[Objective] To investigate the m vitro effect of asiaticoside on the proliferation of and connective tissue growth factor (CTGF) expression by keloid-derived fibroblasts.[Methods] Tissue samples from patients with keloid were obtained for primary culture of fibroblasts.After 3 to 7 passages,the fibroblasts were incubated with different concentrations (100,10,1,0.1,0.01 mg/L) of asiaticoside or dimethyl sulfoxide (DMSO) for 24,48 and 72 hours followed by the determination of cell viability by methyl thiazolyl tetrazolium (MTT) assay.Immunohistochemistry and Western blot were carried out to quantify the expression of CTGF in the fibroblasts at 48 hours after treatment with different concentrations of asiaticoside.The morphology of fibroblasts was observed before and after the treatment with asiatieoside.[Results] As morphological observation showed,different concentrations of asiaticoside induced an obvious apoptosis and growth inhibition in fibroblasts.The growth of fibroblasts was suppressed by asiaticoside of 1-100 mg/L in a dose-dependent manner (r =0.95,0.90,0.92 for 24-,48- and 72-hour treatment respectively,all P ＜ 0.01 ),and one-factor analysis of variance revealed statistical differences in the growth inhibition rate in fibroblasts between different treatment durations for each tested concentration of asiaticoside (all P ＜ 0.01 ).There was a strong expression of CTGF in untreated fibroblasts,which was weakened by the treatment with asiaticoside for 48 hours.The number of CTGF-positive fibroblasts per 100 cells was 73 in untreated fibroblasts,significantly higher than that in fibroblasts treated with asiaticoside at 1 mg/L (54,t =4.34,P ＜ 0.01 ) and 10 mg/L (46,t =6.26,P ＜ 0.01 ),and statistical differences were observed between the fibroblasts treated with asiaticoside at 1 mg/L and 10 mg/L (t =1.95,P ＜ 0.05).Western blot also showed that the expression of CTGF was inhibited by 48-hour treatment with asiaticoside,and the inhibitive effect displayed a trend to increase with the rise in the concentration of asiaticoside.[Conclusion] Asiaticoside can effectively inhibit the oroliferation of and CTGF expression by fibroblasts in vitro.

Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.

Objective To investigate the efficacy and the indication and the management of perioperative complications in treatment of infra- kidney abdominal aortic aneurysm (AAA) by using endovascular graft exclusion (EVGE). Methods From April 2006 to September 2008, 24 patients with infra- kidney abdominal aortic aneurysms were diagnosed by contrast-enhanced CT or MRI scan. Vascular access was obtained through the bilateral femoral artery after arteriotomy and stent-graft was deployed into AAA of below the renal artery to occlude the left over cavity of AAA. The stent- graft was extended and anchored to the both side wall of AAA, the blood flow enter into the arteria iliaca communis through the sten't.Results Stent-graft deployment was successfully performed in all the patients. Immediate aortography after the procedure showed no leakage in 20 patients and the type Ⅰ minor leakage in 4 patients. No stent movement or organ and both lower extremities ischemia was found at the early post operative stage in all the patients. Six months after the operation, in all the 24 patients, contrast-enhanced CT scan showed the disappearance of the aneurysm and thrombosis at the level of the stent. Conclusions EVGE is simple,minimally invasive,less complication and quick recovery after operation. Thus it becomes first choice for the treatment of AAA for the elder patients.

Gliomas are extremely aggressive brain tumors with a very poor prognosis. One of the more promising strategies for the treatment of human gliomas is targeted immunotherapy where antigens that are unique to the tumors are exploited to generate vaccines. The approach, however, is complicated by the fact that human gliomas escape immune surveillance by creating an immune suppressed microenvironment. In order to oppose the glioma imposed immune suppression, molecules and pathways involved in immune cell maturation, expansion, and migration are under intensive clinical investigation as adjuvant therapy. Toll-like receptors (TLRs) mediate many of these functions in immune cell types, and TLR agonists, thus, are currently primary candidate molecules to be used as important adjuvants in a variety of cancers. In animal models for glioma, TLR agonists have exhibited antitumor properties by facilitating antigen presentation and stimulating innate and adaptive immunity. In clinical trials, several TLR agonists have achieved survival benefit, and many more trials are recruiting or ongoing. However, a second complicating factor is that TLRs are also expressed on cancer cells where they can participate instead in a variety of tumor promoting activities including cell growth, proliferation, invasion, migration, and even stem cell maintenance. TLR agonists can, therefore, possibly play dual roles in tumor biology. Here, how TLRs and TLR agonists function in glioma biology and in anti-glioma therapies is summarized in an effort to provide a current picture of the sophisticated relationship of glioma with the immune system and the implications for immunotherapy.
Animals ; Antigens, Neoplasm ; chemistry ; immunology ; Antineoplastic Agents ; chemistry ; immunology ; therapeutic use ; Brain Neoplasms ; genetics ; immunology ; pathology ; therapy ; Chemotherapy, Adjuvant ; Clinical Trials as Topic ; Disease Models, Animal ; Gene Expression Regulation, Neoplastic ; drug effects ; immunology ; Glioma ; genetics ; immunology ; pathology ; therapy ; Humans ; Immunotherapy ; methods ; Signal Transduction ; Toll-Like Receptors ; agonists ; genetics ; immunology

Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.

Objective:To compare the efficacies of robot-assisted unilateral and manual unilateral/bilateral puncture kyphoplasty (PKP) for the treatment of osteoporotic thoracolumbar fracture (OTLF).Methods:A retrospective cohort study was conducted to analyze the clinical data of 64 OTLF patients admitted to First Affiliated Hospital of Kunming Medical University from April 2021 to May 2022. The patients included 28 males and 36 females, aged 57-88 years [(74.5±5.6)years]. Fracture segments were 12 patients from T 1-T 9, 32 from T 10-L 2, and 20 from L 3-L 5. All the patients were treated with PKP. Among them, 25 patients underwent manual unilateral puncture (manual unilateral group), 18 patients underwent manual bilateral puncture (manual bilateral group), and 21 patients underwent robot-assisted unilateral puncture (robot-assisted unilateral group). The operation time, channel establishment time, intraoperative blood loss, intraoperative fluoroscopy times, bone cement injection volume, and bone cement spatial distribution score were compared among the three groups. The visual analogue score (VAS), Oswestry disability index (ODI) and Cobb angle of kyphosis were compared among the three groups before operation, at 3 days and 3 months after operation, and at the last follow-up. The incidence of complications was compared. Results:All the patients were followed up for 6-10 months [(7.0±0.9)months]. The operation time of the manual unilateral group was (30.2±6.1)minutes, which was shorter than (37.9±8.9)minutes of the robot-assisted unilateral group and (49.0±10.2)minutes of the manual bilateral group; the operation time of the robot-assisted unilateral group was markedly shorter than that of the manual bilateral group (all P<0.05). The channel establishment time of the robot-assisted unilateral group was (4.7±1.4)minutes, which was markedly shorter than (10.4±4.4)minutes of the manual unilateral group and (21.7±6.2)minutes of the manual bilateral group (all P<0.05). The intraoperative blood loss of the robot-assisted unilateral group was (23.8±7.2)ml, which was less than (34.3±7.7)ml of the manual unilateral group and (55.9±18.7)ml of the manual bilateral group (all P<0.05). The number of intraoperative fluoroscopy of the robot-assisted unilateral group was (12.1±2.5)times, which was markedly less than (21.2±5.9)times of the manual unilateral group and (39.6±9.5)times of the manual bilateral group (all P<0.05). The channel establishment time, intraoperative blood loss and intraoperative fluoroscopy times of the manual unilateral group were markedly shorter or less than those of the manual bilateral group (all P<0.05). The bone cement injection volume and bone cement distribution score of the robot-assisted unilateral group were (4.7±1.3)ml and (7.9±1.2)points, which were not statistically different from (5.7±1.3)ml and (8.7±1.1)points of the manual bilateral group (all P>0.05), but were markedly higher than (3.0±1.3)ml and (5.1±1.8)points of the manual unilateral group (all P<0.05). There were no significant differences in VAS, ODI and Cobb angle among the three groups at 3 days, 3 months after operation and at the last follow-up (all P>0.05), but which were all lower than those before surgery (all P<0.05). There were no significant differences in VAS, ODI and Cobb angle among three groups before operation, at 3 days, 3 months after surgery and at the last follow-up (all P>0.05). The complication rate was 4.8% (1/21) of the robot-assisted unilateral group, 32.0% (8/25) of the manual unilateral group, and 33.3% (6/18) of the manual bilateral group, with no significant difference between the manual unilateral group and the manual bilateral group ( P>0.05), but both of which was markedly higher than that of the robot-assisted unilateral group ( P<0.05). Conclusion:Robot-assisted unilateral puncture and manual unilateral/bilateral puncture PKP can both achieve satisfactory results for the treatment of OTLF, but robot-assisted unilateral puncture has shorter channel establishment time, less intraoperative blood loss and intraoperative fluoroscopy times, and lower complication rate.

【Objective】 To explore the recruitment and retention strategy of blood donors by investigating the age composition of blood donors in some areas of China, so as to promote blood donation and enhance clinical blood supply. 【Methods】 Through the working platform of Practice Comparison Working Group of China’s Mainland Blood Collection and Supply Institutions, the average age and age composition of blood donors from 22 blood centers were collected, and statistical analysis was conducted after eliminating invalid data. 【Results】 The median average age of blood donors during the survey year was 30.02.The median age in 2.89% of the blood centers was lower than 25. The average age of different genders was statistically significant only in 2018(P<0.05). Fot first-time blood donors, the median constituent ratio of donors <25 and ≥25 years old was 54.53% and 44.28%, with median retention rate at 10.30% and 9.61%, respectively. The median overall participation rate of blood donors was 2.7%, with median participation rate of blood donors <25 years old at 5.1%. 【Conclusion】 The recruitment and retention of blood donor is crucial to enhance clinical blood supply. Blood donors <25 years old, with a longer period for future donation, should be the main target of blood donation recruitment. Meanwhile, the revision of upper age limit for blood donation is another important initiative to grow the blood donor pool.

Patients with acute heart failure (AHF) often experience dyspnea, and monitoring and quantifying their breathing patterns can provide reference information for disease and prognosis assessment. In this study, 39 AHF patients and 24 healthy subjects were included. Nighttime chest-abdominal respiratory signals were collected using wearable devices, and the differences in nocturnal breathing patterns between the two groups were quantitatively analyzed. Compared with the healthy group, the AHF group showed a higher mean breathing rate (BR_mean) [(21.03 ± 3.84) beat/min vs. (15.95 ± 3.08) beat/min, P < 0.001], and larger R_RSBI_cv [70.96% (54.34%-104.28)% vs. 58.48% (45.34%-65.95)%, P = 0.005], greater AB_ratio_cv [(22.52 ± 7.14)% vs. (17.10 ± 6.83)%, P = 0.004], and smaller SampEn (0.67 ± 0.37 vs. 1.01 ± 0.29, P < 0.001). Additionally, the mean inspiratory time (TI_mean) and expiration time (TE_mean) were shorter, TI_cv and TE_cv were greater. Furthermore, the LBI_cv was greater, while SD1 and SD2 on the Poincare plot were larger in the AHF group, all of which showed statistically significant differences. Logistic regression calibration revealed that the TI_mean reduction was a risk factor for AHF. The BR_ mean demonstrated the strongest ability to distinguish between the two groups, with an area under the curve (AUC) of 0.846. Parameters such as breathing period, amplitude, coordination, and nonlinear parameters effectively quantify abnormal breathing patterns in AHF patients. Specifically, the reduction in TI_mean serves as a risk factor for AHF, while the BR_mean distinguishes between the two groups. These findings have the potential to provide new information for the assessment of AHF patients.
Humans ; Heart Failure/diagnosis* ; Prognosis ; Respiration ; Wearable Electronic Devices ; Acute Disease

The significant clinical feature of bisphosphonate-related osteonecrosis of the jaw (BRONJ) is the exposure of the necrotic jaw. Other clinical manifestations include jaw pain, swelling, abscess, and skin fistula, which seriously affect the patients' life, and there is no radical cure. Thus, new methods need to be found to prevent the occurrence of BRONJ. Here, a novel nanoparticle, tFNA-KLT, was successfully synthesized by us, in which the nanoparticle tetrahedral framework nucleic acid (tFNA) was used for carrying angiogenic peptide, KLT, and then further enhanced angiogenesis. TFNA-KLT possessed the same characteristics as tFNA, such as simple synthesis, stable structure, and good biocompatibility. Meanwhile, tFNA enhanced the stability of KLT and carried more KLT to interact with endothelial cells. First, it was confirmed that tFNA-KLT had the superior angiogenic ability to tFNA and KLT both in vitro and in vivo. Then we apply tFNA-KLT to the prevention of BRONJ. The results showed that tFNA-KLT can effectively prevent the occurrence of BRONJ by accelerating angiogenesis. In summary, the prepared novel nanoparticle, tFNA-KLT, was firstly synthesized by us. It was also firstly confirmed by us that tFNA-KLT significantly enhanced angiogenesis and can effectively prevent the occurrence of BRONJ by accelerating angiogenesis, thus providing a new avenue for the prevention of BRONJ and a new choice for therapeutic angiogenesis.
Angiogenic Proteins/therapeutic use* ; Bisphosphonate-Associated Osteonecrosis of the Jaw/prevention & control* ; Endothelial Cells ; Humans ; Nanoparticles ; Nucleic Acids/therapeutic use*