OBJECTIVE: To optimize the extraction and purification technology of flavonoids from Morus alba.METHODS: The extraction technology was optimized by orthogonal test and flavonoids were filtered through AB-8 macroporous resin.RESULTS: The optimal extraction technology was as follows: 20 fold 60% ethanol,extracting for 40 min,at 80℃ with ultrasonic wave.CONCLUSION: Established extraction technology of total flavonoids has good extraction rate,which provided a theoretical basis for industvial production.

Objective To investigate the application value of extracorporeal extended field radiotherapy in locally advanced cervical cancer. Methods A total of one hundred and twenty patients with stage IIB?IVA cervical cancer in the First Affiliated Hospital of Hebei North University from June 2012 to June 2014 were randomly divided into two groups:the control group and the observation group,each with 60 cases. The routine concurrent chemoradiotherapy was adopted in the control group. The observation group was treated with extracorporeal extended field radiotherapy combined with concurrent chemotherapy. The patients have been followed up for three years. The total efficacy rate,toxicity reaction,local progression free survival time ( PFS) and survival rate of the two groups were compared. Results The total efficacy rate in the observation group was significantly higher than that of the control group ( 88. 3%( 53/60 ) vs. 73. 3%( 44/60 ) , χ2 = 4. 357, P=0. 037),while the incidences of toxicity reaction in the two groups were 18. 3%(11/60) and 16. 7%(10/60), the difference was not statistically significant (χ2=0. 058,P=0. 810) ,the percentages of I and II degree in the two groups were 3. 4%( 2/60 ) and 3. 4%( 2/60 ) , the difference between the two groups was not statistically significant (Z=0. 000,P=1. 000). The PFS value in the observation group was significantly longer than that of the control group ( 25. 6 months vs. 13. 8 months,χ2 = 25. 624, P= 0. 000 ) , and the survival rate in the observation group improved significantly ( 53. 3%( 32/60 ) vs. 33. 3%( 20/60 ) , the difference was statistically significant (χ2 = 4. 887, P= 0. 027 ) . Conclusion Extracorporeal extended field radiotherapy is safe and effective in the treatment of locally advanced cervical cancer.

Objective To investigate the relationship between prothrombin fragment 1+2 (F1+2) and peripherally inserted central catheter (PICC) associated thrombosis in cervical cancer patients, and provide certain clinical basis of early prevention in peripherally inserted central catheter associated thrombosis in cervical cancer patients. Methods One hundred and forty cervical patients with PICC were enrolled in this study, and they were divided into thrombosis group (35 patients) and non-thrombosis group (105 patients). The level of F1+2 was examined using enzyme-linked immunoassay, and was analyzed according to the clinic features. Results The level of F1+2 was correlated with clinical stage (r = 0.640, P = 0.004);but was not correlated with age, type of tumor and concurrent radiochemotherapy (P>0.05). The level of F1+2 in thrombosis group was (520.343 ± 121.759) pmol/L, in non- thrombosis group was (388.361 ± 104.873) pmol/L, and there was significant difference (P =0.001). The multi-factors Logistic analysis showed that the level of F1+2 (OR=1.011, P=0.001) and age (OR = 21.025, P = 0.031) were independent risk factors for the PICC associated with thrombosis in cervical cancer. Conclusions The level of F1+2 is closely related with clinical stage and PICC associated thrombosis, and it is an independent risk factor for the PICC associated with thrombosis in cervical cancer.

Objective:To study the characteristics of tumor microvascular perfusion by contrast-enhanced ultrasound (CEUS) in patients with breast cancer, and to analyze its relationship with pathology.Methods:The clinical data of 180 breast cancer patients admitted to Tangshan People′s Hospital from February 2019 to March 2021 were retrospectively analyzed. They were examined by contrast-enhanced ultrasound before surgery, and the specimens were sent for pathological biopsy after surgery. The characteristics of tumor microvascular perfusion under CEUS were observed, and the correlation between the characteristics and pathological classification and grade were analyzed.Results:The results of the CEUS showed that the contrast agentrapid infusion was 47.78%(86/180), slowly filled was 60.00%(108/180), the mass showed hyperenhancement when the contrast agent reached its peak was 42.78%(77/180), the contrast agent slowly withdrew was 42.78% (77/180), and the contrast agent retention in clearance was 65.56% (118/180). Pathological biopsy revealed that among 180 patients, 16 patients (8.89%) were non-invasive carcinoma, 41 patients (22.78%) were invasive lobular carcinoma, 88 patients (48.89%) were invasive ductal carcinoma, 10 patients (5.56%) were mucinous adenocarcinoma, 11 patients (6.11%) were medullary carcinoma, 8 patients (4.44%) were squamous carcinoma, 6 patients (3.33%) were hard carcinoma (3.33%). There was no correlation between tumor microvascular perfusion characteristics and pathological classification under CEUS ( P>0.05). Pathological biopsy showed that 95 patients (52.78%) were grade Ⅰ, 49 patients (27.22%) were gradeⅡand 36 patients (20.00%) were grad Ⅲ. There was a certain correlation between tumor microvascular perfusion characteristics and pathological grade under CEUS ( P<0.05). Conclusions:There is a certain relationship between tumor microvascular perfusion characteristics detected by CEUS and pathological grading in patients with breast cancer. Analysis of the microvascular perfusion characteristics can provide an important basis for pathological grading.

OBJECTIVETo investigate whether down-regulation of Twist1 could change sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol.

METHODSHNE1 cells were transfected with the small interfering RNA (siRNA) expression vector pSuppressor-Retro-Si-Twist, containing the short hairpin RNA (shRNA) sequence targeting the Twist gene-coding region by Fugene 6. Positive clones were then selected in Neomycin (400 microg/ml) for 21 days. The low expressions of Twist1 were examined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot. Drug sensitivity of si-Twist1 HNE1 to taxol was determined by Annexin V-fluorescein isothiocyanate( FITC)/propidium lodide (PI) double-labeled flow cytometry and detection of DNA ladder. The Effect of Twist1 inactivation on HNE1 cell proliferation was observed by MTT assay and flow cytometry.

RESULTSAnnexin V- FITC-PI assay showed that apoptosis ratio was 40.2% in si-Twist HNE1 after treated with 10 ng/ml taxol, significantly higher than that in the control siRNA group 24.3%. The deference had statistic meaning. After the re-expression of HNE1, apoptosis ratio was 44.80% +/- 4.80% (x +/- s) in low Twist1 protein expression group and that was 27.00% +/- 2.91% in high expression group. The deference had statistic meaning (t = 4.374, P = 0.049). Real time PCR test revealed apoptosis protein bcl-2 expression in si-Twist HNE1 was 0.28 +/- 0.05, significantly lower than that in the control siRNA HNE1 (0.57 +/- 0.08, t = 6.710, P = 0.021), nevertheless, significant bax and bcl-XL changes were not observed (t = 2.000, P = 0.184 and t = 1.502, P = 0.272). MTT and FCM showed that down-regulation of Twist1 did not alter cell proliferation rate (P>0.05).

CONCLUSIONSDown-regulation of Twist1 could increase drug sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol by inducing apoptosis. These results suggested that Twist1 may be a promising treatment target for nasopharyngeal carcinoma therapy.

Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; Nuclear Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA Interference ; RNA, Small Interfering ; Twist-Related Protein 1 ; genetics

Objective To explore the effect of high-frequency repetitive transcranial magnetic stimulation (rTMS) on central facial paralysis after ischemic stroke. Methods From June, 2020 to June, 2021, 54 patients with central facial palsy after ischemic stroke who were hospitalized in the Rehabilitation Department of Xuzhou Central Hospital were randomly divided into control group (n = 27) and experimental group (n = 27). Both groups were given conventional rehabilitation treatment, including medication and facial muscle rehabilitation training. The experimental group was treated with 5 Hz rTMS on the affected primary motor cortex, and the control group was treated with the same parameters of sham stimulation at the same site. Before treatment and four weeks after treatment, the House-Brackmann Grading System 2.0 (HBGS-2), the Sunnybrook Facial Grading System, the horizontal distance difference between the bilateral mouth corners to the lower center of the philtrum at rest, the horizontal distance difference between the bilateral mouth corners to the intersection of the mandibular central incisor when showing the teeth at the best effort and the angle of the tongue midline deviating from the facial midline when the tongue was stretched out were used to evaluate the facial nerve function of the patient. Results One case dropped down in each group. Before treatment, there was no significant difference in the scores of HBGS-2 and Sunnybrook Facial Grading System, the horizontal distance difference between the bilateral mouth corners to the lower center of the philtrum at rest, the horizontal distance difference between the bilateral mouth corners to the intersection of the mandibular central incisor when showing the teeth at the best effort, and the angle of the tongue midline deviating from the facial midline when the tongue was stretched out between two groups (P > 0.05). After treatment, all the indexes significantly improved in both groups (|t| > 8.987, P < 0.001), and were better in the experimental group than in the control group (t > 2.939, P < 0.01). Conclusion 5 Hz rTMS on the affected primary motor cortex is effective on the facial nerve function of patients with central facial palsy after ischemic stroke.

OBJECTIVE: To investigate the effect of miR-4443 expression on migration and invasion of breast cancer. METHODS: We examined the expression of miR-4443 in breast carcinoma in situ and paired adjacent tissues from 3 breast cancer patients with high-throughput sequencing and verified the results using TCGA database. We also detected miR-4443 expressions using real-time quantitative PCR (RT-qPCR) in low invasive and highly invasive breast cancer cells (MCF-7 and MDA-MB-231 cells, respectively). The changes in apoptosis, migration and invasion of MCF-7 and MDA-MB-231 cells after transfection with miR-4443 mimics, mimics-NC, miR-4443 inhibitor or inhibitor-NC were analyzed using flow cytometry, wound healing assay and Transwell invasion assay. The target gene of miR-4443 was predicted by bioinformatics software and validated by a dual luciferase reporter gene system. RT-qPCR and Western blotting were performed to detect the expression of recombinant human phosphatidyl ethanolamine binding protein 1 (PEBP1) in the transfected cells. RESULTS: The expression of miR-4443 was significantly higher in the breast cancer tissues than in the adjacent tissues ( CONCLUSIONS MiR-4443 promotes the migration and invasion of breast cancer cells by inhibiting the expression of PEBP1, suggesting the possibility of suppressing miR-4443 expression as a potential therapeutic strategy for breast cancer.
Breast Neoplasms/genetics* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; MCF-7 Cells ; MicroRNAs/genetics* ; Neoplasm Invasiveness/genetics* ; Phosphatidylethanolamine Binding Protein

As a leading cause of respiratory disease, influenza A virus (IAV) presents a pandemic threat in annual seasonal outbreaks. Given the limitation of existing anti-influenza therapies, there remains to be a requirement for new drugs. Compound Yi-Zhi-Hao pellet (CYZH) is a famous traditional Chinese medicine (TCM) used in the clinic, whose formula has been recorded into treat common cold. In this study, we found that CYZH exhibited a broad-spectrum anti-influenza activity and inhibited the expression of viral RNA and proteins. Mechanistically, CYZH had no inhibitory activities against viral protein hemagglutinin and IAV RNA-dependent RNA polymerase. Instead, it induced activation of erythroid 2-related factor 2 (Nrf2) and nuclear factor kappa B (NF-B), which subsequently upregulated heme oxygenase-1 (HO-1) expression. Also, CYZH protected cells from oxidative damage induced by reactive oxygen series. In conclusions, CYZH inhibits IAV replication, at least partly by activating expression of the Nrf2/HO-1 pathway.

[Abstract] Objective: To construct and verify the anti-tumor activity of chimeric antigen receptor (CAR) modified NK-92 cells (CAR-NK-92 cells) targeting prostate stem cell antigen (PSCA) in cervical cancer. Methods: Lentiviral vector expressing CAR targeting PSCA was constructed, and PSCA CAR-NK-92 cells were obtained by lentivirus transfection. The expression of PSCA in human cervical cancer cells was determined by Flow cytometry and Western blotting. The killing effect of PSCA CAR-NK-92 cells against cervical cancer cells was verified by co-incubation of effector and target cells in vitro, and the tumor inhibitory ability of PSCA CAR-NK-92 cells was verified with the nude mice xenograft model in vivo. Results: PSCA CAR-NK-92 cells were successfully constructed. PSCA was highly expressed in human cervical cancer Hela and MS751 cells (all P<0.01). In vitro co-incubation results showed that PSCA CAR-NK-92 cells could lyse PSCA+ cervical cancer transplanted tumor in a dose-dependent manner. In vivo anti-tumor data showed that PSCA CAR-NK-92 cells significantly inhibited the growth of cervical cancer cells compared with NK-92 cells transfected with vehicle vectors (P<0.01). In addition, PSCA CAR-NK-92 cells could effectively infiltrate tumor tissues and promote the secretion of anti-tumor cytokines TNF-α and IFN-γ (all P<0.01). Conclusion: The CAR-NK-92 targeting PSCA shows good anti-tumor effect on PSCA+ tumor cells both in vitro and in vivo, and has potential to be a therapeutic strategy for cervical cancer.

Post-traumatic stress disorder (PTSD) is a psychiatric disease that seriously affects brain function. Currently, selective serotonin reuptake inhibitors (SSRIs) are used to treat PTSD clinically but have decreased efficiency and increased side effects. In this study, nasal cannabidiol inclusion complex temperature-sensitive hydrogels (CBD TSGs) were prepared and evaluated to treat PTSD. Mice model of PTSD was established with conditional fear box. CBD TSGs could significantly improve the spontaneous behavior, exploratory spirit and alleviate tension in open field box, relieve anxiety and tension in elevated plus maze, and reduce the freezing time. Hematoxylin and eosin and c-FOS immunohistochemistry slides showed that the main injured brain areas in PTSD were the prefrontal cortex, amygdala, and hippocampus CA1. CBD TSGs could reduce the level of tumor necrosis factor-