Objective To explore the mechanism of the formation of glioma cells derived vessels (GCDV).Methods The tubular formation assay was performed on 3D cell cultures of U87,U251,U373,SF295,T98G,SKMG-4 and C6 glioma cell lines.The expression of Notch1 、Dll4 were examined by western blot analysis.Result The mean number of vasculogenic channels of cell lines C6,U373,U87,T98G,SF295,U251 and SKMG-4 per area (100×) were 25.2±5.0,36.4±3.2,19.0±2.2,12.6±2.4,4±2.2,0.2±0.4 and 0,respectively.The relative protein densities of Notchl in U87,U251,T98G,SF295,SKMG-4,C6 and U373 were 0.34,0.21,0.79,0.04,0.28,1.75 and 1.19,which were significantly related with the tubular formation ability (P＜0.05).However,the expression of Dll4 was not associated with tubular formation ability (P＞0.05).Conclusion Notch1 may play a key role in tubular formation of glioma cells while the role of Dll4 in the process needs further study.

To study the chemical constituents of Veratrum dahuricum (Turcz.) Loes. f., a new aurone glycoside named as (Z)-7, 4'-dimethoxy-6-hydroxyl-aurone-4-O-β-glucopyranoside was isolated from the 95% ethanol extracts of the rhizomes and roots of Veratrum dahuricum (Turcz.) Loes. f. by repeated column chromatography on silica gel and recrystallization. Its structure was established by extensive spectroscopic analyses, and its cytotoxicities against HepG-2, MCF7 and A549 cell lines were measured in vitro.

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 < 25.0 micromo x L(-1)) than that of parent ciprofloxacin (IC50 > 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.

Objective To explore the influences of extracellular matrices (ECM) secreted by ultraviolet B (UVB)-induced senescent fibroblasts on the proliferation of and extracellular signal-regulated kinase (ERK) signaling in HaCaT cells. Methods Fibroblasts were irradiated with UVB of 15 mJ/cm2 once daily for 5 days to induce premature senescence, which was identified by SA-β-gal staining 72 hours after the last irradiation.HaCaT cells were divided into 3 groups and inoculated into plates coated with extracellular matrices secreted by non-senescent (PRE-ECM) or senescent fibroblasts (SIPS-ECM) or into uncoated plates (NON-ECM), fol-lowed by additional culture. U0126, an inhibitor of ERK1/2, was used to treat the HaCaT cells 1 hour before inoculation. Then, MTT assay was carried out to detect the proliferation of HaCaT cells after a 3-day culture,Western blot to assess the phosphorylation of ERK at 0.5, 1, 2 and 4 hours after the inoculation, flow cytometry to analyse cell cycle and apoptosis after 24 hours of culture. Results The most rapid and intense phosphory-lation of ERK was observed in SIPS-ECM group. Inhibiting the activation of ERK pathway with U0126 could completely suppress the promoting effect of ECM from senescent fibroblasts on the proliferation of HaCaT cells.After the blocking of ERK activation, the proportion of HaCaT cells in S and G2/M phase decreased from 37.40%, 41.34% and 43.31% to 29.41%, 36.48% and 39.96%, respectively, in NON-ECM, PRE-ECM and SCIP-ECM group. Conclusion The ECM produced by UVB-induced senescent fibroblasts promote the prolifera-tion of HaCaT cells via inducing the phosphorylation of ERK.

Objective To investigate the risk factors of perioperative heart failure in the elderly patients with femoral neck fracture treated with hip joint replacement so as to provide relevant clinical reference.Methods A retrospective case control study was conducted to analyze the clinical data of 155 elderly patients with femoral neck fractures who underwent total hip or hemiarthroplasty in Wenzhou Traditional Chinese and Western Medicine Hospital from May 2012 to August 2016.There were 62 males and 93 females,aged (75.6 ± 7.4)years.The patients were divided into heart failure group (26 patients)and non heart failure group (129 patients).Heart failure group included 10 males and 16 females,aged (78.3 ± 8.2)years.There were 52 males and 77 females in the non heart failure group,aged (74.5 ±6.7) years.Information including age,gender,history of hypertension,history of heart diseases,American Society of Anesthesiologists (ASA) physical status classification,duration from injury to surgery,preoperative renal function,perioperative fluid balance,operation method,operation time,postoperative pain score,perioperative blood loss,and constipation were collected.Univariate analysis was firstly performed on the above data,and multivariate logistic regression analysis was conducted on the significant factors generated by the univariate analysis so as to identify independent risk factors for perioperative heart failure.Results According to the univariate analysis,age,history of heart diseases,preoperative renal function,and perioperative fluid balance were statistically different between the two groups (P ＜ 0.05).Multivariate analysis results showed that the independent risk factors of perioperative heart failure included age (OR =5.351,95％ CI 1.459-19.623,P ＜ 0.01),history of heart diseases (OR =5.717,95 ％ CI 2.399-13.624,P ＜ 0.01),preoperative renal function (OR =2.670,95％ CI 1.125-6.336,P ＜ 0.05),perioperative fluid balance (OR =2.980,95％ CI 1.287-6.899,P ＜0.01).Conclusions Age,history of heart diseases,preoperative renal function,and perioperative fluid balance are the risk factors of perioperative heart failure in elderly patients with femoral neck fracture.Therefore,more attention should be paid to these risk factors and corresponding preventative measures should be taken to reduce the incidence of perioperative heart failure.

The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice. Two 6-mm wounds along the both sides of the spine were created on the back of each mouse (n=80) by using a punch biopsy. The mice were assigned randomly into two groups, with 40 animals in each group: a non-thermal plasma group in which the mice were treated with the non-thermal plasma; a control group in which the mice were left to heal naturally. Wound healing was evaluated on postoperative days (POD) 4, 7, 10 and 14 (n=5 per group in each POD) by percentage of wound closure. The mice was euthanized on POD 1, 4, 7, 10, 14, 21, 28 and 35 (n=1 in each POD). The wounds were removed, routinely fixed, paraffin-embedded, sectioned and HE-stained. A modified scoring system was used to evaluate the wounds. The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group, earlier than in control group in which acute inflammation reached a peak on POD 7, and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7 (P<0.05). The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group (P<0.05). The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10 (P<0.05 for all). The count of bacterial colonies was 10(3) CFU/mL on POD 4 and <20 CFU/mL on POD 7, significantly lower than that in control group (10(9) CFU/mL on POD 4 and >10(12) CFU/mL on the POD 7) (P<0.05). It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colonization.

Objective:To observe the effects of proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor combined with rosuvastatin on lipid levels and short-term prognosis in patients with acute ischemic stroke, and to explore the optimal therapeutic regimen in terms of efficacy and safety, so as to provide a basis for clinical practice.Methods:Consecutive patients with acute cerebral infarction within 72 hours of onset and lipids≥2.6 mmol/L admitted to Xuzhou Central Hospital from April 2022 to March 2023 were included in the study, and the randomized numeric table method was used to divide them into 3 groups of different treatment regimens, group A (rosuvastatin 20 mg, once a day), group B (rosuvastatin 10 mg, once a day+alirocumab 75 mg, once 2 weeks) and group C (rosuvastatin 20 mg, once a day+alirocumab 75 mg, once 2 weeks). General baseline data, National Institutes of Health Stroke Scale score, modified Rankin Scale score on day 90 and the occurrence of adverse events and serious adverse events were collected from the 3 groups. The primary efficacy outcome was the degree of reduction in low density lipoprotein-cholesterol (LDL-C) from baseline on day 90. The secondary efficacy outcomes were recurrence rate and time to recurrence in stroke patients within 90 days,etc. The primary safety outcome was hepatic insufficiency (transaminase elevation≥3 times normal) within 90 days. The secondary safety outcomes were death due to stroke within 90 days and fatal and nonfatal myocardial infarction.Results:A total of 501 patients were included, 166 patients in group A, 167 patients in group B, and 168 patients in group C. The differences in the baseline data of the 3 groups were not statistically significant (all P>0.05). LDL-C was reduced from baseline on day 90 in groups A, B, and C, with the differences of -1.5 (-1.7, -1.4) mmol/L, -2.2 (-2.5, -2.1) mmol/L and -2.2 (-2.6, -2.1) mmol/L, respectively, with statistically significant differences among the 3 groups ( H=1.497, P<0.001); the differences between the group A and group B, and between the group A and group C, were statistically significant ( Z=-11.125, P=0.006; Z=-9.475, P=0.012), while the difference between the group B and group C was not statistically significant ( Z=1.650, P=0.946). The numbers of 90-day stroke recurrence cases (recurrence rate) in patients in the groups A, B, and C were 12 (7.2%), 4 (2.4%), and 5 (3.0%), respectively, without statistically significant difference among the 3 groups ( χ 2=5.773, P>0.05); the recurrence time of patients in the groups A, B and C was (43.0±7.4) d, (66.0±8.3) d and (62.2±5.6) d, respectively, and the differences among the 3 groups were statistically significant ( F=14.096, P=0.001). Compared with the group A, patients in the groups B and C had a prolonged time to stroke recurrence within 90 days ( Z=-3.108, P=0.002; Z=-2.871, P=0.004), whereas the difference in the time to stroke recurrence within 90 days between patients in the groups B and C was not statistically significant ( Z=0.397, P=0.692). The time to stroke recurrence within 90 days was positively correlated with the level of LDL-C on day 90 ( β=0.850, P=0.031). Ten patients (6.0%) in the group A developed hepatic insufficiency, 1 patient (0.6%) in the group B, and 9 patients (5.4%) in the group C. The differences among the 3 groups were statistically significant (χ 2=7.622, P=0.022); and the difference between the group B and group C was statistically significant ( P=0.011). The differences of secondary safety endpoints, death due to stroke within 90 days [1 case (0.6%) in the group A, 0 case (0) in the group B, and 1 case (0.6%) in the group C], fatal and nonfatal myocardial infarction within 90 days [3 cases (1.8%) in the group A, 1 case (0.6%) in the group B, and 1 case (0.6%) in the group C], were not statistically significant among the 3 groups (all P>0.05). Conclusions:In patients with acute ischemic stroke, PCSK9 inhibitor combined with rosuvastatin (both medium and high doses) significantly reduced LDL-C levels compared with baseline, and at the same time prolonged the time to stroke recurrence, reduced adverse effects such as hepatic insufficiency, and had a high degree of safety. PCSK9 inhibitor combined with medium-dose rosuvastatin had a better effect.

The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice.Two 6-mm wounds along the both sides of the spine were created on the back of each mouse (n=80) by using a punch biopsy.The mice were assigned randomly into two groups,with 40animals in each group:a non-thermal plasma group in which the mice were treated with the non-thermal plasma; a control group in which the mice were left to heal naturally.Wound healing was evaluated on postoperative days (POD) 4,7,10 and 14 (n=5 per group in each POD) by percentage of wound closure.The mice was euthanized on POD 1,4,7,10,14,21,28 and 35 (n=1 in each POD).The wounds were removed,routinely fixed,paraffin-embedded,sectioned and HE-stained.A modified scoring system was used to evaluate the wounds.The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group,earlier than in control group in which acute inflammation reached a peak on POD 7,and the acute inflammation scores were much lower in non-thermal group than in control group on POD7 (P＜0.05).The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group (P＜0.05).The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10 (P＜0.05 for all).The count of bacterial colonies was 103 CFU/mL on POD 4 and ＜20 CFU/mL on POD 7,significantly lower than that in control group (109 CFU/mL on POD 4 and ＞1012 CFU/mL on the POD 7) (P＜0.05).It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colonization.

Objective:To establish and identify a SD rat model with reconstruction of bladder sensory and motor innervation , so as to lay the foundation for further study of micturition center remodeling and its mechanisms .Methods:A total of 45 female SD rats were randomly divided into control group(n=10) ,rhizotomy group(n=15) and nerve root anastomosis group(n=20) . All the ventral and dorsal roots of spinal nerve below L4 level of rats in rhizotomy group were cut off .In nerve root anastomosis group ,the bilateral ventral and dorsal roots of L4 nerve ,were anastomosed with those of S1 nerve ,after the spinal nerve roots had been cut off .Rats in control group were not treated with surgery . At Six months after surgery ,rats in each group underwent urodynamic test ,nerve root stimulation ,toluidine blue staining at nerve anastomosis site ,pelvic ganglia fluorescence gold tracer staining and bladder wet weight measurements .Results:Bladder maximum capacity ,residual urine volume ,bladder compliance and bladder wet weight in nerve root anastomosis group was less than those in rhizotomy group ,however ,larger than those in control group(P<0 .05) .There was no statistically significant difference in maximum voiding pressure between nerve root anastomosis group and control group(P> 0 .05) ,however ,maximum voiding pressure in nerve root anastomosis group was larger than that in rhizotomy group(P<0 .05) .Intravesical pressure increased after nerve root stimulation in nerve root anastomosis group ,but it was still lower than that in control group(P<0 .05) .Nerve passing rate was (53 .4 ± 6 .7)% in nerve root anastomosis group ,under toluidine blue staining at nerve anastomosis site .After injection of fluorescent gold in pelvic ganglia ,fluorescence gold staining was visible in the L4 spinal cord gray matter in nerve root anastomosis group , however ,not visible in rhizotomy group and control group .Conclusions:SD rat model with reconstruction of bladder sensory and motor innervation is successfully established .It lays the foundation for further study of micturition center remodeling and its mechanism .

Objective:Tumor-treating fields (TTFields) is a kind of non-invasive anti-mitotic tumor therapy, which has been approved for patients with newly diagnosed and recurrent glioblastoma. This study aims to explore the efficacy and safety of TTFields in high-grade gliomas in clinical practice settings.Methods:The clinical data of 15 patients with recurrent glioma and 9 patients with newly diagnosed high-grade glioma admitted to our center from April 2019 to January 2021 were retrospectively analyzed. All patients accepted TTFields≥1 month. Follow-up was performed for 5.3 months (ranged from 2.3 to 10.7 months); Response Assessment in Neuro-Oncology Working Group (RANO) criteria was used to evaluate the glioma responses. The progression-free survival (PFS) and overall survival (OS) were calculated according to Kaplan-Meier method. Common Terminology Criteria for Adverse Events v5.0 (CTCAE v5.0) and TTFields related skin adverse reaction (dAE) criteria were used to evaluate the adverse events. Quality of life questionnaire-core 30 (QLQ-C30) and QLQ-brain cancer module (QLQ-BN20) questionnaires were used to evaluate the health-related quality of life (HRQoL). Treatment compliance was evaluated by data on the use of NovoTTF-200A devices, and calculated as a percentage of daily TTFields usage.Results:The median duration of TTFields was 4.2 months (ranged from 1.0 to 10.7 months), with a median compliance rate of 91.5% (67.0%-97.0%). TTFields was used alone in 2 patients and used with combination of chemotherapy in 22 patients. From follow-up to April 2021, 14 patients had stable symptoms and 10 had disease progression (8 died). The median PFS and OS of recurrent patients were 5.9 months ( 95%CI: 3.3-8.6 months) and 8.5 months ( 95%CI: 3.2-13.8 months), respectively; and the median PFS and OS of newly diagnosed patients were both 10.7 months (without 95%CI). The common adverse events included grading 1 dAE (58.3%) and grading 2 dAE (12.5%), without grading 3 or 4 dAE, manifested as contact or allergic dermatitis, erosion, folliculitis and ulcers. And 87.5% patients had stable HRQoL. Conclusions:The preliminary results showed that the survival of recurrent high-grade glioma patients treated by TTFields is similar to that reported in foreign literature; and the newly diagnosed patients need further survival follow-up. The patients' treatment compliance and safety are good. The dAE incidence (grading 1-2) is higher than that reported in the literature, and the toxicity was acceptable.