Objective This study was designed to explore the influence about apoptosis of STI571 on NB4.Methods After NB4 cells were treated with STI571 in different concentration(0.5,1.0,5.0umol/L)at indicated time(24h,48h,72h)(1)morphological observation:To determine cell morphology by light microscope.(2)MTT assay:Inhibition of proliferation was measured with a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheny tetrazolium bromide(MYa3 assay.(3)Flow cytometric analysis:The expression of survivin and smac were measured on NB4 by FCM.Results Morphological observation showed characteristic apoptosis changes.MTT assay showed that STI at (0.5,1.0,5.0 μmol/L) range for 24 h,48 h and 72 h can markedly inhibit the proliferation of NB4 cells in a dose-dependent manner as well as a time-dependent manner(P＜0.05).FCM showed the expression of survivin decrease and smac increase.Conclusion STI571 In Vitro inhibits proliferation of NB4.

Objective To study the important virulence regulation genes of Brucella,and to understand their function.Methods Quantitative RT-PCR was used to quantify their relative transcription profiles under stress conditions and during macrophage cell infection.Results These genes were activated at different levels under these conditions and during cell infection,indicating their roles in pathogenesis at different srage of infection.Conclusion The transcription profiles of these genes have different effects about their functions.

Objective:To explore the clinical significance of serum nuclear factor κ B, interleukin(IL)-4, IL-10, IL-12, interferon(IFN)- γ expression in subclinical hypothyroidism with gestational diabetes mellitus.Methods:Thirty pregnant women with subclinical hypothyroidism combined with gestational diabetes mellitus in Tangshan Maternal and Child Health Hospital from January 2017 to October 2018 were retrospectively analyzed as group A. Thirty three pregnant women with subclinical hypothyroidism were selected as group B, 35 pregnant women with gestational diabetes mellitus as group C and 40 healthy pregnant women as control group.ELISA was used to detect NF-κB, IL-4, IL-10, IL-12 and IFN-γ, and the results were analyzed and compared.Results:The serum levels of NF-κB were (15.91±5.68), (13.22±5.23), (12.97±5.11), (9.74±3.85) μg/L, IL-12 were (28.91±6.84), (21.64±5.72), (22.23±5.91), (13.68±3.76) ng/L, and serum IFN-γ levels were (23.74±5.55), (18.26±4.63), (17.85±4.31), (12.69±3.85) ng/L in A, B, C and the control group respectively.There were statistically significant differences in the three indicators between groups ( F=5.118, 6.821, 7.133, all P<0.05), and group A was higher than group B, C and control, the differences were statistically significant (all P<0.05); the levels of serum IL-4 in group A, B, C and control group were (8.91±3.99), (10.84±4.47), (11.27±4.62), (13.68±5.46) ng/L, respectively.The levels of serum IL-10 were (10.91±3.86), (13.05±4.58), (12.83±4.69), (15.82±5.33) ng/L, respectively.The differences of serum IL-4 and IL-10 between groups were statistically significant ( F=5.075, 5.616, all P<0.05), and serum IL-4 and IL-10 in group A were lower than those in group B, group C and control group.The levels of serum IL-4 and IL-10 in group B and C were lower than those in control group (all P<0.05). Conclusion:The activation of NF-kB signaling pathway and its related cytokines may be the influencing factors for the development of subclinical hypothyroidism with gestational diabetes mellitus.

Objective:To investigate the clinical efficacy of posteromedial approach through anterior border of tibialis posterior tendon combined with posterolateral approach in open reduction and internal fixation of Klammer type II/III posterior pilon variant fracture.Methods:A retrospective case series study was conducted on 37 patients with posterior pilon variant fracture treated by posteromedial approach through anterior border of tibialis posterior tendon combined with posterolateral approach in Jiangsu Province People's Hospital and its Liyang branch between January 2015 and December 2019. There were 17 males and 20 females, aged 25-65 years［（49.5±9.3）years］. Klammer classification was type II in 20 patients and type III in 17 patients. The quality of fracture reduction, incision healing and postoperative complications were observed. The loss of fracture reduction or implant failure was detected during the follow-up. The fracture union time was recorded. At the final follow-up, the residual pain was assessed by the visual analogue scale（VAS）and the ankle function by the American Orthopedic Foot and Ankle Society（AOFAS）ankle-hindfoot score.Results:All patients were followed up for 12-20 months［（15.6±1.2）months］. An anatomic fracture reduction was observed in all patients, showing the incision healing by first intention. No skin necrosis, posteromedial neurovascular bundle injury, deep venous thrombosis, checkrein deformity or other complications occurred. During the follow-up, there was no reduction loss or implant failure. The fracture healing time ranged from 11 to 16 weeks［（13.2±1.4）weeks］. At the final follow-up, the VAS was decreased from 6-9 points［（8.0±0.8）points］preoperatively to 0-2 points［（1.2±0.6）points］（ P<0.01）. At the final follow-up, the AOFAS ankle-hindfoot score ranged from 80 to 100 points［（89.7±4.6）points］, which showed excellent results in 15 patients and good in 22 patients, with the excellence rate of 100%. Conclusion:The posteromedial approach through anterior border of tibialis posterior tendon combined with posterolateral approach in open reduction and internal fixation of Klammer type II/III posterior pilon variant fracture can achieve good fracture reduction, with low incidence of complications, pain relief and satisfactory ankle function recovery.

Objective:To evaluate the reliability of a region-locked 3D-printed template combined with a bi-directional matching scheme in assistance of screw placement for thoracolumbar fractures.Methods:From January 2019 to March 2023, 52 patients with thoracolumbar fracture were treated at Department of Orthopedics, The People's Hospital of Liyang. They were 29 males and 23 females, with an age of (58.2±13.3) years. They were divided into a template group and a free-hand group according to the different screw placements. In the template group of 25 cases, a region-locked 3D-printed template combined with a bi-directional matching scheme was used to assist the pedicle positioning; in the free-hand group of 27 cases, the free hand screw placement was assisted only by image data and C-arm fluoroscopy. The operation time, intraoperative fluoroscopy frequency, intraoperative blood loss, complications, and placement accuracy were compared between the 2 groups. Visual analogue scale (VAS), Oswestry disability index (ODI), and anterior height ratio of the injured vertebra were compared between preoperation, 1 week postoperation, and the final follow-up, as well as between the 2 groups.Results:There were no statistically significant differences in the preoperative general data between the 2 groups, showing comparability ( P>0.05). All patients were followed up for (11.2±4.2) months. The differences were not statistically significant between the 2 groups in intraoperative blood loss, rate of complications, VAS or ODI at preoperation, 1 week postoperation, or the final follow-up, or in anterior height ratio of the injured vertebra ( P>0.05). In the template group, the operation time [(80.1±18.5) min] was significantly longer than that in the free-hand group [(69.4±16.6) min], the intraoperative fluoroscopy frequency [2 (2, 3) times] significantly lower than that in the free-hand group [3 (3, 4) times], and the placement accuracy [98.4% (127/129)] significantly higher than that in the free-hand group [91.8% (112/122)] (all P<0.05). All patients showed significant improvements in VAS, ODI and anterior height ratio of the injured vertebra at postoperative 1 week compared with the preoperative values, and the improvements at the last follow-up were significantly larger than those at postoperative 1 week ( P<0.05). No injury to the spinal cord, nerve root or blood vessel was observed postoperatively. Conclusions:In the treatment of thoracolumbar fractures, the screw placement assisted by a region-locked 3D-printed template combined with a bi-directional matching scheme is better than free-hand screw placement in terms of improved accuracy and reduced fluoroscopy, but the former incurs longer operative exposure than the latter. There is no significant difference between the 2 methods of screw placement in clinical efficacy.

Objective:To investigate the role of serotonin reuptake transporter(SERT)and P2X3 receptor of dorsal root ganglion(DRG)in regulating visceral hypersensitivity of rats with irritable bowel syndrome(IBS)by electroacupuncture(EA). Methods:Male Sprague-Dawley and SERT-/-rats were subjected to preparing IBS visceral hypersensitivity models with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.Three weeks post-modeling,interventions including EA,intrathecal injection,and EA plus intrathecal injection were applied,respectively.Hematoxylin-eosin staining and abdominal withdrawal reflex(AWR)score were used to confirm the successful establishment of the IBS model.AWR score,whole-cell patch clamp technique,and Western blotting assay were used to evaluate the changes in visceral pain sensitivity,electrophysiological properties of DRG neurons,and the expression of DRG P2X3 receptor and SERT in IBS rats. Results:Compared to the model group,the AWR score in the EA group decreased significantly(P<0.05),the resting membrane potential(P<0.05)and the number of action potentials(P<0.05)of DRG neurons reduced,and the baseline intensity increased(P<0.05);additionally,the expression of P2X3 receptor in DRG decreased(P<0.01),and the SERT expression increased(P<0.05).Compared to the P2X3 receptor agonist group,the SERT protein expression in DRG was higher in the EA group.In SERT-/-rats,the P2X3 receptor expression in DRG increased in the EA group compared to the model group(P<0.01). Conclusion:EA modulates the electrophysiological characteristics of intestinal primary sensory neurons by regulating the expression of SERT and P2X3 receptor in DRG of IBS rats.This modulation may contribute to the mechanism by which EA alleviates peripheral sensitization of visceral pain in IBS rats.

Objective To develop a diagnostic model based on deep neural network for intelligent discrimination of thyroid function. Methods A total of 1616 patients ( 283 males, 1333 females, average age:52 years) who underwent thyroid imaging between May 2016 and June 2018 were selected. According to the clinical diagnosis, the 1616 cases included 299 normal thyroid cases, 876 hyperthyroidism cases and 441 hypothyroidism cases. Feature extraction and learning training were performed on 1000 training set sam-ples by two deep neural network models ( AlexNet;deep convolution generative adversarial networks ( DCGAN) ) using deep learning algorithm. Performance verifications were implemented on 616 test set samples. The con-sistency between the verification results of the two models and the clinical diagnosis was analyzed by Kappa test. Meanwhile, the time advantage of the intelligent diagnosis models was analyzed. Results The average diagnostic time of AlexNet model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 82.29％(79/96), 94.62％(369/390), 100％(130/130), respectively. The Kappa value between results of AlexNet model and clinical diagnosis was 0.886 ( P<0.05) . The average di-agnostic time of DCGAN model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 85.42％(82/96), 95.64％(373/390), 99.23％(129/130), respectively. The Kappa value between results of DCGAN model and clinical diagnosis was 0.904 ( P<0.05) . Conclusion The deep neural network intelligent diagnosis model can quickly determine the functional status of thyroid gland in thyroid imaging, and it has a high recognition accuracy, thus providing a new method for thyroid image review.

Objective:To establish and validate a fluorescence focus assay (FFA) for rapid titration of Japanese encephalitis virus (JEV) and to evaluate its application in the production of Japanese encephalitis vaccine.Methods:Recombinant JEV non-structural protein 1 (NS1) was expressed in a prokaryotic expression system. After purification, JEV-NS1 was used to immunize rabbits to induce polyclonal antibody. FFA was established with the polyclonal antibody to titer JEV. The accuracy of FFA was validated by comparing with plaque assay, and the specificity, precision, linearity, range and robustness of FFA were also validated. Twenty-eight batches of live-attenuated Japanese encephalitis vaccine were titrated with FFA and plaque assay to analyze the relationship between the two assays.Results:FFA established with polyclonal antibody against JEV-NS1 could be used to titrate JEV, and there was no cross reaction with other viruses (tick-borne encephalitis virus, yellow fever virus, coxsackievirus A2, coxsackievirus A4). Results of the validation tests showed that FFA met the requirement of quality control for live-attenuated Japanese encephalitis vaccine. FFA was more consistency than plaque assay.Conclusions:The established FFA could be used for virus titration in the production of live-attenuated Japanese encephalitis vaccine.

Objective To investigate the clinical application value of 3D printing technology in the treatment of complex posterior lateral tibial plateau fractures using the suprafibular approach.Methods A retrospective analysis was conducted on the clinical data of 67 patients with complex posterior lateral tibial plateau fractures treated with the suprafibular approach.Patients were divided into 3D printing group(35 cases)and conventional group(32 cases)based on whether 3D printing simulation surgery was used preoperatively.The operation time,intraoperative blood loss,and intrao-perative fluoroscopy frequency were compared between the two groups.Postoperative complications such as incision infection,popliteal vessel injury,and common peroneal nerve injury were observed.Fracture healing time was recorded during follow-up,and Rasmussen scores were evaluated at 6 months postoperatively.The Hospital for Special Surgery(HSS)knee function score was assessed during the final follow-up.Results The follow-up duration for 67 patients was 14 to 22 months.One patient in each group developed postoperative incision infection,and no complications such as poplite-al vessel injury,common peroneal nerve injury,or deep venous thrombosis of the lower extremity oc-curred.The operation time,intraoperative blood loss,and intraoperative fluoroscopy frequency were significantly lower in the 3D printing group than in the conventional group(P＜0.05).There were no significant differences in fracture healing time,Rasmussen scores at 6 months postoperatively,and HSS scores during the final follow-up between the two groups(P＞0.05).Conclusion The application of 3D printing technology in the treatment of complex posterior lateral tibial plateau fractures using the suprafibular approach can optimize surgical plans,shorten operation time,and reduce intraoperative blood loss and fluoroscopy frequency.

Objective To investigate the clinical application value of 3D printing technology in the treatment of complex posterior lateral tibial plateau fractures using the suprafibular approach.Methods A retrospective analysis was conducted on the clinical data of 67 patients with complex posterior lateral tibial plateau fractures treated with the suprafibular approach.Patients were divided into 3D printing group(35 cases)and conventional group(32 cases)based on whether 3D printing simulation surgery was used preoperatively.The operation time,intraoperative blood loss,and intrao-perative fluoroscopy frequency were compared between the two groups.Postoperative complications such as incision infection,popliteal vessel injury,and common peroneal nerve injury were observed.Fracture healing time was recorded during follow-up,and Rasmussen scores were evaluated at 6 months postoperatively.The Hospital for Special Surgery(HSS)knee function score was assessed during the final follow-up.Results The follow-up duration for 67 patients was 14 to 22 months.One patient in each group developed postoperative incision infection,and no complications such as poplite-al vessel injury,common peroneal nerve injury,or deep venous thrombosis of the lower extremity oc-curred.The operation time,intraoperative blood loss,and intraoperative fluoroscopy frequency were significantly lower in the 3D printing group than in the conventional group(P＜0.05).There were no significant differences in fracture healing time,Rasmussen scores at 6 months postoperatively,and HSS scores during the final follow-up between the two groups(P＞0.05).Conclusion The application of 3D printing technology in the treatment of complex posterior lateral tibial plateau fractures using the suprafibular approach can optimize surgical plans,shorten operation time,and reduce intraoperative blood loss and fluoroscopy frequency.