Objective To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells.Methods CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin,cisplatin or their combination.The changes in colony formation ability,apoptosis,and intracellular reactive oxygen species(ROS)levels of the treated cells were analyzed using colony formation assay and flow cytometry.Western blotting was performed to detect the changes in protein expressions in the cells.The effects of pre-treatment with NAC on proliferation,apoptosis,and PI3K/AKT signaling pathway were observed in pristimerin-and/or cisplatin-treated cells.Results Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells(P<0.05).Compared with pristimerin or cisplatin alone,their combination more strongly inhibited survival and colony formation ability of the cells,increased cell apoptosis rate and intracellular ROS levels,upregulated the protein expressions of Bax,cleaved caspase-3,and cleaved PARP,and downregulated the protein expressions of Bcl-2,Mcl-1,PARP and p-PI3K and p-AKT(P<0.05).NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin,and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment(P<0.05).Conclusion Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells,the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

Objective To investigate the effect of dihydroartemisinin(DHA)for enhancing the inhibitory effect of cisplatin(DDP)on DDP-resistant nasopharyngeal carcinoma cell line HNE1/DDP and explore the mechanism.Methods CCK-8 method was used to assess the survival rate of HNE1/DDP cells treated with DHA(0,5,10,20,40,80,and 160 μmol/L)and DDP(0,4,8,16,32,64,128 μmol/L)for 24 or 48 h,and the combination index of DHA and DDP was calculated using Compusyn software.HNE1/DDP cells treated with DHA,DDP,or their combination for 24 h were examined for cell viability,proliferation and colony formation ability using CCK-8,EdU and colony-forming assays.Flow cytometry was used to detect cell apoptosis and intracellular reactive oxygen species(ROS).The expression levels of apoptosis-related proteins cleaved PARP,cleaved caspase-9 and cleaved caspase-3 were detected by Western blotting.The effects of N-acetyl-cysteine(a ROS inhibitor)on proliferation and apoptosis of HNE1/DDP cells with combined treatment with DHA and DDP were analyzed.Results Different concentrations of DHA and DDP alone both significantly inhibited the viability of HNE1/DDP cells.The combination index of DHA(5 μmol/L)combined with DDP(8,16,32,64,128 μmol/L)were all below 1.Compared with DHA or DDP alone,their combined treatment more potently decreased the cell viability,colony-forming ability and the number of EdU-positive cells,and significantly increased the apoptotic rate,intracellular ROS level,and the expression levels of cleaved PARP,cleaved caspase-9 and cleaved caspase-3 in HNE1/DDP cells.N-acetyl-cysteine pretreatment obviously attenuated the inhibitory effect on proliferation and apoptosis-inducing effect of DHA combined with DDP in HNE1/DDP cells(P<0.01).Conclusion DHA enhances the growth-inhibitory and apoptosis-inducing effect of DDP on HNE1/DDP cells possibly by promoting accumulation of intracellular ROS.

Objective To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells.Methods CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin,cisplatin or their combination.The changes in colony formation ability,apoptosis,and intracellular reactive oxygen species(ROS)levels of the treated cells were analyzed using colony formation assay and flow cytometry.Western blotting was performed to detect the changes in protein expressions in the cells.The effects of pre-treatment with NAC on proliferation,apoptosis,and PI3K/AKT signaling pathway were observed in pristimerin-and/or cisplatin-treated cells.Results Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells(P<0.05).Compared with pristimerin or cisplatin alone,their combination more strongly inhibited survival and colony formation ability of the cells,increased cell apoptosis rate and intracellular ROS levels,upregulated the protein expressions of Bax,cleaved caspase-3,and cleaved PARP,and downregulated the protein expressions of Bcl-2,Mcl-1,PARP and p-PI3K and p-AKT(P<0.05).NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin,and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment(P<0.05).Conclusion Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells,the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

Objective To investigate the effect of dihydroartemisinin(DHA)for enhancing the inhibitory effect of cisplatin(DDP)on DDP-resistant nasopharyngeal carcinoma cell line HNE1/DDP and explore the mechanism.Methods CCK-8 method was used to assess the survival rate of HNE1/DDP cells treated with DHA(0,5,10,20,40,80,and 160 μmol/L)and DDP(0,4,8,16,32,64,128 μmol/L)for 24 or 48 h,and the combination index of DHA and DDP was calculated using Compusyn software.HNE1/DDP cells treated with DHA,DDP,or their combination for 24 h were examined for cell viability,proliferation and colony formation ability using CCK-8,EdU and colony-forming assays.Flow cytometry was used to detect cell apoptosis and intracellular reactive oxygen species(ROS).The expression levels of apoptosis-related proteins cleaved PARP,cleaved caspase-9 and cleaved caspase-3 were detected by Western blotting.The effects of N-acetyl-cysteine(a ROS inhibitor)on proliferation and apoptosis of HNE1/DDP cells with combined treatment with DHA and DDP were analyzed.Results Different concentrations of DHA and DDP alone both significantly inhibited the viability of HNE1/DDP cells.The combination index of DHA(5 μmol/L)combined with DDP(8,16,32,64,128 μmol/L)were all below 1.Compared with DHA or DDP alone,their combined treatment more potently decreased the cell viability,colony-forming ability and the number of EdU-positive cells,and significantly increased the apoptotic rate,intracellular ROS level,and the expression levels of cleaved PARP,cleaved caspase-9 and cleaved caspase-3 in HNE1/DDP cells.N-acetyl-cysteine pretreatment obviously attenuated the inhibitory effect on proliferation and apoptosis-inducing effect of DHA combined with DDP in HNE1/DDP cells(P<0.01).Conclusion DHA enhances the growth-inhibitory and apoptosis-inducing effect of DDP on HNE1/DDP cells possibly by promoting accumulation of intracellular ROS.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective: To evaluate the efficacy of Tuina (Chinese therapeutic massage) at points on abdomen and back meridians in the treatment of infantile colic.Methods: A total of 120 infants with intestinal colic were randomly divided into a control group and an observation group, with 60 cases in each group. In the control group, the parents of the infants were given soothing and health education. In addition to the intervention used in the control group, the observation group was treated with Tuina at points on abdomen and back meridians once a day for 5 consecutive days as a course of treatment. The pain scale score and clinical symptoms of the two groups were recorded before and after treatment. Results: After treatment, the total effective rate of the observation group was higher than that of the control group, and the difference was statistically significant (P<0.05). The pain scale score of the observation group was lower than that of the control group, and the difference was statistically significant (P<0.05). In the 24 h behavior diary indicators, the daily attack duration, the daily attack times, and the weekly attack days in the observation group were lower than those in the control group, and the differences were statistically significant (P<0.05). Conclusion: Tuina at points on abdomen and back meridians is effective and safe in the treatment of infantile colic.

Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve in the video of unilateral axillary approach endoscopic thyroidectomy.Methods:Videos of endoscopic thyroidectomy via unilateral axillary approach in Peking Union Medical College Hospital from Jul. 1st, 2020 to May. 1st, 2021 were collected. Videos containing the recurrent laryngeal nerve were selected, and the outline of recurrent laryngeal nerve were marked by two senior thyroid surgeons and staffs. Data were divided into training set and test set in a ratio of 5:1, and classified into high, medium and low recognition group according to difficulty of recognizing the outline of the nerve. The neuron network was based on PSPNet combined with Resnet50. All data were analyzed by R (ver. 4.0.2) .Results:A total of 38 videos including 35,501 frames of pictures were included in this study. 29, 704 frames of 32 videos were in our training set and 5797 frames of 6 videos were in the test set. When the intersection over union (IOU) threshold is 0.1, the sensitivity and precision is 100.0%/92.1%, 95.8%/80.2% and 81.0%/80.6% in high, medium and low recognition group respectively. When the IOU threshold is 0.5, the sensitivity and precision is 92.6%/85.3%, 71.7%/60.5% and 38.1%/37.9% in high, medium and low recognition group respectively, indicating that neuron network could located the outline of recurrent laryngeal nerve in high and medium recognition group. False negatives were often due to small targets and unclear boundaries.Conclusion:Recurrent laryngeal nerve recognition based on deep learning is feasible and has potential application value in endoscopic thyroidectomy, which may help surgeons reduce the risk of accidental injury of recurrent laryngeal nerve and improve the safety of thyroidectomy.

Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve (RLN) in videos of endoscopic thyroidectomy (ETE) via breast approach.Methods:Videos of ETE via breast approach in Peking Union Medical College Hospital from Feb. 2020 to Aug. 2021 were collected. Videos containing RLN were selected, and the outline of RLN was marked by two thyroid surgeons. Then data were divided into a training set and a test set in a ratio of 5:1 and classified into the high and low difficulty group according to a senior thyroid surgeon’s opinion. Those pictures were input to D-LinkNet model. Precision, sensitivity and mean dice index was calculated.Results:A total of 46 videos including 153, 520 frames of pictures were included in this study. 131,039 frames of 39 videos were in the training set and 22,481 frames of 7 videos were in the test set. When the intersection over union threshold was 0.1, the sensitivity and precision was 92.9%/72.8% and 47.6%/54.9% in high and low recognition group, respectively. When the intersection over union threshold was 0.5, the sensitivity and precision turned to 85.8%/67.2% and 37.6%/43.5% in high and low difficulty group, respectively. Mean Dice index was 0.781 and 0.663 in high and low difficulty group, respectively.Conclusions:RLN recognition based on deep learning is feasible and has potential application value in ETE, which may help surgeons reduce the risk of accidental injury of RLN and improve the safety of thyroidectomy.

OBJECTIVE: To investigate the effect of down-regulation of miR-205-5p on 3-bromopyruvate-induced apoptosis in human nasopharyngeal carcinoma CNE2Z cells. METHODS: Nasopharyngeal carcinoma CNE2Z cells were transfected with miR- 205-5p-mimic or miR-205-5p-inhibitor, treated with 80 μmol/L 3-bromopyruvate alone, or exposed to both of the treatments. The proliferation of the treated cells was examined with MTT assay, and early apoptosis of the cells was detected using a mitochondrial membrane potential detection kit (JC-1). DAPI fluorescence staining was used to detect morphological changes of the cell nuclei and late cell apoptosis; Annexin V-FITC/PI double staining was employed to detect the cell apoptosis rate. Western blotting was used to detect the expressions of Bcl-2, Bax, Mcl-1 and Bak proteins. RESULTS: Exposure to 3-bromopyruvate significantly inhibited the proliferation of CNE2Z cells, and increasing the drug concentration and extending the treatment time produced stronger inhibitory effects. Treatment with 80 μmol/L 3-bromopyruvate for 24, 48 and 72 h resulted in inhibition rates of (45.7±1.21)%, (64.4±2.02)% and (78.3±1.55)% in non-transfected CNE2Z cells, respectively; the inhibition rates were (27.7±1.04)%, (34.8±2.10)% and (44.3±1.57)% in the cells transfected with miR-205-5p-mimic, and were (80.5 ± 0.94)%, (87.9 ± 0.50)% and (93.8 ± 1.16)% in cells transfected with miR-205-5p-inhibitor, respectively. The results of mitochondrial membrane potential detection showed that the relative proportion of red and green fluorescence decreased significantly in miR-205-5p-inhibitor-transfected cells with 3-bromopyruvate treatment. Combined treatment of the cells with 3-bromopyruvate and miR-205-5p-inhibitor transfection obviously increased nuclear fragmentation and nuclear pyknosis and significantly increased cell apoptotic rate as compared with the two treatments alone ( < 0.01), causing also decreased expressions of Bcl-2 and Mcl-1 proteins and increased expressions of Bax and Bak proteins. CONCLUSIONS Inhibition of miR-205-5p enhances the proapototic effect of 3-bromopyruvate in CNE2Z cells possibly in relation to the down-regulation of Mcl-1 and Bcl-2 and the up-regulation of Bak and Bax proteins.

OBJECTIVETo observe the differences in the clinical therapeutic effects on idiopathic tinnitus between the combined therapy with acupuncture and the modified formula of and and the simple administration of the modified formula of and .

METHODSA total of 60 patients were randomized into a combined treatment group and an herbal medicine group, 30 cases in each one and 2 cases dropped out in the herbal medicine group. In the combined treatment group, acupuncture was given at the local acupoints for tinnitus, such as Yifeng (TE 17), penetrating needling technique from Ermen (TE 21) to Tinggong (SI 19) and Tinghui (GB 2) combined with the scalp acupuncture [the vertigo-auditory area, Baihui (GV 20)] and the oral administration of the modified formula of and . In the herbal medicine group, the oral administration of the modified formula of and was only applied. The treatment was given once a day in each group, 5 times a week for acupuncture and 7 times a week for the oral administration of herbal medicine. Totally, the treatment for 6 weeks was required in the two groups. Before and after treatment, the tinnitus severity score (TSS) and the score of tinnitus handicap inventory (THI) were observed and the clinical therapeutic effects were compared between the two groups.

RESULTSAfter treatment, the TSS and THI scores reduced as compared with those before treatment in the two groups (all <0.05). The scores in the combined treatment group were lower than those in the herbal medicine group (both <0.05). The total effective rate in the combined treatment group was 93.3% (28/30), better than 67.9% (19/28) in the herbal medicine group (<0.05).

CONCLUSIONThe combined treatment with scalp acupuncture, acupuncture around the ear and the modified formula of and achieve the superior therapeutic effects on idiopathic tinnitus as compared with the simple oral administration of the modified formula of and .

Acupuncture Points ; Acupuncture Therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Phytotherapy ; Tinnitus ; therapy ; Treatment Outcome