This paper presents an automatic acquisition and analytic procedure of acupuncture manipulation based on optical navigation, aiming at solving the shortcomings of existing acquisition methods of acupuncture manipulation. An acquisition holder installed at the handle tail of filiform needle was designed to display the movement trajectory of the needle during acupuncture delivery by collecting the movement trajectory of holder. The 3-month old male Bama miniature pig was selected as the experimental subject, and 6 points, "Bojian" "Qiangfeng" "Housanli" "Xiaokua" "Huiyang" (BL35) and "Baihui" (GV20), were selected during acupuncture manipulation. The optical navigation system was used to collect the real-time data, and these data were per-processed and analyzed using mean filtering and Fourier transform. The acupuncture procedure was divided into 3 stages, inserting, lifting-thrusting, and twisting. The results showed that the accuracy was 96.3% at lifting-thrusting stage, and that was 100.0% at twisting stage. The decomposition effect of the entire procedure was satisfactory. This study provides a new approach to the quantitative analysis of acupuncture manipulation. In the future, it needs to further optimize the algorithm and expand the sample size so as to improve the accuracy of this analytic technique.
Acupuncture Therapy/methods* ; Male ; Animals ; Swine ; Acupuncture Points ; Humans ; Swine, Miniature ; Needles

Objective: To investigate the therapeutic effect of bone marrow mesenchymal stem cells ( BMSCs) mod- ified by bone morphogenetic protein ( BMP) -2 gene combined with nano-hydroxyapatite ( nHA) / polyamide 66 ( PA66) on femoral nonunion in rats． Methods : Rat BMSCs were isolated and divided into the stent + BMSCs group，stent + BMSCs + rhBMP-2 group，and stent + BMSCs + Ad-BMP-2 group; human recombinant BMP-2 ( rh- BMP-2) or adenovirus-infected BMP-2 ( Ad-BMP-2) vector was transfected into BMSCs and loaded onto nHA / PA66 stent materials．Flow cytometry was used to detect that BMSCs expressed specific surface markers．Cell pro- liferation was detected by MTT assay，related bioactive factors［platelet-derived growth factor ( PDGF) ，transfor- ming growth factor-β ( TGF-β) ，vascular endothelial growth factor ( VEGF) ，fibroblast growth factor ( FGF) ，os- teocalcin ( OCN) ，osteonectin ( ON) ］were detected by ELISA，alkaline phosphatase ( ALP) activity was detec- ted，and cell growth and adhesion were observed by scanning electron microscopy ( SEM) ．In the atrophic nonun- ion model of SD rats，the stent + BMSCs + rhBMP-2 or stent + BMSCs + Ad-BMP-2 complex was implanted into the defect area，which was divided into the rhBMP-2 group and the Ad-BMP-2 group，and the therapeutic effect was e- valuated by X-ray and MicroCT． Results: The surface of the nHA / PA66 stent was smooth and porous，and BMSCs adhered well．Flow cytometry showed high expression of CD29 and CD90 and low expression of CD45 and CD34 in BMSCs．MTT showed that the cells proliferated rapidly after 72 h．Compared with the stent + BMSCs group and the stent + BMSCs + rhBMP-2 group，the ALP activity，the expressions of PDGF，TGF-β , VEGF，FGF，OCN，and ON in the stent + BMSCs + Ad-BMP-2 group were significantly up-regulated ( P＜0. 05) ．12 weeks after surgery， the stent complex in the Ad-BMP-2 group of rats was completely wrapped by newly formed cortical bone，and the surface was smooth and intact．X-ray showed that the complex in the Ad-BMP-2 group was completely wrapped by newly formed cortical bone，and the recovery degree was better than that in the rhBMP-2 group．Micro-CT results showed that compared with the rhBMP-2 group，the bone volume fraction，trabecular thickness，trabecular num- ber，bone mineral density，and bone mineral content in the Ad-BMP-2 group all increased 12 weeks after surgery ( P＜0. 05) ，and the trabecular separation level decreased ( P＜0. 05) ． Conclusion Ad-BMP-2-transfected BM- SCs combined with nHA / PA66 stent material can express bioactivity more effectively，provide a continuous and good microenvironment for the proliferation and differentiation of BMSCs，which is beneficial to promoting local os- teogenic activity and bone defect repair．

OBJECTIVE: To evaluate the performance of a newly developed intelligent acupuncture robot that simulates human manual acupuncture techniques through an animal experiment using miniature pigs. METHODS: Two 3-month-old Bama miniature pigs were selected. One pig was used for the manual needling techniques of the practitioner, and the other pig was used for the intelligent acupuncture robot. The acupoints selected were "Qiangfeng" "Bojian" "Xiaokua" "Huiyang" (BL 35) and "Baihui" (GV 20), with a straight insertion depth of 25-35 mm. The manipulation techniques, including lifting-thrusting and twisting at a frequency of 90 times/min, were applied for 1 min, with 30 s each for lifting-thrusting (10 mm amplitude) and twisting (180° angle). The practitioner's needling techniques were captured using an optical motion capture system, and the collected data were input into the intelligent acupuncture robot for replication on the second miniature pig. A complete needling session of five acupoints was considered one round, and three rounds were performed per session, every other day, for a total of five sessions, completing 15 rounds. The Fréchet distance method was used to compare the robot's replicated needling data with the practitioner's data, and curve fitting analysis was conducted to evaluate the robot's performance and precision. RESULTS: The intelligent acupuncture robot achieved a total completion rate of 96.00% for 75 needling operations, with a non-completion rate of 4.00%. The robot's replication of the practitioner's needling techniques showed a good fit, with many characteristic points overlapping. The average Fréchet distance was 18.67. The Fréchet distance for the lifting-thrusting and twisting techniques at Baihui (GV 20) and "Huiyang" (BL 35) acupoints was smaller than that at "Xiaokua" and "Qiangfeng" points (P<0.01). The accuracy of the lifting-thrusting technique at "Xiaokua" and "Qiangfeng" acupoints was lower than the twisting technique (P<0.01). CONCLUSION The intelligent acupuncture robot is able to replicate the pre-recorded manual needling parameters with a high degree of accuracy.
Animals ; Swine ; Swine, Miniature ; Acupuncture Therapy/methods* ; Robotics/instrumentation* ; Humans ; Acupuncture Points ; Female ; Male

Objective To investigate the efficiency of biological function of AT rich interaction domain 1A (ARID1A) gene silenced by small interfering RNA (siRNA) on ovarian clear cell carcinoma ES2 cell line. Methods (1) The three pairs ARID1A gene siRNA interference fragments siN1 (ARID1A-705), siN2 (ARID1A-1513), siN3 (ARID1A-2282) and one pair negative control were respectively designed, and transfected into ES2 cells by RNA interference max reagent transiently. Reverse transcription (RT)-PCR and western blot methods were used to detect the expression of ARID1A mRNA and protein in ES2 cells transfected with interference fragments respectively. So as to select the best silencing effect of siRNA interference fragment(that was siN3),and then was used in the following experiment. (2) The following experiment were divided into three groups, namely siN3 transfection group, negative control group and blank control group. The proliferative activity of three groups of cells after transient transfection ( 6, 24, 48, 72, 96 hours) was assessed by cell counting kit-8 (CCK-8) assay and expressed as absorbance (A) value; the apoptosis rate of three groups of cells transfected transiently with interference fragment was measured by flow cytometry with annexin V/propidium iodide (PI) staining;the ability of cellular invasion of three groups of cells transfected transiently with interference fragment was tested by transwell experiment;the expression of nuclear factor-kappa B (NF-κB), membrane type-1 matrix metalloproteinase (MT1-MMP) and matrix metalloproteinase-2 (MMP2) protein in ES2 cells transfected transiently with interference fragment was measured by western blot. Results (1) The RT-PCR results showed that the ARID1A mRNA relative expression levels in ES2 cells after transfected transiently with siN1, siN2 and siN3 were 0.007 8±0.005 7, 0.006 8±0.000 3 and 0.002 8±0.000 3 respectively. They were all apparently lower than that in the negative control group (0.034 6 ± 0.001 3;all P<0.01). The western blot results showed that the expression levels of ARID1A protein were 0.439 4±0.000 7, 0.424 4±0.005 0 and 0.386 0±0.005 8 respectively. They were also lower than that in the negative control group (0.732 4 ± 0.030 3; all P<0.01). The siN3 with the highest transfection efficiency was selected to use in the following experiment. (2) The CCK-8 method showed that the proliferative activity of siN3 transfection group cells after transfected transiently at 6 hours was not statistically significant difference compared with those in negative control group and blank control group (0.506 ± 0.010, 0.491 ± 0.006, 0.498 ± 0.009, respectively; all P>0.05). However, the proliferative activity of siN3 transfection group cells after transfected transiently at 24, 48, 72, 96 hours were higher than those in negative control group and blank control group (all P<0.01). The flow cytometry results showed that the apoptosis rate of siN3 transfection group cells was (20.0±3.9)%, which was significantly lower than those in negative control group and blank control group [(31.5 ± 5.0)%, (34.0 ± 4.2)%, respectively;all P<0.05]. The transwell experiment showed that the penetrated cell counts of siN3 transfection group was 60.4±2.9, which was apparently higher than those in negative control group and blank control group (54.2 ± 3.5, 52.1 ± 3.8, respectively; all P<0.01). Western blot experiment showed that the relative expression levels of NF-κB, MT1-MMP and MMP2 protein in siN3 transfection group were respectively 1.85 ± 0.16, 0.37 ± 0.08, 1.38 ± 0.11, which were apparently higher than those in negative control group (0.93±0.11, 0.17±0.05, 0.86±0.06;all P<0.05) and blank control group (0.94 ± 0.04, 0.15 ± 0.08, 0.85 ± 0.10, respectively; all P<0.01). Conclusions It would be to promote the cell doubling time, reduce cell apoptosis and increase the invasive capability in ES2 cells that ARID1A expression was down-regulating by ARID1A mRNA interference. The invasion mechanism may be related to the activation of NF-κB signal transduction pathway, up-regulation of MT1-MMP expression and then promoting the invasion of tumor cells via the up-regulation of MMP2 expression.

Objective To study the antitumor effect of new photodynamic therapy (PDT) applying TMPyP4 combined with nucleolin silence on cervical cancer SiHa cells in vitro, and to explore an available combination treatment project for cervical cancer.Methods The SiHa Cells were divided into blank control group, RNAi group,PDT group and PDT-RNAi group.The proliferation activities of SiHa cells were assessed by Cell Counting Kit-8 (CCK8)assay.The apoptotic rates were measured by flow cytometry (FCM)with Annexin Ⅴ/PI staining. The invasiveness abilities were assessed by Transwell assay. Results Compared with blank control group, the inhibitory rates of growth of SiHa cells in RNAi group, PDT group and PDT-RNAi group were increased significantly (P<0.01);the inhibitory rates of growth of SiHa cells in PDT-RNAi group were higher than those in PDT group and RNAi group (P<0.05 ). The Q value was 1.27. Compared with blank control group, the apoptotic rates of SiHa cells in experiment groups were increased (P<0.05 ), and the late apoptotic rate in PDT-RNAi group was also increased;there were significant differences of the apoptotic rates between PDT-RNAi group and RNAi group, PDT group (P<0.05 ). Compared with blank control group, the cell invasiveness abilities of SiHa cells in experiment groups were decreased;there were significant differences of the invasiveness abilities of SiHa cells between PDT group and RNAi group (P<0.05).Conclusion New PDT shows a strong photodynamic effect on the SiHa cells,which can inhibit the proliferation and invasiveness and induce the apoptosis of SiHa cells invitro;nucleolin silence shows a good synergy effect to PDT.