1.Optimization of the Bombyx mori baculovirus expression system enhances the expression level of recombinant human keratinocyte growth factor-1 (hKGF-1).
Shuohao LI ; Xingyang WANG ; Xiaofeng WU ; Yujing XU ; Tian YANG ; Xinyu ZHU
Chinese Journal of Biotechnology 2025;41(7):2634-2646
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*
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Animals
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Baculoviridae/metabolism*
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Humans
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Fibroblast Growth Factor 7/biosynthesis*
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Recombinant Proteins/genetics*
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Cell Line
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Genetic Vectors/genetics*
2.The role of signaling crosstalk of microglia in hippocampus on progression of ageing and Alzheimer's disease
He LI ; Tianyuan YE ; Xingyang LIU ; Rui GUO ; Xiuzhao YANG ; Yangyi LI ; Dongmei QI ; Yihua WEI ; Yifan ZHU ; Lei WEN ; Xiaorui CHENG
Journal of Pharmaceutical Analysis 2023;13(7):788-805
Based on single-cell sequencing of the hippocampi of 5x familiar Alzheimer's disease(5x FAD)and wild type mice at 2-,12-,and 24-month of age,we found an increased percentage of microglia in aging and Alzheimer's disease(AD)mice.Blood brain barrier injury may also have contributed to this increase.Immune regulation by microglia plays a major role in the progression of aging and AD,according to the functions of 41 intersecting differentially expressed genes in microglia.Signaling crosstalk between C-C motif chemokine ligand(CCL)and major histocompatibility complex-1 bridges intercellular communi-cation in the hippocampus during aging and AD.The amyloid precursor protein(APP)and colony stimulating factor(CSF)signals drive 5x FAD to deviate from aging track to AD occurrence among intercellular communication in hippocampus.Microglia are involved in the progression of aging and AD can be divided into 10 functional types.The strength of the interaction among microglial subtypes weakened with aging,and the CCL and CSF signaling pathways were the fundamental bridge of communication among microglial subtypes.
3.Clinical Observation of Shenfu Injection Combined with Linezolid in the Treatment of Severe Pneumonia Caused by Methicillin-resistant Staphylococcus aureus
Jinsong LI ; Jijun CHEN ; Tao ZHU ; Xingyang WU
China Pharmacy 2017;28(20):2800-2803
OBJECTIVE:To observe clinical effect and safety of Shenfu injection combined with linezolid in the treatment of severe pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA). METHODS:In retrospective study,62 MRSA severe pneumonia patients selected from ICU of our hospital during Jun. 2012-Oct. 2015 were divided into observation group (25 cases)and control group(28 cases)according to medication plan. Based on routine treatment,control group was additionally given Linezolid injection 600 mg,ivgtt,bid. Observation group was additionally given Shenfu injection intravenously with initial dose of 60 mL,and then given continuous intravenous infusion of 60 mL at a rate of 20 mL/h,q12 h,on the basis of control group. Both groups were treated for consecutive 2 weeks. Clinical efficacy,bacteriological efficacy,cardiac function indexes and serum levels of inflammatory factors before and after treatment as well as the occurrence of ADR were compared between 2 groups. RESULTS:The clinical response rate of observation group was 88.00%,which was significantly higher than 60.71% of control group,with statistical significance (P<0.05). Bacterial clearance rate of observation group was 72.00%, and that of control group was 64.28%,there was no statistical significance between 2 groups(P>0.05). Before treatment,there was no statistical significance in cardiac function indexes and serum inflammatory factor levels between 2 groups(P>0.05). After treatment,LVEF,SV,CO and CI of 2 groups were increased significantly compared to before treatment,while the levels of TNF-α,IL-6,CRP and PCT were de-creased significantly;the indexes of observation group was significantly better than those of control group,with statistical signifi-cance (P<0.05). No severe ADR was found in 2 groups. CONCLUSIONS:Shenfu injection combined with linezolid effectively improves the cardiac function of patients with MRSA severe pneumonia,enhances the anti-inflammatory effect,and have a very significant clinic effect with good safety.
4.Periprosthetic femoral fracture after total knee arthroplasty
Xingyang ZHU ; Haitao SU ; Yongming HUANG
Chinese Journal of Tissue Engineering Research 2013;(39):6887-6895
BACKGROUND:Periprosthetic femoral fracture after total knee arthroplasty is related with the osteoporosis, bone defects, prosthesis, frail patients and high complication rate, so it is difficult to prevent and treat.
OBJECTIVE:To explore the risk factor, classification, treatment, rehabilitation and prophylaxis of periprosthetic femoral fracture after total knee arthroplasty based on the reviewed and summarized articles published in recent years.
METHODS:A computer-based online search was conducted in PubMed database from January 1, 1990 to December 31, 2011 and in SpringerLink database from 1980 to 2011 for the related articles with the key words of“periprosthetic fracture, knee”in English. A total of 626 articles were retrieved.
RESULTS AND CONCLUSION:According to inclusion and exclusion criteria, the articles were screened and 40 articles were included final y. The results showed that with the extensive development of total knee arthroplasty, the incidence of periprosthetic femoral fracture was increased gradual y;due to the poor prognosis, we should pay attention to the prevention. The risk factors of periprosthetic femoral fracture included patients’ internal factor that was hard to control, and some external factors such as the surgical techniques. Rorabeck classification was commonly applied for periprosthetic femoral fracture after total knee arthroplasty, but it was not perfect in clinical application. Kim classification wil be better for clinical guidance. The treatment of periprosthetic femoral fracture included nonoperative treatment, open reduction and internal fixation, retrograde intramedul ary nailing and revision arthroplasty. An appropriate treatment is chosen depending on fracture classification, local bone quality, patients’ medical and nutritional status. At present, however, there is not a perfect guideline for the selection of appropriate treatment method. But the early functional exercise is beneficial to prevent the related complications caused by longtime immobilization and the loss of joint function. Therefore, the indications must be under strict control in the treatment of periprosthetic femoral fracture after total knee arthroplasty. Except the firm fixation, early exercise for the patients should be encouraged at the same time.

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