ObjectiveTo observe the clinical efficacy of Sanren Runchang formula in treating constipation-predominant irritable bowel syndrome （IBS-C） by regulating the brain-gut axis and the effects of the formula on serum levels of 5-hydroxytryptamine （5-HT）， vasoactive intestinal peptide （VIP）， and substance P （SP）. MethodsA randomized controlled design was adopted， and 72 IBS-C patients meeting Rome Ⅳ criteria were randomized into observation and control groups （36 cases）.The observation group received Sanren Runchang formula granules twice daily， and the control group received lactulose oral solution daily for 4 weeks. IBS Symptom Severity Scale （IBS-SSS）， IBS Quality of Life Scale （IBS-QOL）， and Bristol Stool Form Scale （BSFS） were used to assess clinical symptoms， and bowel movement frequency was recorded. The Self-Rating Anxiety Scale （SAS） and Self-Rating Depression Scale （SDS） were employed to evaluate psychological status. ELISA was employed to measure the serum levels of 5-HT， VIP， and SP. ResultsThe total response rate in the observation group was 91.67% （33/36）， which was higher than that （77.78%， 28/36） in the control group （χ²=4.50， P<0.05）. After treatment， both groups showed increased defecation frequency and BSFS scores， decreased IBS-SSS total score， abdominal pain and bloating scores， IBS-QOL health anxiety， anxiety， food avoidance， and behavioral disorders scores， SAS and SDS scores， serum 5-HT and VIP levels， and increased SP levels （P<0.05， P<0.01）. Moreover， the observation group showed more significant changes in the indicators above than the control group （P<0.05， P<0.01）. The SP level showed no significant difference between the two groups. During the 4-week follow-up， the recurrence rate was 5.88% in the observation group and 31.25% in the control group. No adverse events occurred in observation group， and 2 cases of mild diarrhea occurred in the control group. ConclusionSanren Runchang formula demonstrated definitive efficacy in alleviating gastrointestinal symptoms and improving the psychological status and quality of life in IBS-C patients， with a low recurrence rate. The formula can regulate serum levels of neurotransmitters such as 5-HT and VIP， suggesting its potential regulatory effect on the brain-gut axis through modulating neurotransmitters and neuropeptides. However， its complete mechanism of action requires further investigation through detection of additional brain-gut axis-related biomarkers.

Cerebral small vessel disease （CSVD） is an important etiology of vascular cognitive impairment and stroke， and there are still many challenges in its early diagnosis and pathological mechanisms. As a type of extracellular vesicles that exist stably in body fluids and can cross the blood-brain barrier， exosomes have attracted wide attention in recent years. Studies have shown that various exosomal proteins have a potential value in the diagnosis and treatment of CSVD and related cognitive impairment， providing new research directions for the noninvasive management of these diseases. This article systematically reviews the latest advances in exosomal proteins for the diagnosis and treatment of CSVD and related cognitive impairment， in order to provide new perspectives for the application of exosomal proteins in the precise treatment of CSVD.
Biomarkers

Objective:To examine the effects of Zishenwan Prescription on bile acid metabolism in mice with diabetic encephalopathy; To explore its mechanism of improvement of diabetic encephalopathy.Methods:Male C57BL/6J mice were used to replicate the mouse model of type 2 diabetes mellitus by using high-fat chow and a single intraperitoneal injection of streptozotocin (120 mg/kg). The mice were screened for diabetic encephalopathy by using the Morris water maze test after 8 weeks of continuous stimulation with hyperglycemia, and were divided into model group and Zishenwan Prescription group according to random number table method, with 12 mice in each group. The mice in the Zishenwan Prescription group were treated with the crude extract of Zishenwan Prescription (9.36 g/kg) by gavage, and the normal group and the model group were treated with the same volume of distilled water once a day for 8 weeks. At the end of the treatment, Morris water maze test was used to investigate the cognitive function of diabetic encephalopathy mice; cresyl violet staining was used to detect the number of granule neurons in the hippocampus; serum and feces were collected to detect the content of bile acids by liquid-liquid coupling; hepatic bile acid synthase CYP7a1 and CYP27a1, farnesol X receptor (FXR), fibroblast growth factor 15 (FGF15), fibroblast growth factor receptor 4 (FGFR4), and ileocecal apical sodium-dependent bile acid transporter protein (ABST) mRNA levels were detected by using fluorescence quantitative PCR assay.Results:Compared with the model group, mice in the Zishenwan Prescription group had shorter evasion latency time ( P<0.05 or P<0.01), decreased time to first reach the platform ( P<0.01), increased number of times to traverse the platform ( P<0.01), and reduced neuronal cell damage in hippocampal area; mice in the Zishenwan Prescription group showed decreased serum and fecal total bile acid content ( P<0.05 or P<0.01); the liver CYP7a1 and CYP27a1 mRNA expressions increased ( P<0.01), and FXR and FGF15 mRNA expressions decreased ( P<0.01); ileal ABST mRNA expression decreased ( P<0.01). Conclusion:Zishenwan Prescription may regulate bile acid metabolism, inhibit FRX-FGF15/FGFR4 signaling and ABST expression to promote new bile acid synthesis and conjugated bile acid reabsorption, and thus improve cognitive function in diabetic encephalopathy mice.

L-asparaginase (L-ASN) is widely applied in the treatment of malignant tumor and low-acrylamide food production, however, the low expression level hampers its application. Heterologous expression is an effective strategy to increase the expression level of target enzymes, and Bacillus is generally used as the host for efficient production of enzymes. In this study, the expression level of L-asparaginase in Bacillus was enhanced through optimization of expression element and host. Firstly, five signal peptides (SP_SacC, SP_AmyL, SP_AprE, SP_YwbN and SP_WapA) were screened, among which SP_SacC showed the best performance, reaching an activity of 157.61 U/mL. Subsequently, four strong promoters (P43, P_ykzA-P43, P_Ubay and P_bacA) from Bacillus were screened, and tandem promoter P_ykzA-P43 showed the highest yield of L-asparaginase, which was 52.94% higher than that of control strain. Finally, three Bacillus expression hosts (B. licheniformis Δ0F3 and BL10, B. subtilis WB800) were investigated, and the maximum L-asparaginase activity, 438.3 U/mL, was reached by B. licheniformis BL10, which was an 81.83% increase compared with that of the control. This is also the highest level of L-asparaginase in shake flask reported to date. Taken together, this study constructed a B. licheniformis strain BL10/P_ykzA-P43-SP_SacC-ansZ capable of efficiently producing L-asparaginase, which laid the foundation for industrial production of L-asparaginase.
Bacillus licheniformis/metabolism* ; Asparaginase/genetics* ; Bacillus/genetics* ; Protein Sorting Signals ; Promoter Regions, Genetic/genetics* ; Bacillus subtilis/genetics* ; Bacterial Proteins

Numerous studies on cancers, biopharmaceuticals, and clinical trials have necessitated comprehensive and precise analysis of protein O-glycosylation. However, the lack of updated and convenient databases deters the storage of and reference to emerging O-glycoprotein data. To resolve this issue, an O-glycoprotein repository named OGP was established in this work. It was constructed with a collection of O-glycoprotein data from different sources. OGP contains 9354 O-glycosylation sites and 11,633 site-specific O-glycans mapping to 2133 O-glycoproteins, and it is the largest O-glycoprotein repository thus far. Based on the recorded O-glycosylation sites, an O-glycosylation site prediction tool was developed. Moreover, an OGP-based website is already available (http://www.oglyp.org/). The website comprises four specially designed and user-friendly modules:statistical analysis, database search, site prediction, and data submission. The first version of OGP repository and the website allow users to obtain various O-glycoprotein-related information, such as protein accession Nos., O-glycosylation sites, O-glycopeptide sequences, site-specific O-glycan structures, experimental methods, and potential O-glycosylation sites. O-glycosylation data mining can be performed efficiently on this website, which will greatly facilitate related studies. In addition, the database is accessible from OGP website (http://www.oglyp.org/download.php).

Currently, the value of adjuvant therapy after radical resection of esophageal squamous cell carcinoma remains elusive. Some studies have reported that radiotherapy can improve the locoregional control rate and overall survival of patients. However, the design of radiotherapy target area, intervention time and dose of radiotherapy are controversial. In this article, literature review was conducted and the current status and controversy of adjuvant radiotherapy after radical resection of esophageal squamous cell carcinoma were reviewed.

Objective To detect the anti-citrullinated alpha-enolase peptide 1 (CEP-1) antibody in rheumatoid arthritis (RA). Methods One hundred and twenty-nine patients with RA were enrolled randomly. Thirty-one patients with primary Sj?gren's syndrome (pSS), 32 patients with systemic lupus erythematosus (SLE), 32 patients with osteoarthritis (OA), and 106 healthy controls (HC) were include into this study. Anti-CEP-1 antibody was detected by enzyme-linked immunosorbent assay (ELISA). The correlations between serum anti-CEP-1 antibody and clinical features, disease activities,laboratory tests or Sharp scores of RA patients were evaluated. Mann-Whitney U test and χ2 test were used for statistical analysis. Results ①Anti-CEP-1 antibodies were positive in 64.3%(83/129) of RA patients, 22.6%(7/32) of pSS patients, 12.5%(4/32) of SLE patients, none of OA patients (0/32) or healthy controls. The positivity of anti-CEP-1 antibody was significantly higher than those in pSS ( χ2=17.7), SLE ( χ2=25.7), OA ( χ2=42.5), and healthy controls ( χ2=102.6) (P<0.01, respectively). The specificity of anti-CEP-1 antibody in RA was 94.5%. ②In patients without anti-citrullinated protein/peptide autoantibodies (ACPA), rheumatoid factor (RF) or the patients without ACPA and RF, the positive rate of anti-CEP-1 antibody was 30.3%(10/33), 41.9%(18/43) and 22.7%(5/22), respectively. ③Compared with patients without anti-CEP-1 antibodies, patients with anti-CEP-1 anti-bodies had higher rates of joint deformity, bone erosion and high disease activities (P<0.05, respectively). ④ Higher rate of interstitial lung disease (ILD) was found in RA patients with anti-CEP-1 antibody (19.3% vs 4.3%, χ2=5.494, P<0.05). ⑤The patients with anti-CEP-1 anti-body had higher rates of elevated erythrocyte sedimentation rate (ESR) ( χ2=6.543) and decreased serum albumin ( χ2=6.59), compared to patients without anti-CEP-1 antibody (P<0.05, respectively). Conclusion Anti-CEP-1 antibody has high sensitivity and specificity for RA diagnosis. Combination of anti-CEP-1 antibody with other RA antibodies might improve the early diagnosis of RA. Anti-CEP-1 antibody is significantly associated with joint damage, disease activity and pulmonary interstitial fibrosis.

Objective To compare the relationship of acceptable noise level (ANL) between monaural and binaural hearing aid in patients with bilateral moderate-to-severe hearing loss, and to investigate the clinical significance of the ANL in binaural hearing aid fitting and the predictive role in the hearing aid effect assessment.Methods A total of 15 patients with bilateral moderate-to-severe hearing loss were selected, and the most comfortable levels (MCL), background noise level (BNL) and calculate ANL were tested, respectively, in 4 conditions: without hearing aids, fitted only left ears, fitted only right ears and binaural fitting.Results The ANL in 15 subjects measured at 4 conditions were 18.87±5.26, 12.60±2.47, 12.00±2.90, and 5.13±1.25 dB S/N, respectively.The MCLs were 80.40±9.28, 63.73±5.15, 62.27±5.36, and 61.80±6.05 dB HL, respectively.The BNLs were 61.67±6.14, 51.13±3.94, 50.27±4.50, and 56.67±5.16 dB HL.The ANL difference between the only left and right fitting groups was not statistically significant(P>0.05).The ANL difference between the monaural or the binaural hearing aid group and without hearing aids group were statistically significant (P<0.05), respectively.Compared with the monaural hearing aid group, the binaural hearing aid group had significantly lower ANL(P<0.05).Conclusion For people with bilateral hearing loss, hearing aids can improve their ability to manage the background noise, and binaural hearing aid fitting is better than monaural.

Objective To investigate any potential and independent demographic and serologic risk factors contributing to bone destruction in patients with rheumatoid arthritis ( RA) . Methods A total of 445 patients with RA were recruited in this study. Three autoantibodies including rheumatoid factor ( RF) , anti-cyclic citrullinated peptide antibody ( anti-CCP antibody) and anti-citrullinated alpha-enolase peptide 1 antibody ( anti-CEP-1 antibody) were quantified by using specific ELISA kits. The hand radiographs of all subjects were graded by using the modified Sharp/van der Heijde score ( Sharp score) . The potential and in-dependent risk factors were assessed by using univariate linear regression analyses and the stepwise multiple regression analysis, respectively. Results Based upon the univariate regression analyses, 7 covariates were identified as the potential risk factors for bone destruction in patients with RA, which were female (β=0. 100, P=0. 035), longer disease duration (β=0. 498, P=3. 26×10-29), RF (β=0. 096, P=0. 042), younger age at onset (β=-0. 312, P=1. 60 × 10-11 ), anti-CCP antibody positive (β=0. 202, P=1.74×10-5), anti-CEP-1 antibody positive (β=0.148, P=0.017) and positive for either anti-CCP or anti-CEP-1 antibodies (β=0. 157, P=1. 42×10-3). However, smoking (β=-0. 121, P=0. 018) were identi-fied as the potential protective factors. The multiple regression analysis indicated that the longer disease du-ration (P=2. 24×10-15) and anti-CCP antibody positive (P=0. 012) were independent risk factors for bone destruction. Conclusion Female, longer disease duration, younger age at onset, RF, anti-CCP and anti-CEP-1antibodies are potential risk factors for bone damage in patients with RA. Moreover, longer disease du-ration and anti-CCP antibody are two independent risk factors contributing to bone destruction in RA.

Osteosarcoma ( OS) is the most common primary malignant bone tumor in children and adolescents. Currently, the combination of surgical resection and chemotherapy is the main method of treatment for OS in the clinic.Al-though, the survival rate has been greatly improved in OS patients with localized disease, the 5-year survival rate has re-mained <20%.It is very important to understand the cause of disease, to develop novel drugs or therapeutics, and to learn the disease through animal models.This review will introduce the progress of animal models of osteosarcoma over the last years.