OBJECTIVE: This study aimed to explore the influence of Rce1 on invasion and migration of tongue squamous cell carcinoma cells by silencing the Rce1 gene with RNA interference. METHODS: The tongue squamous cell carcinoma Cal-27 and SCC-4 cells were cultured in vitro. The small interfering RNA (siRNA) of the Rce1 gene was designed, and the Rcel gene expression was silenced vialiposome transfection. According to the siRNA transfected by liposome, the experimental group was divided into three groups, namely, Rce1-siRNA-1, Rce1-siRNA-2, and Rce1-siRNA-3 groups. Negative control group was transfected by siCON, and the blank control group was untransfected by siRNA. The Rce1, RhoA, and K-Ras gene expression levels in each group were analyzed by real-time quantitative polymerase chain reaction. The Rce1, RhoA, K-Ras, MMP-2, and MMP-9 protein expression levels were analyzed by Western blot. The invasiveness of tongue cancer cell Cal-27 and SCC-4 were determined by Transwell invasion assay, and cell migration assay was performed by cell scratch assay. RESULTS: Real-time quantitative polymerase chain reaction and Western blot results showed that compared with the negative and blank control groups, the Rce1 gene and protein expression levels in three experimental groups decreased (P<0.05). The RhoA, K-Ras gene and protein expression levels were insignificantly different among groups (P>0.05). Meanwhile, the MMP-2 and MMP-9 expression levels decreased (P<0.05). Transwell invasion assay results showed that the total number of cells in the PET film of the experimental groups was significantly decreased compared with the control group (P<0.05). The cell scratch test showed that the cell closure time of the scratch in the interference group was significantly longer than those in the control and blank groups (P<0.05). CONCLUSIONS Silencing Rce1 in vitro can effectively downregulate its expression in tongue squamous cell carcinoma cells Cal-27 and SCC-4 and reduce the migration and invasion abilities of these cells.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Endopeptidases ; metabolism ; Humans ; Neoplasm Invasiveness ; RNA Interference ; RNA, Small Interfering ; Tongue Neoplasms ; metabolism ; therapy ; Transfection

The combination of ginkgo ketoester tablet - donepezil (GD) is a popular combination commonly used in clinic for the treatment of Alzheimer's disease. To evaluate the learning and memory improving ability of different proportions of the two drugs. We optimized the ratio of GD for treatment of dementia using a mouse model. Dementia was induced by multiple neuronal damages in mice. The experimental protocols were approved by the Animal Experimental Ethical Committee of Nanjing University of Chinese Medicine and all the procedures were strictly conducted in accordance with ethical principle of animal use and care. Morris water maze, brain hematosylin-eosin staining and the changes of the neurotransmitters and related enzymes in the plasma or brain tissues were tested to determine the effect of GD on dementia mice. The results showed that the dementia mice were significantly different from the normal group in terms of behavior, pathological sections and related indicators. Compared to the dementia mice, partial administration groups could improve learning and memory ability as well as indexes in the blood and brain tissues. Both the principal component analysis and multi-attribute comprehensive index methods were used to comprehensively evaluate the total effect of GD on anti-dementia. The results showed that the combination of two drugs at the dose of 0.5 to 1 times was in a dose-effect relationship, and the dose of 1 (the clinical equivalent) had the best treatment effect. Then based on the optimal dose, GD 1∶1 had best effect, which was consistent with the clinical use of two drugs. This provides scientific basis for more effective application of the compatibility between ketoester tablet and donepezil for modern clinic medicine.

Objective: To observe the short and long term effect of addition and subtraction therapy of Shentong Zhuyutang to myofascial pain syndrome (MPS) with stagnation of blood stasis and to investigate its mechanism of action.Method: One hundred and forty-eight eligible patients were randomly divided into control group (73 cases) and observation group (75 cases) by random number table.Patients in both groups got electroacupuncture treatment.Patients in control group additionally got Yaotong capsules,4 grains/time,3 times/day.Patients in observation group additionally got addition and subtraction therapy of Shentong Zhuyutang,1 dose/day.The treatment was continued for 6 weeks in both groups,and 16 weeks follow-up was recorded.Before treatment,and at the 1^st,2^nd,3^rd,4^th,5^th,6^th week after treatment,scores of visual analogue scale (VAS) was graded.Before and after treatment,scores of JOA,Roland-Morris disability questionnaire (RDQ),body damage index assessment scale (PⅡ scale),stagnation of blood stasis,and Pittsburgh sleep quality index (PSQI) were graded.Levels of thromboxane 2(TXB₂),6-ketone-prostaglandin F_1α(6-keto-PGF_1α) were detected,and TXB₂/6-keto-PGF_1α was calculated;in addition,the recurrence was recorded and followed up.Result: By rank sum test,the clinical efficacy in observation group was better than that in control group (Z=1.969,P<0.01).Analysis of variance of repeated measures indicated that,scores of VAS were decreased gradually at the 1^st,2^nd,3^rd,4^th,5^th,6^th week in both groups after treatment (F_control=5.801,F_observation=6.649,P<0.05).Paired t-test indicated that scores of VAS in observation group were lower than those in control group at 2^nd,3^rd,4^th,5^th,6^th week (P<0.01).Scores of three dimensions of JOA scale and total score in observation group were higher than those in control group (P<0.01).Scores of RDQ,PⅡ,PSQI,stagnation of blood stasis,whole blood viscosity (high cut and low cut),plasma viscosity,fibrinogen,TXB₂,and T/K in observation group were lower than those in control group (P<0.01),and level of 6-keto-PGF_1α was higher than that in control group (P<0.01).In addition,the recurrence rate in observation group was 12%,lower than 26.03% in control group (χ²=4.745,P<0.05).Conclusion: Based on acupuncture treatment,addition and subtraction therapy of Shentong Zhuyutang can relieve the pain caused by myofascial pain syndrome (MPS) with stagnation of blood stasis,recover lumbar activity function,reduce rate of recurrence,and ameliorate blood circulation,with anti-inflammatory and analgesia effects.

The aim of this study was to investigate whether G protein-coupled estrogen receptor (GPER) could alleviate hippocampal neuron injury under cerebral ischemia-reperfusion injury (CIRI) by acting on endoplasmic reticulum stress (ERS). The CIRI animal model was established by middle cerebral artery occlusion (MCAO). Female ovariectomized (OVX) Sprague-Dawley (SD) female rats were randomly divided into 4 groups: control, ischemia-reperfusion injury (MCAO), vehicle (MCAO+DMSO), and GPER-specific agonist G1 (MCAO+G1) groups. The neurobehavioral score was assessed by the Longa score method, the morphological changes of the neurons were observed by the Nissl staining, the cerebral infarction was detected by the TTC staining, and the neural apoptosis in the hippocampal CA1 region was detected by TUNEL staining. The distribution and expression of GRP78 (78 kDa glucose-regulated protein 78) in the hippocampal CA1 region were observed by immunofluorescent staining. The protein expression levels of GRP78, Caspase-12, CHOP and Caspase-3 were detected by Western blot, and the mRNA expression levels of GRP78, Caspase-12, and CHOP were detected by the real-time PCR. The results showed that the neurobehavioral score, cerebral infarct volume, cellular apoptosis index, as well as GRP78, Caspase-12 and CHOP protein and mRNA expression levels in the MCAO group were significantly higher than those of control group. And G1 reversed the above-mentioned changes in the MCAO+G1 group. These results suggest that the activation of GPER can decrease the apoptosis of hippocampal neurons and relieve CIRI, and its mechanism may involve the inhibition of ERS.
Animals ; Apoptosis ; Brain Ischemia ; CA1 Region, Hippocampal ; cytology ; Caspase 12 ; metabolism ; Caspase 3 ; metabolism ; Endoplasmic Reticulum Stress ; Female ; Heat-Shock Proteins ; metabolism ; Neurons ; cytology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; physiology ; Receptors, G-Protein-Coupled ; agonists ; Reperfusion Injury ; Transcription Factor CHOP ; metabolism

OBJECTIVETo observe the clinical efficacy differences between different needling methods for dry eye syndrome.

METHODSSixty patients of dry eye syndrome were randomly divided into an observation group and a control group, 30 cases (60 eyes) in each group. Shangjingming (Extra), Xiajingming (Extra), Tongziliao (GB 1), Cuanzhu (BL 2), Fengchi (GB 20), Hegu (LI 4), Sanyinjiao (SP 6), Taixi (KI 3) and Taichong (LR 3) were selected in the two groups. The control group was treated with conventional acupuncture, while the observation group was treated with guiding-acupuncture. Electroacupuncture (EA) was used at bilateral Tongziliao (GB1) and Cuanzhu (BL 2), 30 min per treatment. The treatment was given three times per week. Totally 1-month treatment (12 treatments) was given. The eye symptom score, breakup time of tear film (BUT), Schirmer Ⅰ test (SⅠT) and visual analogue scale (VAS) score were compared before and after treatment in the two groups. The clinical efficacy was compared between the two groups.

RESULTSCompared before treatment, the eye symptom score, BUT, SⅠT and VAS score were improved after treatment in the two groups (all<0.001); the improvements of eye symptom score and SⅠT in the observation group were superior to those in the control group (both<0.05). The differences of BUT and VSA score between the two groups were not significant (both>0.05). The total effective rate was 86.7% (52/60) in the observation group, which was superior to 73.3% (44/60) in the control group (<0.05). .

CONCLUSIONThe conventional EA and guiding-acupuncture combined with EA are both effective for dry eye syndrome, and the efficacy of guiding-acupuncture combined with EA is superior to that of conventional EA.