ObjectiveTo observe the mechanism through which Qige San inhibits tumor angiogenesis in esophageal cancer （EC） by regulating the microRNA-133a （miR-133a）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodsCell experiments were conducted by transfecting miR-133a mimics through liposomes and utilizing PI3K/Akt signaling pathway inhibitors. The effects of 60 mg·L^-1 and 100 mg·L^-1 Qige San on inflammatory factor secretion in the supernatant of EC9706 cells （esophageal cancer cells） were observed. EC9706 cell supernatants were then extracted and used as conditioned media to treat human umbilical vein endothelial cells （HUVECs）. The effects of the conditioned media on the proliferation， migration， tube formation， and apoptosis of HUVECs were examined. Animal experiments involved establishing a subcutaneous tumor model in nude mice with EC9706 cells. After grouping and administration， tumor volumes were measured regularly. At the end of the experiment， tumors were harvested， and histopathological changes were observed under a microscope after hematoxylin-eosin （HE） staining. Real-time polymerase chain reaction （Real-time PCR） was adopted to measure miR-133a expression in the tumor tissue of each group， while Western blotting was employed to analyze the expression of key signaling molecules， such as phosphorylated Akt （p-Akt）， as well as endothelial cell markers， cluster of differentiation 31 （CD31） and cluster of differentiation 34 （CD34）， in the tumor tissue. ResultsCell experiments showed that compared with the blank group， the supernatant of EC9706 cells promoted tube formation of HUVECs （P<0.05）. Both low and high doses of Qige San inhibited tube formation of HUVECs （P<0.05）. All doses of Qige San， PI3K/Akt inhibitor， and the combination reduced the content of interleukin-6 （IL-6） （P<0.05）， while high-dose Qige San， inhibitor， and combination all reduced the content of interleukin-8 （IL-8） （P<0.05）. Compared with the EC9706 blank supernatant group， Qige San-conditioned medium inhibited the proliferation of HUVECs （P<0.05）. Qige San， the inhibitor， and the combination all inhibited the migration and tube formation （P<0.05） and increased the late apoptosis and total apoptosis rates of HUVECs （P<0.05）. In addition， the low-dose combination induced early apoptosis of EC9706 cells （P<0.05）. Compared with the blank supernatant group， the miR-133a mimic combined with Qige San synergistically reduced IL-6/IL-8 （P<0.05）， and enhanced the inhibition on proliferation， migration， and tube formation of HUVECs （P<0.05）. Animal experimental results showed that compared with the model group， both Qige San and cisplatin inhibited the growth of xenograft tumors in nude mice （P<0.05）， promoted tumor cell necrosis， and reduced capillary formation. High-dose Qige San upregulated miR-133a expression （P<0.05）. Moreover， compared with the model group， cisplatin， Qige San， and the combination all downregulated the expression of p-Akt， CD31， and CD34 （P<0.05）. ConclusionQige San upregulates miR-133a expression， inhibit PI3K/Akt signaling pathway activity， and improve the tumor inflammatory microenvironment to suppress the growth and tumor angiogenesis of HUVECs， thereby inhibiting esophageal cancer.

Objective:To investigate the efficacy of Qingruxiao granules combined with tamoxifen in the treatment of breast hyperplasia and its effect on serum hypoxia-inducible factor-alpha (HIF-α), angiopoietin-2 (Ang-2) and prolactin (PRL) levels. Methods:Ninety-eight patients with breast hyperplasia admitted to Xi'an No.3 Hospital from June 2020 to January 2022 were retrospectively included in this study. They were divided into control and observation groups ( n = 49/group) according to different treatments. The control group was treated with tamoxifen alone. The observation group was treated with Qingruxiao granules combined with tamoxifen. Clinical efficacy, symptom score, ultrasound parameters (glandular layer thickness, longest diameter of mass, maximum diameter of hypoechoic area, inner diameter of lactating tube), endocrine hormone levels (estradiol, progesterone, and prolactin), HIF-α, and Ang-2 pre- and post-treatment, as well as the incidence of adverse reactions were compared between the two groups. Results:Total response rate in the observation group was significantly higher than that in the control group [93.88% (4/49) vs. 77.55%, χ2 = 5.33, P < 0.05). After treatment, breast mass score, breast pain, systemic accompanying symptom, and nipple discharge in the observation group were (1.34 ± 0.29) points, (1.02 ± 0.36) points, (0.68 ± 0.17) points, (0.97 ± 0.15) points, respectively, which were significantly lower than (1.57 ± 0.23) points, (1.45 ± 0.41) points, (0.95 ± 0.26) points, and (1.28 ± 0.26) points, respectively, in the control group ( t = 4.35, 5.52, 6.08, 7.23, all P < 0.001). The glandular layer thickness, the longest diameter of mass, the maximum diameter of hypoechoic area, and the inner diameter of lactating duct in the observation group were (9.45 ± 1.67) mm, (11.46 ± 3.68) mm, (14.37 ± 4.22) mm, and (1.23 ± 0.39) mm, respectively, which were significantly lower than (11.26 ± 2.51) mm, (16.33 ± 4.01) mm, (19.87 ± 5.01) mm, (1.54 ± 0.48) mm in the control group ( t = 4.20, 2.26, 5.88, 3.51, all P < 0.001). Serum estradiol and prolactin levels in the observation group were (122.35 ± 29.76) ng/L and (205.64 ± 36.42) IU/L, respectively, which were significantly lower than (139.76 ± 30.48) ng/L and (251.49 ± 41.87) IU/L in the control group ( t = 2.86, 5.78, both P < 0.05). Serum progesterone level in the observation group was (9.22 ± 1.57) μg/L, which was significantly higher than (7.18 ± 1.21) μg/L in the control group ( t = -7.20, P < 0.05). Serum HIF-α and Ang-2 levels in the observation group were (0.15 ± 0.05) ng/L and (0.98 ± 0.11) ng/L, respectively, which were significantly lower than (0.24 ± 0.07) ng/L and (1.49 ± 0.22) ng/L in the control group ( t = 7.32, 14.51, both P < 0.001). There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusion:Qingruxiao granules combined with tamoxifen can effectively improve clinical symptoms, reduce tumor size, regulate endocrine hormone levels, decrease the expression of angiogenic factors in patients with breast hyperplasia, and is highly safe.

Objective:To explore the mechanism of the extraction of Carthami flos in the treatment of chronic alcoholic liver injury based on metabolomics. Methods:The rats were randomly divided into normal group, model group, positive control group and low, medium and high dose groups of the extraction of Carthami flos, with 10 rats in each group. In addition to the normal group, the other groups were gavaged with 8 ml/kg of 56° Niulanshan Baijiu to prepare the rat model with chronic alcoholic liver injury. After the successful modeling, the positive control group was gavaged with 0.36 mg/kg of Hugan tablet, and the low, medium and high dose groups were gavaged with 0.476 7, 1.430 1 and 4.290 3 g/kg of the extraction of Carthami flos respectively, once a day for 21 days. The contents of GPT, GOT, TG and ALP2 in serum were detected by automatic biochemical analyzer, and the effects of the extraction of Carthami flos on chronic alcoholic liver injury in rats were analyzed by Ultra High Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS), cluster analysis, Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA). Results:The levels of GPT [(42.11±6.58)U/L, (42.38±6.58)U/L vs. (49.96±10.70)U/L] and GOT [(104.81±14.70)U/L, (102.91±23.65)U/L vs. (159.66±53.69)U/L] decreased ( P<0.05 or P<0.01), while the levels of TG [(0.85±0.29)U/L, (0.85±0.23)U/L vs.(0.62±0.21)U/L] and ALP2 [(104.53±13.53)U/L, (100.30±17.28)U/L vs.(77.13±12.54)U/L] increased ( P<0.05 or P<0.01); The results of cluster analysis, PCA and OPLS-DA showed that the model group and the high-dose group of the extraction of Carthami flos could be distinguished obviously. A total of 20 chemical markers were obtained in the serum of rats with chronic alcoholic liver injury treated with the extraction of Carthami flos. Among them, the extraction of Carthami flos can down regulate the level of serum linolenic acid triglyceride in rats with chronic alcoholic liver injury and up regulate triglyceride, palmitic acid, lysophosphatidylcholine, palmitoyl ethanolamide, epinephrine, sphingosine, lysophosphingomyelin α-Linolenic acid, docosahexaenoic acid and eicosapentaenoic acid are 10 endogenous metabolites. Conclusion:The treatment of chronic alcoholic liver injury with the extraction of Carthami flos may be related to the regulation of endogenous metabolites docosahexadilute acid, docosapentadilute acid α- Linolenic acid.

Objective:To investigate the relationship between NOD-like receptors 3 (NLRP3) inflammasomes mediated inflammatory response and the pathogenesis of depression, and to investigate the effect of fluoxetine on this process.Methods:120 male C57BL/6J (wild-type) mice were randomly divided into control group 1, control group 2, chronic unpredictable mild stress (CUMS) group, and CUMS plus fluoxetine group. Another 30 male C57BL/6J (NLRP3-/-) mice were selected as NLRP3-/- group. Control group 1 and control group 2 had No treatment; CUMS group, CUMS plus fluoxetine group, and NLRP3-/- group were given chronic unpredictable mild stimulation for six weeks. After modeling, mice in control group 2 and CUMS plus fluoxetine group were intraperitoneally injected with fluoxetine (10 mg/(kg·d)). In contrast, mice in the other three groups were injected with the same amount of normal saline every day for four weeks. Behavioral tests were performed once a week before and after stress stimulation. Tail venous blood was drawn immediately before stress stimulation, three weeks later and six weeks after stress stimulation and was centrifuged at 3000 r/min for 10 min. The supernatant was kept and frozen for future use. Serum levels of interleukin-1β(IL-1β) and interleukin-18 (IL-18) were detected by enzyme-linked immunosorbent assay. After drug intervention (10 mg/(kg·d) fluoxetine or the same volume of normal saline), the mice in each group underwent behavioral tests once a week. The results included the sugar water preference test, forced swimming test (FST), and tail suspension test (TST). Tail venous blood was drawn from mice in each group at 1, 3, and 4 weeks after fluoxetine administration, and the supernatant was centrifuged and stored for later use. Serum levels of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay. At the above time points, 5 mice in each group were sacrificed each time, and fresh hippocampal tissues were collected and stored at a low temperature. NLRP3, urinary winter peptidase (caspase-1), the induction of transcription factors-KB (nuclear factor-κB, NF-κB), IL-1β and the IL-18 in the hippocampus brain regions were detected by using Western Blot.Results:(1) Model establishment: After six weeks of CUMS, compared with control group 1 and control group 2, the sugar water consumption of mice in CUMS group and CUMS+fluoxetine group was significantly reduced, and the immobility time of FST and TST was significantly prolonged, which proved that the model establishment was successful. After CUMS, NLRP3-/- group mice did not show depression-like changes in FST, sugar water preference test, and TST, which indicated that the model failed to be established. (2) After intraperitoneal fluoxetine injection, there were no significant differences in sugar water consumption, FST and TST immobility time between control group 2 and control group 1 ( P>0.05), and the sugar water consumption of mice in CUMS plus fluoxetine group was significantly increased, compared with CUMS group ( P<0.05). The immobility time of FST and TST was significantly shortened ( P<0.05). (3) The results of the enzyme-linked immunosorbent assay showed that after stimulation of CUMS, compared with control group 1 and control group 2, the serum levels of IL-1β and IL-18 in CUMS group and CUMS plus fluoxetine group were significantly increased ( P<0.05), while NLRP3-/- group had no significant change ( P>0.05). After fluoxetine administration, the serum levels of IL-1β and IL-18 in CUMS plus fluoxetine group were significantly lower than those in CUMS group ( P<0.05). (4) Western blotting results showed that after stimulation of CUMS, compared with control group 1 and control group 2, mice brain hippocampus NLRP3 expression, caspase-1 activation and induction of transcription factors-κB (nuclear factor-κB, NF-κB), IL-1β, IL-18 expression significantly increased ( P<0.05) in CUMS and CUMS plus fluoxetine group. After fluoxetine treatment, mice brain hippocampus NLRP3 expression, caspase-1 activation, NF-κB, IL-1β, and IL-18 expression in CUMS plus fluoxetine group had a significant reduction (restored to control 1 group by 99%, 91%, 97%, 95%, and 97% respectively). Conclusion:(1) CUMS may bring more NLRP3, caspase-1, NF-κB, IL-1β, and IL-18 expression in mice hippocampus, which cannot be seen, in the NLRP3 gene knockout mice; (2) The fluoxetine treatment may significantly decrease the NLRP3, caspase-1, NF-κB, IL-1β, IL-18 expression on depressive model mice, and improve depressive behavior.

Objective:To investigate the relationship between NOD-like receptors 3 (NLRP3) inflammasomes mediated inflammatory response and the pathogenesis of depression, and to investigate the effect of fluoxetine on this process.Methods:120 male C57BL/6J (wild-type) mice were randomly divided into control group 1, control group 2, chronic unpredictable mild stress (CUMS) group, and CUMS plus fluoxetine group. Another 30 male C57BL/6J (NLRP3-/-) mice were selected as NLRP3-/- group. Control group 1 and control group 2 had No treatment; CUMS group, CUMS plus fluoxetine group, and NLRP3-/- group were given chronic unpredictable mild stimulation for six weeks. After modeling, mice in control group 2 and CUMS plus fluoxetine group were intraperitoneally injected with fluoxetine (10 mg/(kg·d)). In contrast, mice in the other three groups were injected with the same amount of normal saline every day for four weeks. Behavioral tests were performed once a week before and after stress stimulation. Tail venous blood was drawn immediately before stress stimulation, three weeks later and six weeks after stress stimulation and was centrifuged at 3000 r/min for 10 min. The supernatant was kept and frozen for future use. Serum levels of interleukin-1β(IL-1β) and interleukin-18 (IL-18) were detected by enzyme-linked immunosorbent assay. After drug intervention (10 mg/(kg·d) fluoxetine or the same volume of normal saline), the mice in each group underwent behavioral tests once a week. The results included the sugar water preference test, forced swimming test (FST), and tail suspension test (TST). Tail venous blood was drawn from mice in each group at 1, 3, and 4 weeks after fluoxetine administration, and the supernatant was centrifuged and stored for later use. Serum levels of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay. At the above time points, 5 mice in each group were sacrificed each time, and fresh hippocampal tissues were collected and stored at a low temperature. NLRP3, urinary winter peptidase (caspase-1), the induction of transcription factors-KB (nuclear factor-κB, NF-κB), IL-1β and the IL-18 in the hippocampus brain regions were detected by using Western Blot.Results:(1) Model establishment: After six weeks of CUMS, compared with control group 1 and control group 2, the sugar water consumption of mice in CUMS group and CUMS+fluoxetine group was significantly reduced, and the immobility time of FST and TST was significantly prolonged, which proved that the model establishment was successful. After CUMS, NLRP3-/- group mice did not show depression-like changes in FST, sugar water preference test, and TST, which indicated that the model failed to be established. (2) After intraperitoneal fluoxetine injection, there were no significant differences in sugar water consumption, FST and TST immobility time between control group 2 and control group 1 ( P>0.05), and the sugar water consumption of mice in CUMS plus fluoxetine group was significantly increased, compared with CUMS group ( P<0.05). The immobility time of FST and TST was significantly shortened ( P<0.05). (3) The results of the enzyme-linked immunosorbent assay showed that after stimulation of CUMS, compared with control group 1 and control group 2, the serum levels of IL-1β and IL-18 in CUMS group and CUMS plus fluoxetine group were significantly increased ( P<0.05), while NLRP3-/- group had no significant change ( P>0.05). After fluoxetine administration, the serum levels of IL-1β and IL-18 in CUMS plus fluoxetine group were significantly lower than those in CUMS group ( P<0.05). (4) Western blotting results showed that after stimulation of CUMS, compared with control group 1 and control group 2, mice brain hippocampus NLRP3 expression, caspase-1 activation and induction of transcription factors-κB (nuclear factor-κB, NF-κB), IL-1β, IL-18 expression significantly increased ( P<0.05) in CUMS and CUMS plus fluoxetine group. After fluoxetine treatment, mice brain hippocampus NLRP3 expression, caspase-1 activation, NF-κB, IL-1β, and IL-18 expression in CUMS plus fluoxetine group had a significant reduction (restored to control 1 group by 99%, 91%, 97%, 95%, and 97% respectively). Conclusion:(1) CUMS may bring more NLRP3, caspase-1, NF-κB, IL-1β, and IL-18 expression in mice hippocampus, which cannot be seen, in the NLRP3 gene knockout mice; (2) The fluoxetine treatment may significantly decrease the NLRP3, caspase-1, NF-κB, IL-1β, IL-18 expression on depressive model mice, and improve depressive behavior.

Protein and Enzyme Engineering is the core and required course for colleague students majored in biotechnology, which plays an important role in the professional training system. In accordance with the "Guidelines for the Development of Ideological Education in Higher Education Institutions" issued by the Ministry of Education, we explored the combination of course teaching with ideological education by considering the features of the biotechnology major and the course and setting up rational teaching objectives. This paper described the strategy, design, implementation and evaluation approaches that were used in the course of "Protein and Enzyme Engineering" to achieve a good integration. The practice starts from story-telling, discussion of life, case study, hot issues discussion, literature discussion and presentations. The scientific spirits, civic character, global vision, eco-civilization and legal consciousness, as well as their native land emotion and cultural confidence, were boosted. The natural integration of the ideological education into the whole process of this course helped to better achieve the goal of curriculum education while promoting teaching excellence.
Biotechnology ; Curriculum ; Humans ; Students ; Universities

Objective:To explore the attitudes of parents of critically ill children to resuscitation companionship and its influencing factors so as to provide a scientific basis for improving the experience of children and their families in the rehabilitation process.Methods:A cross-sectional survey method was used, and parents of critically ill children who were hospitalized in Children's Hospital Affiliated to Zhengzhou University from August 2018 to August 2019 were selected by convenience sampling. The general data questionnaire and Family Presence During Resuscitation Benefits-Risks (FPDR-BRS) were used to investigate the attitudes of parents of critically ill children to the resuscitation companion and influencing factors were analyzed. A total of 200 questionnaires were distributed, 189 of which were effective, with an effective recovery rate of 94.5%.Results:The total scores of FPDR-BRS of 189 parents of critically ill children were (74.75±22.54) points. Univariate analysis showed that the total scores of FPDR-BRS of parents of critically ill children had statistically significant differences in the relationship with children, parents' occupations, cultural level and presence or absence of resuscitation experience ( P<0.05) . Multiple linear regression analysis showed that the relationship with children, parents' occupation and cultural level were the influencing factors of attitude to resuscitation companionship ( P<0.05) . Conclusions:Parents of critically ill children have a moderate level of recognition for resuscitation companionship. The relationship with children, parents' occupation and cultural level are factors that influence the attitude to resuscitation companionship. Medical staff should formulate targeted interventions based on the influencing factors of attitude to resuscitation companionship so as to improve the experience of children and their families during the resuscitation process.

Objective To study the effect of menopause status and the time of taking tamoxifen (TAM) on endometrial lesions after breast cancer surgery. Methods A total of 330 patients with postoperative vaginal irregular bleeding after breast cancer surgery or endometrial lesions after B ultrasonic from August 2007 to August 2017 in Northern Jiangsu People 's Hospital were retrospectively analyzed, including 180 cases of taking TAM treatment (medicine-taking group), and 150 cases of not taking TAM treatment (non medicine-taking group). The patients were also divided into the menopause group and the premenopausal group. According to the time of taking TAM, the patients were divided into < 2 years group, 2-5 years group and > 5 years group. Chi-square and Fisher test were used to compare the differences. Results The endometrial lesions incidence in the medicine-taking group was higher than that in the non medicine-taking group [84.44 ％ (152/180) vs. 56.00％(84/150);χ2=51.701, P=0.000]. The endometrial lesions rate in the menopause group was higher than that in the premenopause group [medicine-taking group: 69.70 ％ (46/66) vs. 92.98 ％ (106/114), χ2= 17.254, P= 0.000; non medicine-taking group: 46.15 ％ (35/65) vs. 63.53 ％(54/85), χ2 = 4.513, P= 0.034]. For the patients in the menopause group and the premenopause group, the incidence of endometrial lesions for those who took medicine for>5 years [96.00％(48/50), 85.19％(23/27)] was higher than that in the<2 years group and 2-5 years group [78.26％(18/23), 42.86％(6/14);95.12％(39/41), 72.00％(18/25) respectively], and there were statistical differences (χ2=7.619, P=0.022;χ2= 8.070, P= 0.018). The menopause was not correlated with staging, muscular lawyer infiltration and lymph metastasis postoperative (P> 0.05), but with the type of endometrial cancer (P= 0.013); the length of taking medicine was related with the type of endometrial cancer and the lymph metastasis (P=0.027). With the prolonged time of medicine-taking for postmenopause patients, the incidence of type Ⅱendometrial cancer and positive rate of lymph metastasis were also increased. Conclusions Taking TAM after surgery for breast cancer patients increases the risk of endometrial lesions. The longer the patients take the medicine, the greater risk of the lesions take, and the worse the pathological, histological type and prognosis of endometrial carcinoma are, which is more obvious for postmenopausal women who take TAM for more than 5 years.

Objective To propose a new observation procedure of blood pressure after the surgery of fiducial implantation for cyber knife, using the statistical analysis of the measurements of blood pressure at different time after the surgery. Methods By taking 200 patients after the surgery of fiducial implantation, the usual blood pressures and the blood pressures at different time after the surgery were compared statistically. Results Based on the statistical analysis, the usual blood pressures were statistically different from the blood pressure immediately after surgery, as well as the blood pressure at 0.5 hours after surgery [ systolic pressure (131.60 ±16.86), (129.08 ±14.77) mmHg (1 mmHg=0.133 kPa) vs. (123.00 ±11.80) mmHg, t=2.897 and 2.308, P<0.01 or 0.05;diastolic pressure (83.49±10.01), (81.75±8.99) mmHg vs. (78.22±7.59) mmHg, t=2.901 and 2.066, P<0.01 or 0.05]. The blood pressures measured at the other time after surgeries were not statistically different from the usual blood pressures, P>0.05. Conclusions Measuring the blood pressures at the time of 0, 0.5, 3.5 hours after the surgery can not only assure the complete recovery of the patients after the surgery, but also greatly reduce the work of nurses.

DNA, Neoplasm ; Humans ; Image Cytometry ; Pleural Effusion, Malignant ; diagnosis