Objective To establish nasopharyngeal carcinoma cell line (CNE1) with eIF1 gene stable over-expression and to study its effects on the proliferation and migration of nasopharyngeal carcinoma cells.Methods EIF1 over-expression vector was constructed by adopting the pEGFPC1 eukaryotic expression system for transfecting nasopharyngeal carcinoma CNE1 cells.Thus the stably transfected EIF1-elF1 and its control cells were obtained.The over-expression situation of eIF1 in these cells was verified by real time fluorescence quantitative PCR(qPCR) and Western blot.The proliferation and migration activity of CNE1-eIF1 cells were tested by adopting the cell proliferation and migration tests.Results The enzyme digestion electrophoresis identification and sequencing showed that the pEGFPC1-eIF1 eukaryotic expression vector was successfully constructed.After mRNA and protein expression identification,compared with the reloading plasmid transfection group,the eIF1 gene mRNA and protein expression levels in nasopharyngeal carcinoma cell line CNE1 stably over-expressing eIF1 were up-regulated by 2.85 folds and 2.58 folds respectively (P＜ 0.05),while its proliferation and migration activities were down-regulated by 55 ％ and 36 ％ respective (P＜ 0.05).Conclusion The nasopharyngeal carcinoma cell line over-expressing elF1 is successfully constructed,the eIF 1 over-expression could significantly down-regulate the proliferation and migration activities of nasopharyngeal carcinoma cells,suggesting that eIF1 has potential anti-tumor effect.

Objective:To explore the prescription and medication law of Traditional Chinese Medicine (TCM) compounds in the treatment of vascular dementia (VD) based on patent database.Methods:TCM compounds with patents about VD were retrieved from Chinese patent announcement website of the State Intellectual Property Office and CNKI. The retrieval time was from the establishment to the databases to 31 st, March 2022. The frequency, clusteringand association analysis were carried out with the help of TCM inheritance auxiliary platform (V2.5). The medication law was analyzed. Results:154 TCM compound patents for the treatment of vascular dementia were screened, involving 227 kinds of Chinese materia medica. Among them, Acori Tatarinowii Rhizoma (44 times, 28.57%) was used more frequently, and the common medicinal pair was Salviea Miltiorrhizae Radix et Rhizoma- Acori Tatarinowii Rhizoma (17 times, 11.03%). The medicinal property was mainly warm, the taste was mainly sweet, and the meridian was mainly liver meridian. Those with high confidence based on association rules were " Corni Fructus -Acori Tatarinowii Rhizoma" (0.90), " Corni Fructus -Rehmannize Radix et Praeparata" (0.90). Based on the complex network, it was concluded that the core drugs were 14 groups such as " Rehmannize Radix et Praeparata- Cistanches Herba- Corni Fructus". The new prescriptions extracted by entropy cluster analysis included 7 groups such as " Rehmannize Radix et Praeparata, Cistanches Herba, Corni Fructus and Asparagi Radix". Conclusion:The treatment of VD by TCM compounds with national patents is mainly based on tonifying deficiency, promoting blood circulation and removing blood stasis, eliminating phlegm and dampness, expelling wind and dredging collaterals, opening orifices and resuscitation, which can provide reference for clinical practice and new drug research and development.

Objective: To construct 3β-HSD gene shRNA lentivirus interference vecto, then transfect into human MCF-7 cells, and construct cell line with 3β-HSD gene silencing, finally to study the effects of 3β-HSD on apoptosis induced by di- (2-ethylhexyl) phthalate (DEHP) . Methods: According to the mRNA sequence of 3β-HSD gene provided by GenBank, three interference sequences were designed and connected to PLVX-shRNA2-puro after annealing. The recombinant lentivirus vector was transfected into 293FT cells, the virus supernatants were collected and infected with MCF-7 cells. After puromycin screening, MCF-7 cells with 3β-HSD gene silencing were constructed. The cells with 3β-HSD gene silencing were identified by real-time quantitative PCR and western blot. Then the 3β-HSD gene silencing cells and MCF-7 cells were treated at various doses of DEHP for 24 hours to detect the gene expression and protein expression of apoptosis genes including Bax, Caspase-3 and Caspase-8. Results: The interference sequence of 3β-HSD gene inserted into lentivirus vector PLVX-shRNA2-puro is consistent with the designed sequence. 3β-HSD gene expression level in MCF-7 cells with 3β-HSD gene silencing was 77% lower than than that of control MCF-7 cells. 3β-HSD protein level in MCF-7 cells with 3β-HSD gene silencing was 74% lower than that of control MCF-7 cells. After DEHP treatment in MCF-7 cells with 3β-HSD gene silencing and control MCF-7 cells, qRT-PCR results showed that Bax gene expression levels increased by 28%-54%, Caspase-3 gene increased by 13%-49%, Caspase-8 gene increased by 21%-70% in MCF-7 cells when compared with the control group. Additionally, in the 3β-HSD gene silencing cells, Bax gene expression level decreased by 11%-28%, Caspase-3 gene expression decreased by 12%-23%, Caspase-8 gene expression decreased by 11%-34%, compared with the same treatment group of MCF-7 cells. Western blot results showed that Bax protein expression level increased by 28%-61%, Caspase-3 protein expression level increased by 40%-48%, Caspase-8 protein increased by 31%-84% in MCF-7 cells when compared with the control group. In 3β-HSD gene silencing cells, Bax protein expression level increased by 11%-27%, Caspase-3 protein increased by 21%-40%, Caspase-8 protein increased by 12%-25%, compared with the same treatment group of MCF-7 cells. Conclusion The stable 3β-HSD gene silencing cell line are successfully constructed in this study. DEHP can induce increased expression of apoptotic gene and protein. Silencing of 3β-HSD gene can inhibit the activation of apoptotic gene by DEHP in a certain degree.

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.