Human keratinocyte growth factor-1 (hKGF-1), a member of the fibroblast growth factor (FGF) family, plays crucial roles in organ development, cell proliferation, wound healing, and tissue repair, representing one of the most effective and specific growth factors for skin repair. However, obtaining recombinant hKGF-1 remains challenging due to its universally low expression efficiency in vitro. This study employs the Bombyx mori baculovirus expression system to establish a technological platform that utilizes the economically important insect Bombyx mori as a bioreactor for high-efficiency and low-cost expression and production of recombinant human keratinocyte growth factor 1 (hKGF-1) protein, ultimately achieving high-level expression of hKGF-1 in Bombyx mori ovary cell line (BmN). In this study, we optimized the hKGF-1 sequence based on the codon preference of baculovirus. By fusing hKGF-1 with polyhedrin (highly expressed in this system) and adding extra promoters and enhancers, we significantly improved the expreesion level of hKGF-1 in Bombyx mori cells. The results demonstrated that the aforementioned strategies significantly enhanced the expression level of hKGF-1 in Bombyx mori cells. SDS-PAGE and Western blotting results revealed that the highest hKGF-1 expression (accounting for 8.7% of total cellular protein) was achieved when the Polh promoter was combined in tandem with the P6.9 promoter and hKGF-1 was fused with a 15-residue polyhedrin fragment for co-expression. The optimal harvest time was determined to be 120 h post transfection. This study achieved the efficient expression of hKGF-1 in Bombyx mori cells, establishing an ideal technological platform for the industrial utilization of recombinant hKGF-1. The developed methodology not only provides valuable technical references for the production of other growth factors and complex proteins, but also demonstrates significant implications for employing silkworms as bioreactors for recombinant human protein expression.
Bombyx/metabolism* ; Animals ; Baculoviridae/metabolism* ; Humans ; Fibroblast Growth Factor 7/biosynthesis* ; Recombinant Proteins/genetics* ; Cell Line ; Genetic Vectors/genetics*

Bombyx mori nucleopolyhedrovirus (BmNPV) is extremely harmful to the silk industry. The polyhedrin, which encodes the polyhedrin (Polh), can be expressed at ultra-high levels and form occlusion bodies in the nucleus, embedding the progeny virus within it. However, the detailed mechanism by which polyhedrin is transported into the host cell nucleus remains unknown. Clarifying the nuclear import mechanisms of viral proteins can help us develop better prevention and treatment measures against baculoviruses. This study employed molecular cloning, co-immunoprecipitation, and immunofluorescence to analyze in detail the expression pattern of the highly expressed polyhedrin in the very late stage of the virus, and further revealed that the host protein importin α participates in the nuclear import of polyhedrin through protein interactions. This study provides a reference for further elucidating the nuclear import mechanisms of the baculovirus proteins including polyhedrin that can enter the nucleus.
Nucleopolyhedroviruses/metabolism* ; Active Transport, Cell Nucleus ; Animals ; Bombyx/virology* ; alpha Karyopherins/metabolism* ; Cell Nucleus/metabolism* ; Viral Structural Proteins/metabolism* ; Occlusion Body Matrix Proteins

OBJECTIVE To study the protective effect and possible mechanism of soybean isoflavones against threatened miscarriage rats. METHODS Female mice were selected to promote estrus and mate with male mice. After pregnancy， they were randomly divided into normal group （purified water， i.g.， n＝10）， model group （purified water， i.g.， n＝9）， positive control drug group （progesterone 4 mg/kg， i.m.， n＝9）， low-， medium- and high-dose soybean isoflavone groups （25， 50 and 100 mg/kg， i.g.， n＝10）. Except for the normal group， the rest were given mifepristone+misoprostol on the 8th day of pregnancy to establish threatened miscarriage model， and then given purified water or drugs， once a day， on days 1-7 and 9-12 of pregnancy， respectively. At 14 days of pregnancy， the rates of fetal protection were counted. Serum levels of β-human chorionic gonadotrophin （β-HCG） and progesterone （P） in rats were detected. Pathological and morphological changes in rat placenta and decidua tissues were observed， and the apoptosis indexes of cells were detected； mRNA and protein expressions of factor of apoptosis related （Fas）， factor of apoptosis related ligand （FasL）， proliferating cell nuclear antigen （PCNA） and heparin binding epidermal growth factor （HB-EGF） were determined in placenta tissues， and mRNA and protein expressions of Fas， PCNA and HB-EGF in decidua tissues were detected. RESULTS In the model group， the placental tissues of rats were hyperemia and dilatation， with fewer and irregular blood vessels； severe stromal edema，inflammatory cell infiltration and iron-choledrin depositionwere observed. Compared with model group， the fetal survival rates， serum levels of β-HCG and P， the expressions of PCNA and HB-EGF mRNA and proteins in the placenta and decidua tissue of soybean isoflavone groups increased significantly （P＜ 0.05）， while pathological changes were improved significantly； cell apoptosis index in the placenta and decidua tissue， the expressions of Fas， FasL mRNA and proteins in the placenta and Fas mRNA and protein in the decidua tissue decreased significantly （P＜0.05）. The effect of soybean isoflavones was dose-dependent （P＜0.05）. CONCLUSIONS Soybean isoflavone has protective effect on threatened miscarriage， the mechanism of which is related to down-regulating the expressions of Fas and FasL mRNA and protein at the maternal-fetal interface， and up-regulating the expressions of mRNA and protein of PCNA and HB- EGF.

Objective:To investigate the perioperative impact of closure of mesenteric fissure in laparoscopic right hemicolectomy.Methods:The prospective randomized controlled trial was conducted. The clinical data of 320 patients who underwent laparoscopic right hemicolectomy in 11 medical centers, including The First Affiliated Hospital of China Medical University et al, from November 2022 to August 2023 were selected. Based on block randomization, patients were alloca-ted into the mesenteric fissure non-closure group and the mesenteric fissure closure group. Observa-tion indicators: (1) grouping of the enrolled patients; (2) intraoperative conditions; (3) postopera-tive conditions. Measurement data with skewed distribution were represented as M( Q1, Q3) and com-parison between groups was conducted using the Mann-Whitney U test. Count data were represen-ted as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher's exact probability. Comparison of ordinal data was conducted using the rank sum test. Comparison of visual analog scores was analyzed using generalized estimating equations. Results:(1) Grouping of the enrolled patients. A total of 320 patients with colon cancer were screened for eligibility, including 156 males and 164 females, aged 68(59,73)years. All the 320 patients were allocated into the mesenteric fissure non-closure group with 164 cases and the mesenteric fissure closure group with 156 cases. There was no significant difference in the age, body mass index, American Society of Anesthesiologist score, maximum tumor diameter, anastomosis location, anastomosis method, surgical approach, range of lymph node dissection, tumor staging between the two groups ( P>0.05) and there was a significant difference in the sex between them ( P<0.05). (2) Intraoperative conditions. There was no significant difference between the mesenteric fissure closure group and the mesenteric fissure non-closure group in the volume of intraoperative blood loss, operation time, conversion to laparotomy, intraoperative complication ( P>0.05). Three patients in the mesenteric fissure non-closure group were converted to laparotomy. One patient in the mesenteric fissure closure group was converted to laparotomy, and 2 cases with intraoperative complication were mesenteric hematoma. (3) Postoperative conditions. There was no significant difference between the mesenteric fissure non-closure group and the mesenteric fissure closure group in the overall postoperative complications ( χ2=0.28, P>0.05). There was no significant difference in the occurrence of postoperative intestinal obstruction, abdominal distension, ascites, pleural effusion, gastric paralysis, anastomotic bleeding, anastomotic leakage, or surgical wound infection between the two groups ( P>0.05). There was no significant difference between the two groups in the reoperation, postoperative gastric tube replacement. There was no significant differ-ence in time to postoperative first flatus, time to postoperative initial liquid food intake, time to post-operative resumption of bowel movements, duration of postoperative hospital stay, total hospital expenses between the two groups ( Z=-0.01, 0.43, 1.04, -0.54, -0.36, P>0.05). One patient in the mesenteric fissure non-closure group received reoperation. No perioperative internal hernia or death occurred in either group. The visual analog score decreased with time in both groups. There was no significant difference in the visual analog score between the mesenteric fissure closure group and the mesenteric fissure non-closure group [ β=-0.20(-0.53,0.13), P>0.05]. Conclusion:Compared with closure of mesenteric fissure, non-closure of mesenteric fissure during laparoscopic right hemi-colectomy dose not increase perioperative complications or postoperative management risk.

Human epidermal growth factor (hEGF) can be applied in the treatment of surgical trauma (burns, scalds), tissue repair, skin moisturizing, beauty, skincare, etc. However, the low expression and high cost limit the application of hEGF. In order to improve the expression level of hEGF and reduce the production cost, considering the high expression of polyhedrin, this study fused a partial sequence of polyhedrin with hEGF and expressed the fused sequence by using a silkworm baculovirus expression vector system. In view of the small molecular weight of hEGF, we connected hEGF genes in series and optimized the codons to construct multiple fusion expression vectors by fusing different partial sequences of polyhedrin at the N-terminus. The results showed that through the above strategy, the protein expression level of hEGF was significantly increased. The expression vector containing three concatenated hEGF genes with optimized codons and fused with the sequence encoding 25 or 35 residues at the N-terminus of polyhedrin showed the highest expression level.
Humans ; Epidermal Growth Factor/biosynthesis* ; Genetic Vectors/genetics* ; Recombinant Fusion Proteins/biosynthesis* ; Animals ; Bombyx/metabolism* ; Occlusion Body Matrix Proteins/genetics* ; Nucleopolyhedroviruses/genetics* ; Amino Acid Sequence

Objective:To investigate the clinical efficacy and possible mechanism of bifidobacterium tetralogy combined with escitalopram oxalate in the treatment of patients with depression and to provide evidence for further clinical research.Methods:A total of 100 patients with depression who received treatment in Taizhou Second People's Hospital from September 2019 to March 2021 were included in this study. They were randomly assigned to receive either escitalopram oxalate and placebo (control group, n = 50) or escitalopram oxalate and bifidobacterium tetralogy (observation group, n = 50). All patients received 9 weeks of treatment. Psychological status pre- and post-treatment was evaluated using the Hamilton Rating Scale for Depression and the Hamilton Rating Scale for Anxiety. Serum cortisol, inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) levels were detected using the enzyme-linked immunosorbent assay. Results:Physiological status scores in each group were significantly lower after treatment compared with before treatment. Scores of the Hamilton Rating Scale for Depression and the Hamilton Rating Scale for Anxiety scores post-treatment in the observation group were (10.78 ± 2.03) points and (6.37 ± 2.58) points, which were significantly lower than those in the control group [(16.78 ± 2.85) points, (13.23 ± 2.95) points, t = 11.40, 13.38, both P < 0.001]. Serum levels of cortisol, inflammatory factors IL-1β and TNF-α in each group were significantly decreased after treatment compared with before treatment. Serum levels of cortisol, inflammatory factors IL-1β and TNF-α post-treatment in the observation group were (137.34 ± 63.29) μg/L, (14.38 ± 6.08)ng/L, (13.95 ± 6.46) ng/L, which were significantly lower than those in the control group [(181.22 ± 59.27) μg/L, (25.94 ± 6.92) ng/L, (20.44 ± 6.24) ng/L, t = 15.29, 6.16, 9.24, all P < 0.001). Conclusion:Bifidobacterium tetralogy combined with escitalopram oxalate is highly effective on depression. The combined therapy can remarkably reduce depression and anxiety symptoms and greatly decrease serum cortisol, IL-1β, and TNF-α levels.

Objective:To study the real-world outcome of China FDA-approved Sofosbuvir (SOF)/Velpatasvir (VEL) in Northwest China.Methods:In this multicenter, prospective, real-world cohort study, we recruited patients from 10 sites from Northwest China, who were chronically infected with HCV GTs 1-6 from 06/2018 to 09/2019. Patients received SOF (400mg)/VEL (100mg) for 12 weeks, and with ribavirin 900-1200 mg for GT3 cirrhosis and for any genotype decompensated cirrhosis. The primary endpoint was sustained virological response at 12-weeks post-treatment (SVR12) and safety. The secondary endpoint was the change of liver function after the achievement of SVR12.Results:Totally, 143 patients were enrolled in the study, four patients were lost to follow-up and one died during the follow-up, 138 patients were included in per-protocol analysis. Of the 138 patients, the mean age 53 years, 53.6% male, 94.2% Han nationality, 53.6% liver cirrhosis, 10.1% HBsAg +, 6.5% renal dysfunction, 5.1% treatment-experienced, and 16.7% patients received ribavirin treatment. The genotype distribution was as follows: 35.5% GT1, 42.8% GT2, 15.9% GT3, and 5.8% un-typed. The SVR12 rate was 96.5% (138/143, 95% CI: 93.5%-99.6%) for intention-to-treat analysis, and in per-protocol analysis, all 138 patients obtained SVR12 (100%). Compared with baseline, the serum total bilirubin, ALT and AFP levels decreased (all P < 0.05), as well as increased ALB and platelet count (all P < 0.001) at post-treatment 12-weeks. Overall adverse events (AEs) rate is 29.0%, and the most common AEs were anemia (14.5%) and fatigue (8.0%). Severe side effects (edema and fatigue) occurred in 2 patients, one of whom needed a short-term interruption of treatment due to fatigue. Conclusion:In this real-world cohort study, 12-week SOF/VEL regimen with or without ribavirin achieved high SVR12 rates (96.5%-100% overall) with excellent safety profile among patients with HCV GT1/2/3 infection including patients with GT3 and cirrhosis, and led to improvement of liver function.

Objective:To explore the predictive value of regional leptomeningeal collateral circulation scale (rLMC) based on multimodal CT in recanalization of blood vessels in patients with acute ischemic stroke after thrombectomy.Methods:A retrospective analysis was conducted on clinical data of patients with acute ischemic stroke within 6 h of first onset, admitted to our hospital from October 2017 to December 2019. Before operation, the conditions of their vessels were evaluated by rLMC based on multimodal CT. Two areas, anterior cerebral artery (ACA)-middle cerebral artery (MCA) area and posterior cerebral artery (PCA)-MCA area, were divided. The total rLMC scores of two areas (0-10) were calculated: scores of 0-3, scores of 4-7, scores of 8-10. After admission, the recanalization of the blood vessels after thrombectomy was evaluated immediately according to grading of thrombolysis in cerebral infarction (TICI) after completion of thrombectomy within the time window; TICI grading≥II was defined as succeed recanalization. The correlation between rLMC scores and vascular recanalization in patients with acute ischemic stroke was evaluated.Results:Among the 80 patients, 17 were in the rLMC scores of 0-3 group, 25 in the group of rLMC scores of 4-7, and 38 in the group of rLMC scores of 8-10; 68 patients (85.00%) had vascular recanalization, and the success rate of vascular recanalization in patients from the group of rLMC scores of 8-10 was significantly higher than that in the group of rLMC scores of 0-3 (97.36% vs. 58.82%, P<0.05). Correlation analysis results showed that the rLMC score was positively correlated with success rate of vascular recanalization ( r s=0.625, P=0.000); whose sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 89.5%, 90.9%, 94.45%, 83.3% and 90.0%, respectively. Conclusion:The rLMC is closely related to the recanalization rate in patients with acute ischemic stroke after thrombectomy; the success rate of recanalization after intravascular treatment is relatively high in patients with rLMC scores of 8-10.

Objective To investigate the effect of psychological intervention combined with venlafaxine and risperidone in the treatment of refractory depression and its influence on the quality of life .Methods 97 patients with refractory depression were selected .According to the random number table , they were divided into the observation group(50 cases) and the control group(47 cases).The control group was given venlafaxine combined with risperidone treatment,the observation group received psychological intervention on basis of the control group .The Hamilton depression scale ( HAMD) of patients in the two groups before treatment ,4,8 and 12 weeks after treatment were recorded .The curative effect after 12 weeks of treatment was evaluated ,the adverse reaction was evaluated by the side effects scale ( TESS ) , and the quality of life ( WHOQOL-100 ) of the two groups before treatment , 12 weeks after treatment were compared .Results The HAMD score of the two groups had no statistically significant difference before treatment(t=0.307,P>0.05).After treatment for 4,8,12 weeks,the HAMD scores of the two groups were significantly lower than before treatment (t=5.743,17.756,27.288,4.595,15.801,23.247,all P<0.05),and the HAMD scores in the observation group after treatment for 8 weeks and 12 weeks were significantly lower than those in the control group(t=0.887,3.717,5.854,all P<0.05).After 12 weeks of treatment,the total effective rate of the observation group was significantly higher than that of the control group (90.00％ vs.74.47％,χ2 =4.043,P <0.05 ).There was no statistically significant difference in incidence rate of adverse reactions between the two groups (26.00％vs.25.52％,χ2 =0.003,P >0.05).The WHOQOL-100 scores of the two groups had no statistically significant differences before treatment(t=0.597,0.497,0.378,0.923,0.136,all P>0.05).The WHOQOL-100 scores of the two groups after treatment were significantly higher than those before treatment ( t =11.668,8.701, 6.857,7.053,2.247,4.815,4.639,2.999,4.117,2.011,all P<0.05).After treatment,the psychological and physiological independence and social relationship domain scores in the observation group were significantly higher than those in the control group(t=5.618,3.295,4.335,3.603,all P<0.05).Conclusion Psychological interven-tion combined with venlafaxine and risperidone in the treatment of refractory depression can significantly improve the clinical efficacy ,improve the clinical symptoms and the quality of life of patients .

The present study aimed to research pathogenesis and therapeutics of hepatocellular carcinoma (HCC) with the method of tissue metabolomics.A combined gas chromatography-mass spectrometry (GC-MS) method was developed suitable for analyzing the endogenous small molecule of liver cancer tissues and adjacent tissues.The unidimensional and multidimensional statistics were used to look for differential metabolites.And then,the KEGG and HMDB database were utilized to find related differential pathways and pathogenesis of HCC.The PCA and PLS-DA showed that there were significant differences on the endogenous small molecule of liver cancer tissues and adjacent tissues.Through the OPLS-Loading plot analysis,there were 25 differential metabolites and 36 relevant pathways.The differential pathways belong to carbohydrate metabolism,amino acid metabolism and mitochondrial transfer.There were 16 metabolites' area under the ROC curve which was bigger than 0.8,which were related with ATP-binding cassette (ABC) transporters,galactose metabolism,amino sugar and nucleotide sugar metabolism.It was concluded that the Warburg effect exists in HCC cells.The energy of HCC cell was from glycolytic function,because the glycolysis was enhanced and the citric acid cycle decreased.Mitochondrial dysfunction and the increased cobalt content may correlate with the Warburg effect,which may be one of the pathogenesis of liver cancer,and expected to become the breakthrough point of a new targeting therapeutic approach.