Given the increasing concern regarding antibacterial resistance, the antimicrobial properties of naphthoquinones have recently attracted significant attention. While 1,4-naphthoquinone and its derivatives have been extensively studied, the antibacterial properties of 5,8-dihydroxy-1,4-naphthoquinone derivatives remain relatively unexplored. This study presents a comprehensive in vitro and in vivo analysis of the antibacterial activity of 35 naturally sourced and chemically synthesized derivatives of 5,8-dihydroxy-1,4-naphthoquinone. Kirby-Bauer antibiotic testing identified three compounds with activity against methicillin-resistant Staphylococcus aureus (MRSA), with one compound (PNP-02) demonstrating activity comparable to vancomycin in minimum inhibitory concentration, minimum bactericidal concentration (MBC), and time-kill assays. Microscopic and biochemical analyses revealed that PNP-02 adversely affects the cell wall and cell membrane of MRSA. Mechanistic investigations, including proteomic sequencing analyses, Western blotting, and RT-qPCR assays, indicated that PNP-02 compromises cell membrane integrity by inhibiting arginine biosynthesis and pyrimidine metabolism pathways, thereby increasing membrane permeability and inducing bacterial death. In an in vivo mouse model of skin wound healing, PNP-02 exhibited antibacterial efficacy similar to vancomycin. The compound demonstrated low toxicity to cultured human cells and in hemolysis assays and remained stable during serum incubation. These findings suggest that PNP-02 possesses promising bioactivity against MRSA and represents a potential novel antibacterial agent.
Methicillin-Resistant Staphylococcus aureus/genetics* ; Anti-Bacterial Agents/chemistry* ; Naphthoquinones/administration & dosage* ; Animals ; Microbial Sensitivity Tests ; Mice ; Humans ; Staphylococcal Infections/microbiology* ; Molecular Structure

Objective To explore the effect and mechanism of berberine combined with curcumin on ath-erosclerosis(AS)by mediating M1 macrophages polarization.Methods M1-type macrophages were obtained from mouse mononuclear macrophages(RAW264.7)induced by lipopolysaccharide(LPS,100 ng/mL)and interferon(IFN)-γ(20 ng/mL).A cell model was established.The cells were divided into a control group,model group,berberine group,curcumin group and berberine plus curcumin group.Concentrations of berberine and curcumin were detected by CCK-8 assay.The expression levels of M1-type macrophage markers iNOS,TNF-α,CXCL9 and p-STAT6/STAT6 in macrophage supernatant were detected by ELISA.Levels of iNOS,TNF-α and CXCL9 mRNA were detected by RT-PCR.Levels of iNOS,STAT6 and p-STAT6 proteins in each group were detected by Western blot.After down-regulation of STAT6 level by siRNA technology,expression of p-STAT6 protein was detected by Western blot.Expression levels of iNOS,TNF-α,CXCL9 and p-STAT6 were detected by ELISA.Results In the polarization of M1 macrophages induced by LPS and IFN-γ,berberine(25 μmol/L)and curcumin(20 μmol/L)were the best concentrations as compared with other drug concentration groups,and neither alone nor combined use could significantly inhibit the viability of RAW264.7 cells(P<0.05).As compared with the normal group,iNOS,TNF-α and CXCL9 mRNA and protein levels were increased in the model group,while P-STAT6/STAT6 levels were decreased,with statistical differences(P<0.05).As compared with the model group,iNOS,TNF-α and CXCL9 mRNA and protein levels in the berberine group,curcumin group,and berberine plus curcumin group were decreased,while P-STAT6/STAT6 levels were increased,and the changes were more obvious in berberine plus curcumin group,with statistical difference(P<0.05).After transfection of STAT6 siRNA in M1 macrophages in the berberine plus curcumin group,P-STAT6 levels were down-regulated,while expressions of iNOS,TNF-α and CXCL9 were up-regulated,with statistical differences(P<0.05).Conclusions Both berberine and curcumin can inhibit the activity of M1-type macrophages and reduce inflammatory response.The action of berberine combined with curcumin is more advantageous than that of either drug alone,which may be the main mechanism of action through activation of STAT6.

Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 μL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 ³ °C/min. The volume range of 1-8 μL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 ⁶ °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
Freezing ; Vitrification ; Cryopreservation/methods* ; Trehalose ; Gold ; Rewarming ; Metal Nanoparticles ; Cryoprotective Agents ; Lasers

Objective:To investigate the blood pressure change in patients with acute ischemic stroke (AIS) and hypertension treated with cinepazide maleate injection.Methods:This was a subgroup analysis of post-marketing clinical confirmation study of cinepazide maleate injection for acute ischemic stroke: a randomized, double-blinded, multicenter, placebo-parallel controlled trial, which conducted in China from August 2016 to February 2019. Eligible patients fulfilled the inclusive criteria of acute anterior circulation ischemic stroke with National Institutes of Health Stroke Scale (NIHSS) scores of 7-25. The primary endpoints were mean blood pressure of AIS patients treated with cinepazide maleate or control, which were assessed during the treatment period (14 days), and the proportion of the patients with normal blood pressure was analyzed after the treatment period. Furthermore, a subgroup analysis was performed to investigate a possible effect of the history of hypertension on outcomes.Results:This analysis included 809 patients with hypertension. There was no significant difference in patients blood pressure and the proportion of patients with normal blood pressure (60.5% vs. 59.0%, P>0.05) between cinepazide maleate group and control group. Conclusion:Administration of cinepazide maleate injection does not affect the management of clinical blood pressure in patients with AIS.

Genome-wide association studies (GWASs) are the most widely used method to identify genetic risk loci associated with orofacial clefts (OFC). However, despite the increasing size of cohort, GWASs are still insufficient to detect all the heritability, suggesting there are more associations under the current stringent statistical threshold. In this study, we obtained an integrated epigenomic dataset based on the chromatin conformation of a human oral epithelial cell line (HIOEC) using RNA-seq, ATAC-seq, H3K27ac ChIP-seq, and DLO Hi-C. Presumably, this epigenomic dataset could reveal the missing functional variants located in the oral epithelial cell active enhancers/promoters along with their risk target genes, despite relatively less-stringent statistical association with OFC. Taken a non-syndromic cleft palate only (NSCPO) GWAS data of the Chinese Han population as an example, 3664 SNPs that cannot reach the strict significance threshold were subjected to this functional identification pipeline. In total, 254 potential risk SNPs residing in active cis-regulatory elements interacting with 1 718 promoters of oral epithelium-expressed genes were screened. Gapped k-mer machine learning based on enhancers interacting with epithelium-expressed genes along with in vivo and in vitro reporter assays were employed as functional validation. Among all the potential SNPs, we chose and confirmed that the risk alleles of rs560789 and rs174570 reduced the epithelial-specific enhancer activity by preventing the binding of transcription factors related to epithelial development. In summary, we established chromatin conformation datasets of human oral epithelial cells and provided a framework for testing and understanding how regulatory variants impart risk for clefts.
Chromatin ; Cleft Lip/genetics* ; Cleft Palate/genetics* ; Epithelium ; Genome-Wide Association Study ; Humans

Objective To Investigate the cause of a food poisoning incident in a city in Dehong Prefecture， determine the scope of the incident， and to formulate effective prevention and control measures. Methods Field epidemiology and case-control study methods were used to formulate case definition； carry out case search； find suspicious food combined with clinical manifestations， dining history and other information； and collect samples from the suspicious food， cases and environment for laboratory testing. Results A total of 160 cases were found and the incidence rate was 26.02% （160/615）. The main clinical manifestations were diarrhea， leukocytosis， vomiting， nausea and abdominal pain. The average incubation period was 10.89±5.09 h， with 2 h as the shortest and 28 h as the longest. The epidemiological curve suggested the point source exposure mode. The results of case-control study showed that eating roast pork with green shoots was an independent risk factor （OR=13.09， 95%CI： 3.26‒52.54）.16 samples were tested by the local CDC laboratory. Proteus mirabilis was detected in two the anal swabs of two patients He and Chen. Both proteus pani and proteus mirabilis were detected in anal swab of patient Zhou and Proteus pani was detected in roast meat. There was no detection of pathogenic bacteria in other samples. Conclusion Based on the field epidemiological investigation and laboratory test results， we conclude that the food poisoning incident is caused by suspected proteus bacteria. We suggest that the market supervision department should strengthen the supervision of local food hygiene； clarify the sampling rights， responsibilities and technical procedures when medical institutions treat patients with foodborne diseases； ensure finding the causes in time； and take effective prevention and control measures to avoid similar food poisoning events.

Objective:To evaluate a new type of draw-bar skin stretcher in repair of full-thickness skin defects.Methods:From May 2015 to January 2019, 52 patients with full-thickness skin defects were repaired with a new type of draw-bar skin stretcher at Daping Hospital, Army Medical University. They were 40 males and 12 females, aged from 4 to 61 years (average, 37.1 years). Their skin was stretched for primary wound closure. When primary wound closure failed, skin stretching was performed again to close the wound depending on the wound condition. When the Pinch test was negative after skin stretching, the wound was sutured directly. In cases of positive Pinch test, a skin graft or flap was used to repair the remaining wound. At 12 months after surgery, scar contracture and size of skin graft or flap were observed and wound healing after skin stretching was evaluated in comparison with the original wound.Results:After skin stretching, one-stage wound closure was achieved in 36 cases and multi-stage wound closure in 8 cases; of the remaining 8 cases, 2 were repaired by skin graft and 6 by skin flap after their wounds were reduced by skin stretching. In one-stage closed wounds, infection occurred in 3 cases and marginal necrosis in 5 cases; in the wounds repaired by skin graft or flap, no infection or necrosis was observed. The 12-month follow-up for all the patients showed fine healing of all the wounds after one-stage or multi-stage closure, linear scar, absence of scar contracture, and smaller wound sizes than the original ones after skin graft or flap repair.Conclusions:Skin stretching using our new type of draw-bar skin stretcher is an effective treatment for skin wounds. It can replace traditional skin grafting and flap surgery in some cases, but its indications should be strictly followed to avoid related complications.

Objective To retrospectively analyze the clinical efficacy and safety of VCD (bortezomib/cyclophosphamide/dexamethason) in the treatment of multiple myeloma (MM) patients. Methods Fifty-five consecutive patients with newly diagnosed or relapsed MM were enrolled in the study retrospectively from June 2012 to June 2017. We collected and analyzed the clinical information of all patients treated with VCD.Results Firstly, the patients received therapy of VCD for median 4 cycles (Range : 2 ～8). The overall response rate (ORR) was 85.5％ (45/55). The complete response/near complete response rate (CR/nCR) , very good partial response rate (VGPR) and partial response rate (PR) were 27.3％, 23.6％ and 34.6％, respectively. The ORR of10 patients with renal inadequacy was 80.0％, while 45 cases with normal renal function was 82.2％ (P=0.627).Secondly, with a median follow-up of 13.5 months, the median progression free survival (PFS) , the median duration of response (DOR) and the median overall survival (OS) were 27 (1～61) months, 18 (1～50) months and 49 (1～64) months, respectively. Univariate prognostic analysis showed that abnormal ISS stage Ⅲ, relapse, renal dysfunction and response inferior to VGPR were negative prognostic factors for PFS, while abnormal ISS stageⅢ and renal dysfunction were negative prognostic factors for OS. Moreover, the multivariate prognostic analysis showed that abnormal ISS stage Ⅲ and response inferior to VGPR were independent prognostic factors for PFS, while ISS stage Ⅲ was independent prognostic factors for OS. Thirdly, the VCD treatment is effective and safe.The adverse events were evaluated according to International Myeloma Working Group Uniform Response Criteria.The results showed that the most common grade 3 ～4 non-hematology adverse events (AEs) were infection (20.0％) , peripheral neuropathy (5.5％) and hypertension (5.5％). The most common grade 3～4 hematology AEs were thrombocytopenia (10.9％) , neutropenia (9.0％) and anemia (5.5％). A total of 2 patients (3.6％) discontinued VCD because of serious peripheral neuropathy and 2 cases (3.65％) died of respiratory failure because of serious infection. Conclusions The VCD regimen is effective and safe in the treatment of newly diagnosed or relapsed/refractory MM patients in China. VCD is safe in patients with renal dysfunction.

Objective: To study the surgical strategy and clinical efficacy of hypertensive basal ganglia hematomas via transsylvian transinsular approach individually. Methods: The clinical data of 45 patients with hypertensive basal ganglia hematomas underwent microsurgical treatment with different sylvian anatomical points in Jianhu Hospital Affiliated to Nantong University from October 2014 to June 2016 were retrospectively analyzed. Results: The anterior hematomas was dissected through anterior point of lateral fissure, accounted for 66.7%(30 cases), the posterior hematoma was dissected through rolandic points under lateral fissure, accounted for 22.2%(10 cases), the long axis type hematoma was dissected between the anterior point of the lateral fissure and the lower rolandic point, accounted for 11.1%(5 cases). The postoperative CT scan showed that 42 cases were removed the hematomas for more than 90.0%, 3 cases were removed the hematomas for more than 75.0%, there was no postoperative rebleeding.According to GOS score, 14 cases returned to preoperative life status, 20 cases recovered sufficiently to return to family life, 9 cases could ambulate with a crotch but needed assistance, one case showed vegetative survival, one patient died. Conclusion Transsylvian transinsular approach via individual sylvian anatomical point should be advocated to remove basal ganglia hematomas, and it has the advantages of minimally invasion, high hematoma evacuation rate, low rebleeding rate, good neurological recovery and so on.

Objective: To investigate the effect of papaverine on wound healing of full-thickness skin defect after skin stretching in swine. Methods: Eight Bama pigs were prepared. Standard full-thickness skin defect for 7 cm×7 cm was produced in the middle part of each pig's forelimb, and skin traction was used to close the wound. According to random number table method, the pigs were divided into experiment group and control group, with four pigs in each group. Percutaneous oxygen partial pressure (TcPO₂) was compared preoperatively and postoperatively. Wound healing clinical score was evaluated four and 10 days after operation. Weidner counting method was employed to determine the microvascular density (MVD) of the wound edge tissue 0 and 10 days after operation. Immunohistochemistry was used to observe the expression of hypoxia inducible factor 1-α (HIF- 1α) and vascular endothelial growth factor (VEGF) 0, 4 and 10 days after operation. Results: There was no statistically significant difference in TcPO₂ between the experiment group [(55.1±5.4)mmHg] and the control group [(54.7±5.9)mmHg] before skin stretching (P>0.05), while statistically significant difference was found after stretching [(22.7±3.3)mmHg vs. (16.4±3.4)mmHg] (P<0.01). There was no statistically significant difference in clinical wound healing scores between the control group and experiment group 4 days after skin stretching [(2.6±0.7)points vs. (2.1±0.4)points] (P>0.05), but better result was found in experiment group compared with the control group in 10 days [(1.3±0.5)points vs. (4.6±0.7)points] (P<0.01). MVD of skin tissue showed no statistically significant difference between the control group (8.5±1.3) and the experiment group (9.3±1.2) immediately after skin stretching (P>0.05), while the MVD of wound skin tissue in the experiment group (29.5±4.8) was significantly higher than that in the control group (21.1±3.4) 10 days after skin stretching (P<0.01). After skin stretching, the HIF-1 alpha expression in experiment group was significantly higher than the control group 4 days after skin stretching [(50.0±7.0) vs. (38.6±7.0)] (P<0.01), but no statistical significance was found between that at 0 day and 10 days(P>0.05). The expression of VEGF in the wound skin tissues of the experiment group was significantly higher than that of the control group [(49.9±10.8) vs. (39.2±6.3)] at 4 days after skin stretching (P<0.05), while no statistically significance was found at 0 and 10 days (P>0.05). Conclusion For full-thickness skin defect after skin stretching in pigs, papaverine can alleviate small vessel spasm, improve skin edge oxygen supply, and promote the growth of microvessels, thus promoting the healing, which might be associated with the increase of HIF-1ɑ and VEGF expression.