Objective To investigate the effect of silencing Notch1 gene by RNA interference on the proliferation,apoptosis and Akt/mTOR signaling pathway in clear renal cell carcinoma.Methods The optimal segment targeting Notch1 gene was designed and transfected into 786-O cells by Lipofectamine TM2000.The Notch1 mRNA and protein were detected by RT-PCR and Western blot.The proliferation rate of 786-O cells was evaluated by MTT and the variation of apoptosis was measured by TUNEL.The protein expression level of apoptosis-related protein Bcl-2,caspase-3,caspase-9,and signaling pathway protein Akt,p-Akt,p-mTOR,p-P70S6K were detected by Western blott.Results Notchl mRNA and protein was markedly suppressed by the siRNA targeting Notch1.Treated with 0,40,60,80,100 and 120 nmol/L of Notch1 siRNA for 24 hours,cell proliferation rates were (98.51 ± 1.33) ％,(87.34 ± 2.26) ％,(64.72 ± 3.24)％,(57.68 ±3.32)％,(31.91 ± 1.85)％ and (19.27 ±2.73)％,and the difference was statistically significant (P ＜ 0.01).Treated with 0,40,80,and 120 nmol/L of Notchl siRNA for 24 hours,apoptosis rates were (7.6 ± 3.8) ％,(21.5 ± 4.8) ％,(32.3 ± 3.5) ％,and (46.3 ± 4.7％),the difference was statistically significant (P ＜ 0.05).Decreased expression of Akt signaling pathway proteins p-Akt,p-mTOR,p-70S6K and apoptosis-related protein Bcl-2,procaspase-3 was detected,but no change in the total protein of Akt.Conclusions Depletion of Notch1 gene could inhibit cell growth and induce apoptosis in 786-O cell line.It inhibits Akt/mTOR signaling pathway by dephosphorylated.

Objective:To investigate the effect of MT01 on the differentiation of osteoblasts under infected condition through determining the expression level of collagen Ⅰ (Col Ⅰ )mRNA in MG63 cells treated with Porphyromonas gingivalis (Pg).Methods:The cultured MG63 cells were divided into blank control,MT01,Pg, and MT01+Pg groups.MT01 at a concentration of 1 mg·L-1 was added into the MG63 cells,and the cells were incubated for 3 h.The cells treated with PBS (1 mg·L-1 )were used as control group.Then Pg (MOI=100∶1) was added.Real-time PCR was used to detect the expression levels of ColⅠ mRNA in MG63 cells at 2,4,6,8, 12 and 24 h after incubation.Results:Compared with blank control group,the levles of ColⅠ mRNA in the MG63 cells in MT01 and MT01 + Pg groups had no significant changes at 2 and 4 h (P > 0.05);the Col Ⅰ mRNA expression levels in MT01 group at 8,12 and 24 h were increased (P < 0.05 or P < 0.01).Compared with Pg group,the expression levels of ColⅠ mRNA in MT01+ Pg at 2 and 4 h were decreased,but the expression levels of ColⅠ mRNA were increased at 6,8,12 and 24 h (P <0.05 or P <0.01).Conclusion:MT01 can up-regulate the expression level of Col Ⅰ mRNA in the infected MG63 cells; MT01 could promot the differentiation of osteoblasts under infected condition.

Objective To investigate the effect of methylene blue (MB) on oxygen metabolism in patients with septic shock. MethodsForty ASA Ⅱ or Ⅲ patients with septic shock aged 38-64 yr weighing 48-75 kg undergoing emergency surgery were randomly divided into 2 groups ( n = 20 each): group Ⅰ norepinephrine (group NE) and group Ⅱ MB. The patients were unpremedicated. Anesthesia was induced with midazolam, etomidate,sufentanil and vecuronium and maintained with inhalation of 0.5％-1.5％ sevoflurane and intermittent iv boluses of sufentanil and vecuronium. The patients were mechanically ventilated after tracheal intubation. PETCO2 was maintained at 35-45 mm Hg. During operation MB was infused at 0.5-1.0 mg·kg-1·h-1 in group MB and NE at 0.5-2.0 μg· kg-1 · min-1 in group NE respectively to maintain hemodynamic stability. Radial artery was cannulated and connected with Vigileo cardiac output monitor. Right internal jugular vein was cannulated for CVP monitoring.HR, SvO2, MAP, CVP, SV and CI were continuously monitored. Arterial and central venous blood samples were collected simultaneously before induction of anesthesia (T0, baseline), immediately before (T1) and at 30, 60 and 90 min after skin incision (T2-4) and at the end of operation (T5) . Blood gas analysis was performed. O2 consumption index (VO2I), O2 delivery index (DO2I) and O2 extraction rate (ERO2) were calculated. Blood lactate concentration was measured. Results MAP, HR, CVP, SVRI, DO2I, VO2I and ERO2 were significantly higher,while CI and blood lactate concentration lower during operation (T2-5) in group MB than in group NE. MAP, HR,CVP, SVRI, VO2I, DO2I, and ERO2 were significantly higher, while CI and blood lactate concentration were lower during operation (T2-5) as compared with the baseline values at T0 in group MB. In group NE there were no significant change in MAP, HR, CVP and DO2I during operation (T2-5 ) as compared with the baseline at T0. ConclusionIntravenous infusion of MB at 0.5-1.0 mg·kg-1·h-1 during operation may improve hemodynamics and oxygen metabolism in patients with septic shock.

OBJECTIVEThis aimed to investigate the effect of specific sequence oligodeoxynucleotide MT01 on the biological properties of osteoblasts invaded by Porphyromonas gingivalis (P. gingivalis ) by evaluating proliferation, cell cycle, and apoptosis.

METHODSMG63 osteoblasts were recovered and incubated with MT01, CpG ODN, metronidazole (MNZ), and gentamicin (GEN) for 3 h. P. gingivalis (the multiplicity of infection was 100:1) was added subsequently and cocultured for another 24 and 48 h. Cells with PBS comprised the blank group, whereas cells with P. gingivalis comprised the negative controls. Six experimental groups were established: PBS group, P. gingivalis group, MT01+P. gingivalis group, CpG ODN+ P. gingivalis group, MNZ+P. gingivalis group, and GEN+P. gingivalis group. The proliferative ability was measured by methyl thiazolyl tetrazolium assay, and the percentages of apoptosis and cell cycle were examined by flow cytometry.

RESULTSCompared with the blank group, proliferation increased significantly in the MT01+P. gingivalis group (P < 0.05). The ratio of cells was lower at the G₁ phase and higher at the S phase in the MT01+P. gingivalis group compared with the results in the P. gingivalis group (P < 0.05). Early cell apoptosis in the MT01+P. gingivalis group was significantly lower than that in the P. gingivalis group (P < 0.05).

CONCLUSIONMT01 can promote the proliferation, reduce the ratio of the G₁phase, increase the ratio of the S phase, and inhibit the early apoptosis of osteoblasts invaded by P. gingivalis.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Division ; Cell Proliferation ; drug effects ; Flow Cytometry ; Gentamicins ; pharmacology ; Humans ; Metronidazole ; pharmacology ; Oligodeoxyribonucleotides ; pharmacology ; Osteoblasts ; cytology ; drug effects ; Porphyromonas gingivalis ; pathogenicity

Objective:To retrieve, evaluate and integrate the relevant evidence of prevention of lower limb ischemic complications in venous-arterial extracorporeal membrane oxygenation (VA-ECMO) patients, and provide reference for the development of scientific and complete prevention and management of lower limb ischemic complications.Methods:According to the evidence-based methodology, clinical decisions, guidelines, expert consensus, evidence summary, systematic review, randomized controlled trials and experimental studies related to lower limb ischemia complications in VA-ECMO patients were searched from CNKI, Wanfang, PubMed, Cochrane Library and other domestic and foreign databases as well as relevant professional websites. The literature search period was from the establishment of the database to August 2023. Two researchers independently evaluated the literature quality, and then extracted and summarized the evidence according to the theme.Results:A total of 27 004 articles were obtained in the preliminary search, and 11 articles were finally included after screening, including 1 guide, 2 expert consensus, 2 systematic reviews and 6 original studies. Through literature reading, evidence extraction and classification, and expert group meetings, a total of 24 best evidences were concluded in four dimensions, including team training and management, VA-ECMO pre-computer evaluation, VA-ECMO catheter selection, and the monitoring and management of lower limb ischemia.Conclusions:This study summarized the best evidence for the prevention of lower limb ischemia complications associated with VA-ECMO patients, and can provide reference for healthcare providers in clinical practice. In order to ensure the safety of VA-ECMO treatment and reduce the incidence of related complications, healthcare professionals should carefully select and apply evidence according to the clinical context and patients′ wishes.

Traditional Chinese Medicine (TCM) non-drug therapy for gastroesophageal reflux mainly includes appropriate TCM techniques, such as conventional acupuncture, moxibustion, fire needle, etc. Emotional therapy, such as regulating emotion, transforming the patient's spirit and change the state of qi, the five-element music therapy, etc; exercise therapy, such as Baduanjin and Zhanzhuang, can be used alone, or in combination, or in combination with oral administration of Chinese materia medica. By reducing the probability of esophageal sphincter relaxation, inhibiting gastric acid secretion, improving esophageal motility disorder, reducing visceral hypersensitivity, immune regulation and other effects, it can alleviate the symptoms of acid reflux, heartburn and pharyngeal discomfort, and help to improve the patients' anxiety, depression and other negative emotions. It has the characteristics of simplicity and fewer adverse reactions, at the same time, according to the patient's condition and compliance to choose the appropriate therapy. The mechanism of TCM non-drug therapy in the treatment of this disease needs to be further explored, so as to better guide clinical popularization and application.

The ultrasonic manifestations of benign and malignant breast imaging-reporting and data system(BI-RADS)4 lesions overlap in some degrees,is able to result in unnecessary biopsy or untimely therapy.Accurate classifying the nature of BI-RADS 4 breast lesions can provide reliable references for clinical decision-making.The progresses of application of new ultrasonic technologies,including automated breast volume scanner,superb micro-vascular imaging,elastography,contrast-enhanced ultrasound and artificial intelligence for assisting diagnosis of BI-RADS 4 lesions were reviewed in this article.

Objective:To study the changes in the sphincter of Oddi pressure in rabbits after bilateral vagus nerve trunk severance and the therapeutic effect of Qingre Lidan Decoction, to provide a new way for the treatment of sphincter of Oddi dysfunction.Methods:Twenty-four 1.5~2.0 kg New Zealand Large White rabbits of either gender were randomly selected and divided into the control group, the model group, and the treatment group. In the control group, only pyloroplasty was performed; in the model group and the treatment group, pyloroplasty plus bilateral vagus nerve trunk dissection at the level of the diaphragm were performed, and in the treatment group, one month of gavage treatment with Qingre Lidan Decoction was carried out 7 days after the operation. Multi-channel bio-signal acquisition system was used to record the pressure changes of the sphincter of Oddi in rabbits of each group; ELISA was used to detect the changes of inflammatory factors IL-6 and tumor necrosis factor TNF-α in the serum of rabbits.Results:The pressure of the sphincter of Oddi in the model group was significantly higher than those in the control group [low-pressure area: (51.95±0.35) mmHg (1 mmHg=0.133 kPa) vs (21.60±1.13) mmHg ( P<0.05) ; High pressure area: (60.75±0.49) mmHg vs (20.70±0.85) mmHg ( P<0.05)], the pressure of sphincter of Oddi in the treatment group of Qingre Lidan Decoction was lower than that of the model group [low-pressure area: (22.70±1.13) mmHg vs (51.95±0.35) mmHg ( P<0.05); high-pressure area: (32.15±0.49) mmHg vs (60.75±0.49) mmHg ( P<0.05)]. Serum IL-6 and TNF-αwere significantly elevated in the model group compared to the control group; IL-6 and TNF-α levels were decreased in the treatment group compared to the model group, and the differences were statistically significant ( P<0.05). Conclusion:Bilateral vagus nerve trunk severance leads to Oddi sphincter dysfunction, and treatment with Qingre Lidan Decoction could improve Oddi sphincter dysfunction.

Zedoary turmeric oil， volatile oil extracted from zedoary turmeric， composed mainly of monoterpenes （including α-pinene， β-pinene， etc.） and sesquiterpenes （including β-elemene， zedoary alcohol， zedoary ketone， etc.）， and has been used in clinical practice to treat various malignant tumors such as ovarian cancer， cervical carcinoma， colorectal cancer， lung cancer and liver cancer. Zedoary turmeric oil regulates vascular endothelial growth factor and nuclear factors- κB， signal transducers and activator of transcription 3 signaling pathways to play a role in inhibiting tumor angiogenesis， inhibiting tumor cell proliferation， inducing tumor cell apoptosis， and blocking cell cycle. However， due to its insolubility in water and poor stability， its clinical application is limited； the application of new formulations and technologies such as liposomes， microspheres， and nanoemulsion improves the solubility and stability of zedoary turmeric oil. This paper summarizes recent research progress on the chemical composition， anti-tumor effects， and formulations of zedoary turmeric oil， both domestically and internationally， providing a reference for further expanding the clinical application and formulation development of zedoary turmeric oil in the anti-tumor field.

Zedoary turmeric oil， volatile oil extracted from zedoary turmeric， composed mainly of monoterpenes （including α-pinene， β-pinene， etc.） and sesquiterpenes （including β-elemene， zedoary alcohol， zedoary ketone， etc.）， and has been used in clinical practice to treat various malignant tumors such as ovarian cancer， cervical carcinoma， colorectal cancer， lung cancer and liver cancer. Zedoary turmeric oil regulates vascular endothelial growth factor and nuclear factors- κB， signal transducers and activator of transcription 3 signaling pathways to play a role in inhibiting tumor angiogenesis， inhibiting tumor cell proliferation， inducing tumor cell apoptosis， and blocking cell cycle. However， due to its insolubility in water and poor stability， its clinical application is limited； the application of new formulations and technologies such as liposomes， microspheres， and nanoemulsion improves the solubility and stability of zedoary turmeric oil. This paper summarizes recent research progress on the chemical composition， anti-tumor effects， and formulations of zedoary turmeric oil， both domestically and internationally， providing a reference for further expanding the clinical application and formulation development of zedoary turmeric oil in the anti-tumor field.