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The emergence of carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) has driven us to explore alternative treatments for the limitation of antimicrobial agents. Lytic phages are considered a promising alternative treatment for CR-hvKP infection. In this study, we reported three novel lytic phages, vB_KpnA_SCNJ1-Z, vB_KpnS_SCNJ1-C, and vB_KpnM_SCNJ1-Y, against a CR-hvKP strain SCNJ1, and they possess genomes of double-stranded DNA with a size of 43,428 bp, 46,039 bp, and 50,360 bp, respectively. Phylogenetic analysis demonstrated that vB_KpnA_SCNJ1-Z belongs to the family Autographiviridae within the class Caudoviricetes, while vB_KpnS_SCNJ1-C and vB_KpnM_SCNJ1-Y are unclassified Caudoviricetes. The phages showed a narrow host range only lysing 1 of 50 tested clinical bacterial strains. The one-step growth curves and stability results showed that the phages displayed relatively short latency periods, with broad pH (pH 3-14) and thermal stabilities (20–60°C). The phages showed significant inhibition of the biofilm formation by SCNJ1 and strong antibacterial activity in vitro. In the mouse model, we demonstrated that administration of a single phage or phage cocktail significantly reduced bacteria loads in the lung, liver, and spleen, and effectively rescued mice from the infection of the SCNJ1 strain, with a survival rate of 70-80%. These findings suggested the three phages have great potential as an alternative therapy with favorable stability and strong antibacterial activity both in vivo and in vitro for the treatment of CR-hvKP infection.

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a powerful gene editing tool originated from prokaryotes. The modern CRISPR/Cas9 system allows high-throughput genetic screening to be carried out in vitro and in vivo. The high efficacy and flexibility of CRISPR/Cas9 system allows identification of hepatocellular carcinoma (HCC) related genes in the past decades. Numerous efforts have been applied in the past to improve this system, such as off-target improvement, gRNA efficiency enhancement, and modified Cas9 nuclease for performing visionary base editor screens and epigenetic screens. With these merits, the CRISPR/Cas9 offers tremendous opportunities in various biomedical research and clinical application. Recently, the use of the CRISPR/Cas9 system has also been combined with different technologies, including single-cell sequencing and machine learning, to further understand HCC pathogenesis and explore its utility in gene therapy. This review provides a summary of HCC carcinogenesis CRISPR/Cas9 screens conducted in recent years with different genetic contexts, epidemiological backgrounds, and progression of HCC. Furthermore, this review also provides insight in the CRISPR/Cas9 potentials, current obstacles, and improvement of this system for its future utility in cocktail therapies.

Abstract Background Mono-therapeutic modality has limitations in combating metastatic lesions with complications. Although emerging immunotherapy exhibits preliminary success, solid tumors are usually immunosuppressive, leading to ineffective antitumor immune responses and immunotherapeutic resistance. The rational combination of several therapeutic modalities may potentially become a new therapeutic strategy to effectively combat cancer. Results Poly lactic-co-glycolic acid (PLGA, 50 mg) nanospheres were constructed with photothermal transduction agents (PTAs)-Prussian blue (PB, 2.98 mg) encapsulated in the core and chemotherapeutic docetaxel (DTX, 4.18 mg)/ immune adjuvant-imiquimod (R837, 1.57 mg) loaded in the shell. Tumor cell membranes were further coated outside PLGA nanospheres (designated “M@P-PDR”), which acted as “Nano-targeted cells” to actively accumulate in tumor sites, and were guided/monitored by photoacoustic (PA)/ magnetic resonance (MR) imaging. Upon laser irradiation, photothermal effects were triggered. Combined with DTX, PTT induced in situ tumor eradication. Assisted by the immune adjuvant R837, the maturation rate of DCs increased by 4.34-fold compared with that of the control. In addition, DTX polarized M2-phenotype tumor-associated macrophages (TAMs) to M1-phenotype, relieving the immunosuppressive TME. The proportion of M2-TAMs decreased from 68.57% to 32.80%, and the proportion of M1-TAMs increased from 37.02% to 70.81%. Integrating the above processes, the infiltration of cytotoxic T lymphocytes (CTLs) increased from 17.33% (control) to 35.5%. Primary tumors and metastasis were significantly inhibited when treated with “Nano-targeted cells”-based cocktail therapy. Conclusion “Nano-targeted cells”-based therapeutic cocktail therapy is a promising approach to promote tumor regression and counter metastasis/recurrence. Graphical Abstract

Objective To analyze the results of unicompartmental knee arthroplasty (UKA) in patients with single compartment of knee osteoarthritis, and to evaluate the clinical outcomes and safety with different analgesias. Methods Totally 45 cases were recruited from January 2017 to December 2019 including 18 males and 27 females. The mean age was (63.41±1.19) year-old. They were divided into three groups with 15 in each.Patients in group A received cocktail treatment after surgery. Patients in group B received adductor canal blockade treatment by ultrasound guide after surgery. and those in group C received both cocktail treatment and adductor canal blockade treatment by ultrasound guide after surgery. VAS score and KSS score were compared and analyzed according to postoperative follow-up results. Results VAS scores and KSS scores of the three groups before surgery were not different but VAS scores and KSS scores of the two groups were significantly reduced after treatment (P＜0.05). Post-operative VAS score and KSS score were significantly different between the two groups (P＜0.05). Conclusions As an effective final treatment for single compartment of knee osteoarthritis, UKA is the most effective treatment in the early and middle stages of knee preserving surgery. Combining cocktail treatment and adductor canal blockade treatment by ultrasound guide after surgery can provide a safe post-operative analgesia for unicompartmental knee arthroplasty.

Abstract Background Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), an RNA virus with a high mutation rate. Importantly, several currently circulating SARS-CoV-2 variants are associated with loss of efficacy for both vaccines and neutralizing antibodies. Methods We analyzed the binding activity of six highly potent antibodies to the spike proteins of SARS-CoV-2 variants, assessed their neutralizing abilities with pseudovirus and authentic SARS-CoV-2 variants and evaluate efficacy of antibody cocktail in Delta SARS-CoV-2-infected hamster models as prophylactic and post-infection treatments. Results The tested RBD-chAbs, except RBD-chAb-25, maintained binding ability to spike proteins from SARS-CoV-2 variants. However, only RBD-chAb-45 and -51 retained neutralizing activities; RBD-chAb-1, -15, -25 and -28 exhibited diminished neutralization for all SARS-CoV-2 variants. Notably, several cocktails of our antibodies showed low IC50 values (3.35–27.06 ng/ml) against the SARS-CoV-2 variant pseudoviruses including United Kingdom variant B.1.1.7 (Alpha), South Africa variant B.1.351 (Beta), Brazil variant P1 (Gamma), California variant B.1.429 (Epsilon), New York variant B.1.526 (Iota), and India variants, B.1.617.1 (Kappa) and B.1.617.2 (Delta). RBD-chAb-45, and -51 showed PRNT50 values 4.93–37.54 ng/ml when used as single treatments or in combination with RBD-chAb-15 or -28, according to plaque assays with authentic Alpha, Gamma and Delta SARS-CoV-2 variants. Furthermore, the antibody cocktail of RBD-chAb-15 and -45 exhibited potent prophylactic and therapeutic effects in Delta SARS-CoV-2 variant-infected hamsters. Conclusions The cocktail of RBD-chAbs exhibited potent neutralizing activities against SARS-CoV-2 variants. These antibody cocktails are highly promising candidate tools for controlling new SARS-CoV-2 variants, including Delta.

The Streptococcus mutans (S. mutans) phage, as one of the principal pathogenic bacteria of dental caries, is a main cause of the formation and development of dental caries due to its overproliferation in dental plaque biofilms. Bacterial viruses, also known as bacteriophages, have the capability of specifically infecting bacteria and effectively degrading bacterial biofilms. S. mutans phages, therefore, may prevent and control caries. Therapy based on phages has been applied in many fields, but the application of S. mutans phages in caries remains exploratory. This article will review the research progress of S. mutans phages in clinical caries prevention, aiming to provide a new idea for the clinical prevention of caries. The results of the literature review show that the living bacteriophage system has the advantages of high specificity, high affinity and good safety. However, due to its unstable structure, it can be processed into a more stable formulation by freeze-drying, spray drying, adding stability enhancers, or incorporating bacteriophages into ointments, biodegradable polymer matrices or particles to a certain extent to improve stability. The lysozyme produced by phages can digest the bacterial cell wall and release the assembled phage particles, which effectively cleave biofilms. In addition, the antigen binding fragment library for cariogenic pathogens was screened by phage display technology, and the purpose of caries prevention and treatment was achieved by passive immunization of antigen binding fragments. However, the host range of bacteriophages is narrow, so this kind of problem can be overcome by phage combined with traditional therapy or other drug use or cocktail therapy with multiple phages in clinical caries prevention and control.

Background The treatment of keloids is both challenging and controversial. Until now, there is no definitive treatment modality for keloids. To determine the efficacy and safety of an intralesional cocktail therapy, including bleomycin compared to corticosteroid injection method on the treatment of keloid scars. Methods A retrospective chart review was performed for 35 patients diagnosed with keloids. The cocktail therapy regimen consisted of bleomycin, 1% lidocaine, verapamil, and triamcinolone, which was further diluted with 1 mL of normal saline. In group B, 1mL of triamcinolone acetonide was injected into the lesion. Results for a total of 35 patients included in this study were evaluated according to their therapeutic responses and complications six months after the last session. Results Analyzing the outcomes of the patients treated with a cocktail regimen (group A), it was found that 87% showed favorable therapeutic responses (scar thickness

CD19 chimeric antigen receptor T-cell (CAR-T) therapy is efficacious for refractory/relapsed (R/R) B-cell hematological malignancies, yet relapse due to CD19 antigen escape remains a challenge. Our trial explored simultaneous targeting of multiple B-cell antigens as a therapeutic approach that may reduce the risk of relapse. We tested the safety and efficacy of CAR19/22 T-cell cocktail therapy including murinized and humanized products among patients with R/R aggressive B-cell lymphoma. In the group that received the humanized product, 11/12 (91.7%) patients achieved an objective response, including 9/12 (75%) complete responses (CRs) by day 28. The overall response rate and CR rate in the murinized group was 92.9% (13/14) and 42.9% (6/14), respectively. Nine of 12 (75%) patients in the humanized group maintained CR at month 3 following infusion, compared to 5/14 patients (35.7%) in the murinized group. Progression-free survival (PFS) was more favorable in the humanized compared to the murinized group. Most patients had mild cytokine release syndrome (CRS) (grade 1–2) in both groups. This study demonstrates that CAR19/22 T-cell cocktail therapy is safe and effective for R/R B-cell lymphoma and that patients treated with a humanized CAR-T exhibited better efficacy compared to patients treated with a murinized CAR-T therapy.

Chenhao Zhang,1 Yanjing Zang,2 Qin Song,3 Hongxuan Li,1 Lei Hu,1 Weidong Zhao,3 Shanshan Feng,1 Fang Gu,4 FengLi Zhao,1 Chunliang Zhang11Department of Neurology, The Second Hospital of Baoding City, Baoding, Hebei 071051, People’s Republic of China; 2Department of Geriatric, The Second Hospital of Baoding City, Baoding, Hebei 071051, People’s Republic of China; 3Second Neurology Department, The Second Hospital of Baoding City, Baoding, Hebei, 071051, People’s Republic of China; 4Fifth Department of Internal Medicine, Baoding Children’s Hospital, Baoding, Hebei 071051, People’s Republic of ChinaObjective: To explore the efficacy and safety of “cocktail therapy” in Parkinson’s disease with dementia (PDD).Patients and methods: Sixty patients with PDD were randomly assigned to the test group (n=30) and a control group (n=30). The control group received 10 mg of donepezil hydrochloride tablets orally, once a day. The test group was treated with a “cocktail therapy”, which consisted of 10 mg of donepezil hydrochloride tablets taken orally, once a day; 200 mg of dl-3n-butylphthalide soft capsules taken orally, 15 mins before each meal, three times daily; 800 mg of oxiracetam capsules taken orally, three times daily; and 80 mg of Ginkgo biloba extract tablets taken orally, three times daily. Treatment was administered for six months. The Montreal cognitive assessment scale (MoCA), Blessed-Roth dementia scale, and Clinical Dementia Rating Scale sum of boxes (CDR-SB) were used before treatment and on the third and sixth month after treatment. Adverse events were also monitored.Results: There were no statistical differences in MoCA, CDR-SB, or Blessed-Roth dementia scale scores between the two groups before treatment. MoCA scores of the test group at six months were significantly higher than those before the treatment and at three months, while both the Blessed-Roth and CDR-SB scores were significantly lower than those before treatment and at three months (p0.05) between the test group (16.67%) and the control group (13.33%) in the rate of abnormal liver function after treatment.Conclusion: Treatment with “cocktail therapy” was safe and improved the conditions of patients with PDD, as well as the quality of life of the patients.Keywords: cocktail therapy, Parkinson’s disease with dementia, efficacy, donepezil hydrochloride, cocktail therapy

Huanglongbing (HLB) is a destructive citrus bacterial disease caused by Candidatus Liberibacter asiaticus (Ca.Las) and cannot be cured by current pesticides. Root lesion and Tylenchulus semipenetrans juveniles were observed in HLB-affected citrus tree roots. We hypothesize that root treatment with fosthiazate (FOS) and Cupric-Ammonium Complex (CAC) will improve the root growth and inhibit HLB. CAC is a broad spectrum fungicide and can promote growth of crops. FOS kills Tylenchulus semipenetrans and protects roots from damage by harmful bacteria such as Ca.Las. After 90 days of combination treatment of FOS and CAC through root drenches, the citrus grew new roots and its leaves changed their color to green. The inhibition rate of Ca.Las reached more than 90%. During treatment process, the chlorophyll content and the root vitality increased 396 and 151%, respectively, and starch accumulation decreased by 88%. Transmission electron microscopy (TEM) and plant tissue dyeing experiments showed that more irregular swollen starch granules existed in the chloroplast thylakoid system of the HLB-infected leaves. This is due to the blocking of their secretory tissue by starch. TEM and flow cytometry experiments in vitro showed the synergistic effects of FOS and CAC. A transcriptome analysis revealed that the treatment induced the differential expression of the genes which involved 103 metabolic pathways. These results suggested that the cocktail treatment of FOS and CAC may effectively kill various pathogens including Ca.Las on citrus root and thus effectively control HLB.