTC patients with metastases can′t be removed by surgery. Methods A total of 112 DTC patients ( 41 males, 71 females, age range 8-80 years) with metastases from January 2012 to October 2016 in Affil-iated Hospital of Qingdao University were retrospectively analyzed. All patients underwent post-resection 131 I therapy, and the therapeutic effect was assessed. Univariate analysis (χ2 test, rank sum test) and the logis-tic regression analysis were performed to select the influential factors for treatment effect. The ROC curve a-nalysis of significant factors to predict the ineffectiveness of 131 I therapy was performed. Results Patients underwent single or repeated 131I therapy (1-9 times), and the effective rate was 75.89%(85/112). Univa-riate analysis showed that the tumor diameter, the location of metastasis, preablation sTg and sTg/TSH ratio were influential factors for 131 I therapy on metastases(χ2 values:7.187, 15.381;z values:-5.053,-5.187, all P<0.05) . Logistic regression analysis showed that the tumor diameter, preablation sTg were independent influential factors for 131 I therapy ( both P<0.05) . The areas under ROC curve for sTg, sTg/TSH ratio, and tumor diameter to predictive ineffectiveness were 0.824, 0.832, and 0.718, respectively. The cutoff values were 29.825μg/L, 0.298, 3.250 cm, respectively, the sensitivities were 92.60%, 96.30%, 40.70%, and the specificities were 62.40%, 55.30%, 91.80%. Conclusions The therapeutic effect of 131I therapy on metastases from DTC is significant. The longer tumor diameter, the distant metastasis, and the higher preab-lation sTg are negative influential factors for131 I therapy, which could be used to predict the therapeutic effect.

Objective To investigate the regulation of mesenchymal stem cells (MSCs) on the expression of tumor necrosis factor-α-induced protein 8-like 2 (TNFAIP8L2, TIPE2) in the peripheral blood-derived macrophages of systemic lupus erythematosus (SLE) patients. Methods The expression of TIPE2 mRNA in peripheral blood mononuclear cells (PBMCs) and macrophages from SLE patients were detected by Quantitative polymerase chain reaction (qPCR). The TIPE2 protein levels in SLE PBMCs and peripheral blood-derived macrophages were detected by western blot, flow cytometry and immunofluorescence staining, respec-tively. Data were analyzed by t test and Spearman correlation analysis. Results The TIPE2 mRNA expres-sion in PBMCs of SLE patients was significantly lower than that of healthy controls [(0.41 ±0.14) vs (1.06±0.39), t=5.376, P<0.01], as well as the TIPE2 protein level [(0.40 ±0.21) vs (1.09 ±0.26), t=2.963, P<0.05]. The expression of TIPE2 mRNA in peripheral blood-derived macrophages from SLE patients was significantly decreased [(0.56±0.24) vs (1.07±0.38), t=5.203, P<0.01). Moreover, TIPE2 mRNA level of peripheral blood-derived macrophages was negatively correlated with systemic lupus erythematosus disease activity index (SLEDAI) score (r=-0.60, P<0.01), 24-hour urinary protein (r=-0.46, P<0.05) and erythrocyte sedimentation rate (r=-0.46, P<0.05) in SLE patients. The percentage of TIPE2+cells in peripheral blood-derived macrophages (TIPE2+/CD14+)％ from SLE patients was significantly lower than that in healthy controls [(51.4 ±18.5)％ vs (82.4 ±7.5)％, t=2.679, P<0.05]. After 24 hours co-cultured with MSCs, the TIPE2 mRNA expression in SLE per-ipheral blood-derived macrophages was significantly increased [(2.2 ±0.7) vs (1.0 ±0.3), t=3.729, P<0.05). Immunofluorescence results showed the same increase of TIPE2 protein in SLE peripheral blood-derived macrophages [(0.112 ±0.020) vs (0.074 ±0.016), t=3.268, P<0.05]. Conclusion The TIPE2 level in peripheral blood-derived macrophages of SLE patients are decreased. MSCs upregulate the TIPE2 expression in vitro, suggesting that TIPE2 can be a new target for MSCs in the treatment of SLE.

Objective:To develop a single-tube one-step multiplex nested real-time reverse transcription polymerase chain reaction (RT-PCR) assay for the simultaneous detection of 2019-nCoV, influenza A virus, influenza B virus and internal-control with human-derived gene.Methods:This study included 30 positive specimens for 2019-nCoV nucleic acid detection and 336 screening specimens collected from the arrivals at Guangzhou Baiyun Airport between February 2020 and February 2022. Sixty-four positive specimens of other respiratory pathogens were also collected from the arrivals at Guangzhou Baiyun Airport during the three-year period before the occurrence of COVID19 outbreak in 2020, and 7 positive viral strains of respiratory pathogens were provided by collaborative laboratories. In order to establish a set of multiplex nested real-time RT-PCR assay, a group of primers and probe combinations for a multiplex nested real-time RT-PCR was designed and screened according to a selection of nucleotide conserved regions of the ORF and N genes of 2019-nCoV and the M gene of influenza A and B viruses, while nested amplification primers and probe for the internal-control with human-derived gene were introduced. Then the prepared pseudovirus-positive quality control and sample discs were applied to evaluate the sensitivity and specificity. Clinical specimens were performed to validate the applicability of the method.Results:The results show that the established one-step multiplex nested real-time RT-PCR assay can specifically detect 2019-nCoV and influenza A and B viruses, with the limit-of-detection of about 125 copies/ml for 2019-nCoV and about 250 copies/ml for influenza A and B viruses. Totally 101 positive samples of various respiratory pathogens were detected, showing that the detection sensitivities of 2019-nCoV and influenza A and B viruses were 96.67%, 92.86% and 96.15%, respectively, with the specificity of 100%. No false-positive detection was found in the applied detection of more than 300 clinical samples.Conclusions:A one-step multiplex nested real-time RT-PCR assay for 2019-nCoV, influenza A and B viruses and human-derived gene internal-control was developed. The assay has good sensitivity and specificity and can be used for rapid screening of 2019-nCoV and influenza A and B viruses in high-volume samples.

OBJECTIVE To establish the fing erprint of Temurin- 5 powder，conduct chemical pattern recognition analysis ，and determine the contents of 4 components simultaneously. METHODS The fingerprints of 10 batches of Temurin- 5 powder were established and similarity evaluation was performed by using high performance liquid chromatography （HPLC）combined with the Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine （2012 edition）；common peaks were identified by comparing with mixed substance control. The common peaks were analyzed by systematic cluster analysis and principal component analysis with SPSS 26.0 software. The HPLC method was used to determine the contents of gallic acid ， geniposide，chlorogenic acid and ellagic acid in 10 batches of samples. RESULTS A total of 15 common peaks were identified from the fingerprints of 10 batches of Temurin-5 powder，and the similarity was 0.997-0.999. It was identified that peak 1 was gallic acid ，peak 3 was geniposide ，peak 5 was chlorogenic acid and peak 12 was ellagic acid. Among the 10 batches of samples ， S4 and S 9 were grouped into one category ，S6-S8 were grouped into one category ，and the other batches of samples were grouped into one category. The accumulative variance contribution rate of first three principal components was 89.245％. The linear ranges of gallic acid ，geniposide，chlorogenic acid and ellagic acid were 5.55-177.5，15.98-511.5，2.56-82.0 and 13.48-431.5 μg/mL， respectively. RSDs of precision ，stability（24 h）and repeatability tests were all less than 2％（n＝6 or n＝7）. The average recoveries were 101.56％，102.21％，98.60％ and 96.62％，respectively，RSDs were 1.90％，1.61％，1.58％ and 1.73％（n＝6）. Average contents of above components were 5.03-5.64，10.38-12.16，1.40-1.69，6.47-7.11 mg/g，respectively. CONCLUSIONS The established fingerprint is stable and feasible ，and the content determination method meets the relevant regulations. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Temurin- 5 powder.

OBJECTIVE To explore the intestinal absorption characteristics of saikosaponins. METHODS Based on everted intestinal sac model， using accumulative absorption amount （Q） and absorption rate constant （Ka） as indexes， UHPLC-MS/MS technique as a method， the absorption of saikosaponin A， B2， C， D and F from total saponins of Bupleurum chinense （8 g/mL， by crude drug） in the duodenum， jejunum and ileum was detected. RESULTS The correlation coefficients （r） of the regression equations for the absorption of saikosaponins A， B2， C and F in the duodenum， jejunum and ileum were all higher than 0.95， while the r of saikosaponin D in the above intestinal segments was lower than 0.95； compared with the absorption of the same composition in the duodenum， the Q and Ka of saikosaponin A and C circulating in jejunum and ileum for 120 min， as well as the Q and Ka of saikosaponin F circulating in the ileum for 120 min were significantly decreased （P＜0.05）. CONCLUSIONS Saikosaponin A and the other 4 saikosaponins are all absorbed in the duodenum， jejunum and ileum； among them， saikosaponin A， B2， C and F are linearly absorbed， which conforms to the zero-order absorption characteristics， but saikosaponin D shows non- linear absorption.

Objective : To explore the association of time in range(TIR) and glucose management indicator ( GMI) with the risk of type 2 diabetic nephropathy (DN) ． Methods : The clinical data of 215 patients with type 2 diabetes mellitus (T2DM) were collected and analyzed．According to the results of estimated glomerular filtration rate (eGFR) and urinary albumin to creatinine ratio( UACR) ，they were divided into 117 patients with T2DM and 98 patients with DN．The clinical data，biochemical indicators and continuous glucose monitoring ( CGM) indicators of the two groups were compared．Logistic regression was used to analyze the influencing factors of DN risk．The predictive value of TIR and GMI on the risk of DN was evaluated by receiver operating characteristic (ROC) curve. Results: There were significant differences in age，duration of diabetes，systolic blood pressure，glycosylated hemoglobin ( HbA1c) ，fasting plasma glucose (FPG) ，2 hour postprandial plasma glucose (2hPG) ，creatinine( Cr) ，UACR, eGFR between the two groups(P＜0. 05) ．There were statistically significant differences between the two groups in the CGM indexes of GMI，mean absolute difference of mean of daily differences ( MODD) ，glucose above target range time(TAR) and TIR(P＜0. 05) ．The results of logistic regression analysis showed that TIR was a protective factor of DN．In the ROC curve analysis of TIR prediction DN，the area under the ROC curve was 0. 718 (95% CI = 0. 648 ～0. 789，P＜0. 001) ，and the Yoden index was 0. 38．At this time，the sensitivity was 66. 7% ，and the specificity was 71. 3%．In the ROC curve analysis of GMI prediction DN，the area under the ROC curve was 0. 701 (95% CI = 0. 629 ～0. 774，P＜0. 001) ，and the Yoden index was 0. 368．At this time，the sensitivity was 63. 3% ， and the specificity was 73. 5%． Conclusion Specifically，lower TIR and higher GMI increase the risk of DN.

Increasing evidences suggest the important role of calcium homeostasis in hallmarks of cancer, but its function and regulatory network in metastasis remain unclear. A comprehensive investigation of key regulators in cancer metastasis is urgently needed. Transcriptome sequencing (RNA-seq) of primary esophageal squamous cell carcinoma (ESCC) and matched metastatic tissues and a series of gain/loss-of-function experiments identified potassium channel tetramerization domain containing 4 (KCTD4) as a driver of cancer metastasis. KCTD4 expression was found upregulated in metastatic ESCC. High KCTD4 expression is associated with poor prognosis in patients with ESCC and contributes to cancer metastasis in vitro and in vivo. Mechanistically, KCTD4 binds to CLIC1 and disrupts its dimerization, thus increasing intracellular Ca²⁺ level to enhance NFATc1-dependent fibronectin transcription. KCTD4-induced fibronectin secretion activates fibroblasts in a paracrine manner, which in turn promotes cancer cell invasion via MMP24 signaling as positive feedback. Furthermore, a lead compound K279-0738 significantly suppresses cancer metastasis by targeting the KCTD4‒CLIC1 interaction, providing a potential therapeutic strategy. Taken together, our study not only uncovers KCTD4 as a regulator of calcium homeostasis, but also reveals KCTD4/CLIC1-Ca²⁺-NFATc1-fibronectin signaling as a novel mechanism of cancer metastasis. These findings validate KCTD4 as a potential prognostic biomarker and therapeutic target for ESCC.