0.05) . Significant decreases of contents of 14 kinds of macro-elements and trace elements in purified water were found compared with those in water samples after activated carbon filtration (t = 2.496 9-5.261 7, F2 and showed significant dose-response relationship respectively (r = 0.980, P

Establishing the genetic transformation system of medicinal plant is important to study their functional genes. Based on the established regeneration system of Sophra alopecuroides, 6 factors of genetic transformation were optimized, that was the concentration of Agrobacterium tumefaciens, the infection time, the co-cultivation time of agrobacterium tumefaciensand S.alopecuroides callus, the preculture time of S.alopecuroides callus, the adding method ofacetosyringone (AS) and the concentration of AS, respectively. The results showed that a maximum genetic transformation efficiency of 83.33% was achieved with 15d-precultured of S.alopecuroides callus, which was infected by A600=0.9 A. tumefaciens for 15 minutes and then co-cultivated for 48 hours with 200 μmol•L-1AS. The promoter sequence (1 260 bp) of upstream SaLDC was cloned from S.alopecuroides genomic DNA (gene bank accession number: KY038928). The deletion fragment of SaLDC promoter with different length (310,594,765,924,1 260 bp) were ligated with the GUS reporter gene to form five plant expression vectors named P310,P594,P765,P924,P1260, which were then transferred into S.alopecuroides callus. The GUS transient expression showed that all 5 different deletion fragment of SaLDC promoter can drive the GUS gene expression in S. alopecuroides callus. The SaLDC promoter we cloned has high promoter activity, and they may facilitate its function analysis in the future.

BACKGROUND: Non-small cell lung cancer (NSCLC) is one of the most lethal malignancies worldwide. A novel Chinese medicine formula-01 (NCHF-01) has shown significant clinical efficacy in the treatment of NSCLC, but the mechanism of this formula in the treatment of NSCLC is not fully understood. The aim of this study is to investigate the molecular mechanism of NCHF-01 in inhibiting NSCLC. METHODS: Lewis lung cells (LLC) tumor bearing mice were established to detect the tumor inhibitory effect of NCHF-01. The morphological changes of tissues and organs in LLC tumor-bearing mice were detected by hematoxylin-eosin (HE) staining. NSCLC cells were treated by NCHF-01. The effects of cell viability and proliferation were detected by MTT and crystal violet staining experiment. Flow cytometry was used to detect cell cycle, apoptosis and reactive oxygen species (ROS). Network pharmacology was used to predict the mechanism of its inhibitory effect of NSCLC. Western blot and immunohistochemistry (IHC) were used to detect the expression of related proteins. RESULTS: NCHF-01 can inhibit tumor growth in LLC tumor-bearing mice, and has no obvious side effects on other tissues and organs. NCHF-01 could inhibit cell viability and proliferation, induce G2/M phase arrest and apoptosis, and promote the increase of ROS level. Network pharmacological analysis showed that NCHF-01 exerts anti-NSCLC effects through various biological processes such as oxidative stress and central carbon metabolism. NCHF-01 can reduce the protein expression and enzyme activity of the key enzymes 6-phosphate glucose dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) in the pentose phosphate pathway (PPP). CONCLUSIONS NCHF-01 can inhibit NSCLC through oxidative stress dependent on the PPP.
Animals ; Mice ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/pathology* ; Reactive Oxygen Species/therapeutic use* ; Medicine, Chinese Traditional ; Pentose Phosphate Pathway ; Oxidative Stress ; Cell Line, Tumor ; Cell Proliferation ; Apoptosis

Objective: To investigate whether type 2 diabetes mellitus increases the risk of hepatitis B-related cirrhosis combined with type 2 diabetes mellitus for the occurrence of primary hepatocellular carcinoma, and to compare the effects of different nature of diabetes duration on the risk of different anti-diabetic drugs. Methods: A retrospective case-control study was conducted. (1) 325 cases with hepatitis B-related cirrhosis complicated with primary hepatocellular carcinoma were selected as the study group and 601 patients with hepatitis B cirrhosis as the control group. The relationship between diabetes mellitus and the risk of primary liver cancer was analyzed by multivariate logistic regression analysis. (2) Selected the study group and control group combined with type 2 diabetes mellitus, and used multivariate logistic regression analysis to study the relationship between diabetes-related factors and the risk of primary liver cancer. Results: The incidence of diabetes was 14.2% in the study group and 6.0% in the control group, and the difference was statistically significant between the two groups (P < 0.05). Multivariate logistic regression analysis showed that type 2 diabetes was one of the independent risk factors for primary hepatocellular carcinoma, which had increased the risk of primary hepatocellular carcinoma (adjusted odds ratio (AOR): 1.982, 95% CI: 1.224-3.210). Patients with diabetes > 10 years (adjusted ratio: AOR value 6.011, 95% CI: 1.659-21.777) were at significantly higher risk for primary hepatocellular carcinoma than that of patients with diabetes < 10 years. Metformin (adjusted odds ratio: AOR 0.188, 95% CI: 0.052-0.688) had reduced the risk, while insulin (adjusted odds ratio: AOR 6.682, 95% CI: 1.899-23.510) had increased the risk. Conclusion Type 2 diabetes mellitus is one of the independent risk factors for primary HCC, which can increase the risk of hepatocellular carcinoma in hepatitis B cirrhosis in relation to the duration of diabetes mellitus. The risk of hepatocellular carcinoma is higher in patients with duration of diabetes > 10 years and metformin reduces the risk.

OBJECTIVE: To screen for differentially expressed circular RNAs (circRNAs) in the serum of preterm infants with intraventricular hemorrhage (IVH) and explore the competitive endogenous RNA (ceRNA) mechanism of circRNAs in IVH in these infants. METHODS: Fifty preterm infants (gestational age of 28 to 34 weeks) admitted in our department between January, 2019 and January, 2020 were enrolled in this study, including 25 with a MRI diagnosis of IVH and 25 without IVH. Serum samples were collected from 3 randomly selected infants from each group for profiling differentially expressed circRNAs using circRNA array technique. Gene ontology (GO) and pathway analyses were performed to reveal the function of the identified circRNAs. The circRNA-miRNA-mRNA network was constructed to identify the co-expression network of hsa_circ_ 0087893. RESULTS: A total of 121 differentially expressed circRNAs were identified in the infants with IVH, including 62 up-regulated and 59 down-regulated circRNAs. GO and pathway analyses showed that these circRNAs were involved in multiple biological processes and pathways, including cell proliferation, activation and death, DNA damage and repair, retinol metabolism, sphingolipid metabolism, cell adhesion molecules. Among these circRNAs, hsa_circ_0087893 was found to have significant down-regulation in IVH group and co-express with 41 miRNAs and 15 mRNAs (such as miR-214-3p, miR-761, miR-183-5p, AKR1B1, KRT34, PPP2CB, and HPRT1). CONCLUSION The circRNA hsa_circ_0087893 may function as a ceRNA and play an important role in the occurrence and progression of IVH in preterm infants.
Infant, Newborn ; Infant ; Humans ; RNA, Circular ; Infant, Premature ; MicroRNAs ; RNA, Messenger ; Cerebral Hemorrhage/genetics* ; Aldehyde Reductase

To explore the effectiveness and safety of endovascular recanalization for chronic internal carotid artery occlusion patients under Hasan classification.To evaluate the guiding value of Hasan classification for endovascular recanalization for chronic internal carotid artery occlusion.To investigate the effect of successful endovascular recanalization on systolic blood pressure.Methods We retrospectively reviewed 30 consecutive chronic internal carotid artery occlusion patients.According to Hasan classification and DSA results of our hospital，the study subjects were divided into 4 types：A，B，C and D.After admission，the rate of recanalization，perioperative complication and followup outcomes were collected.Results Among the 30 CICAO patients，83.3% were male and 16.7% were female，with an average age of （58.5±8.6） years.The success rate of endovascular recanalization in 30 patients was 63.3%.According to Hasan classification，the success rate of endovascular recanalization in type A was 81.8%，Type B 75%，Type C 50%，Type D 0%；The perioperative complications was 23.3%，9.1% for type A and 37.5% for type B，Type C 37.5%，Type D 0%，no severe stroke or death occurred during perioperative period.No related complications occurred in all patients during followup.There was statistical significance in systolic blood pressure before and after interventional recanalization in successful group （P<0.01）.There was no significant difference in preoperative and postoperative systolic blood pressure between the unsuccessful group of interventional recanalization （P>0.05）.Conclusion Interventional recanalization for CICAO patients is safe and effective according to Hasan classification criteria.This classification can screen patients suitable for interventional recanalization，predict the success rate of surgery.Successful interventional recanalization may result in a decrease in postoperative systolic blood pressure.

BACKGROUND:There are many methods in the clinic to treat pelvic fractures, mainly conservative treatment, internal fixation and external fixation. Conservative treatment often causes complications due to poor reduction after fractures. Fixation has good effects on repair of unstable fractures, but fixation is seldom used for pelvic fractures.
OBJECTIVE:To observe the effects of internal fixation on unstable pelvic fractures, and compare with conservative treatment and external fixation.
METHODS:126 cases of unstable pelvic fractures from Longhua District People’s Hospital of Shenzhen City from January 2008 to June 2014 were divided into three groups:conservative treatment group, external fixation group and internal fixation group (n=42). After treatment, patients received X-ray examination. Lindahl imaging criteria were used as evidence. The quality of fracture reduction was evaluated. Patients were regularly fol owed up after treatment. The recovery of limb function was evaluated according to Majeed standard. Repair effects, the excel ent and good rates of fracture healing and cal us growth were evaluated in the last fol ow-up.
RESULTS AND CONCLUSION:During the last fol ow-up, the total efficiency was 81%in the internal fixation group, 69%in the conservative treatment group, and 71%in the external fixation group, and results were significantly better in the internal fixation group than in the other two groups (P<0.05). The Lindahl and Majeed scores were significantly higher in the internal fixation group than in the other two groups (P<0.05). These results suggest that internal fixation for unstable pelvic fracture obtained better recovery effects and efficiency than conservative treatment and external fixation. Thus, the internal fixation is more suitable for patients with unstable pelvic fractures.

This study aims to investigate the effects of Blaps rynchopetera Fairmaire and/or cyclophosphamide on the proliferation and apoptosis of lung cancer cells and decipher the underlying mechanism. B. rynchopetera and cyclophosphamide-containing serum and blank serum were prepared from SD rats. Cell counting kit-8(CCK-8) assay was employed to examine the proliferation of lung cancer cell lines A549 and Lewis treated with corresponding agents. The Jin's formula method was used to evaluate the combined effect of the two drugs. According to the evaluation results, appropriate drug concentrations and lung cancer cell line were selected for subsequent experiments, which included control, B. rynchopetera, cyclophosphamide, B. rynchopetera + cyclophosphamide, and B. rynchopetera + Wnt/β-catenin pathway agonist lithium chloride(LiCl) groups. Immunocytochemistry was employed to measure the expression of proliferation-related proteins in Lewis cells after drug interventions. Flow cytometry was employed to determine the cell cycle and apoptosis. The expression levels of proliferating cell nuclear antigen(PCNA), cyclinD1, B-cell lymphoma 2(Bcl-2), Bcl-2-assiocated X protein(Bax), Wnt1, and β-catenin were determined by Western blot. The results showed that B. rynchopetera and/or cyclophosphamide significantly inhibited the proliferation of A549 and Lewis cells. Compared with B. rynchopetera alone, the combination increased the inhibition rate on cell proliferation. The combination of B. rynchopetera and cyclophosphamide demonstrated a synergistic effect according to Jin's formula-based evaluation. Compared with the control group, the B. rynchopetera, cyclophosphamide, and B. rynchopetera + cyclophosphamide groups showed increased proportion of Lewis cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the cyclophosphamide group, the combination group showed increased proportion of cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the B. rynchopetera group, the B. rynchopetera + LiCl group had deceased proportion of cells in G_0/G_1 phase, decreased apoptosis rate, down-regulated expression of Bax, and up-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. The results indicated that B. rynchopetera could inhibit the proliferation, arrest the cell cycle, and induce the apoptosis of lung cancer cells by inhibiting the Wnt/β-catenin signaling pathway. Moreover, B. rynchopetera had a synergistic effect with cyclophosphamide.
Rats ; Animals ; Wnt Signaling Pathway ; Lung Neoplasms/genetics* ; beta Catenin/metabolism* ; Proliferating Cell Nuclear Antigen ; bcl-2-Associated X Protein/metabolism* ; Rats, Inbred Lew ; Rats, Sprague-Dawley ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Cell Proliferation ; Cyclophosphamide ; Cell Line, Tumor

OBJECTIVEThe study aimed to develop the assay of chrysosplenetin (CHR), a metabolic inhibitor of artemisinin by UPLC-MS/MS in rat plasma and investigate the pharmacokinetics parameters of CHR.

METHODThe plasma samples were precipitated by acetonitrile to remove the proteins. Separation was carried out on a Shim-pack XR-ODS C,18(2. 0 mm x 100 mm, 2. 2 micromp.m) column using a mobile phase containing methanol-0. 1% formic acid (87:13) using by diazepam as internal standard. Mass spectrometer with electrospray ionization (ESI) operated in the positive ion mode was used for analysis. Total analysis time was 2 min.

RESULTThe assay was linear in the range 5-5 000 microg L-1 (r =0. 999 3) with recoveries in the range from 69. 0% to 81.2% and satisfied inter-, intra- precision and accuracy. CHR after oral administration is not easy to absorb with double or multimodal peak phenomenon. The t1/2 of CHR after intravenous injection was very short and that of low, medium, and high dosage was (17. 01 +/- 8. 06) , (24. 62 +/- 4. 59), (28. 46+/- 4. 63) min, respectively.

CONCLUSIONThe developed method was special, rapid, and sensitive for determination of CHR pharmacokinetics. [Key words] UPLC-MS/MS; chrysosplenetin; pharmacokinetics; plasma; rat

Animals ; Artemisinins ; antagonists & inhibitors ; Chromatography, High Pressure Liquid ; methods ; Female ; Flavonoids ; blood ; pharmacology ; Male ; Rats ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry

ObjectiveTo evaluate the effect of the integrated echinococcosis control program in Ningxia Hui Autonomous Region from 2011 to 2018. MethodsA package of integrated interventions were employed for echinococcosis control in 22 counties (districts) of Ningxia Hui Autonomous Region from 2011 to 2018, including screening of human echinococcosis, treatment of echinococcosis patients, deworming of domestic dogs and monitoring of infections, surveillance of echinococcosis in bovines and sheep, health education. The detection of human echinococcosis, seroprevalence of anti-Echinococcus antibody in children at ages of 6 to 12 years, the Echinococcus coproantigen-positive rate in domestic dogs, prevalence of echinococcosis in bovines and sheep, and the awareness of echinococcosis control knowledge were investigated and compared during the period between 2011 and 2018. Results The detection of human echinococcosis appeared a decline tendency in Ningxia Hui Autonomous Region over years during the period from 2011 to 2018 (χ²_trend = 82.22, P < 0.05), and the prevalence of human echinococcosis decreased from 0.31% in 2011 to 0.15% in 2018. The seroprevalence of anti-Echinococcus antibody appeared a decline tendency in children at ages of 6 to 12 years over years (χ²_trend = 439.64, P < 0.01), and the prevalence of anti-Echinococcus antibody decreased from 6.12% in 2011 to 0.67% in 2018. The Echinococcus coproantigen-positive rate appeared a decline tendency in domestic dogs over years (χ²_trend = 260.33, P < 0.05), and the prevalence of anti-Echinococcus antibody decreased from 7.11% in 2011 to 0.75% in 2018. The prevalence of bovine and sheep echinococcosis reduced from 3.26% and 5.08% in 2011 to 1.35% and 0.76% in 2018, and Echinococcus predominantly parasitized in bovine (92.00%) and sheep (93.94%) livers. A total of 63 stool samples were collected from red fox, wolf and badger in Xiji and Haiyuan counties of Ningxia Hui Autonomous Region in 2013 and 2014, with no Echinococcus coproantigen-positives detected, and 107 domestic cats and 3 domestic dogs were dissected in these two counties, with no Echinococcus found. A total of 6 046 wild mice were dissected in Xiji County, Yuanzhou District and Haiyuan County of Ningxia Hui Autonomous Region from 2016 to 2019, and the prevalence of E. multilocularis was 0.31%. The awareness of echinococcosis control knowledge appeared an increasing tendency in Ningxia Hui Autonomous Region over years from 2011 to 2018 (χ²_trend = 3 367.97, P < 0.01), and the awareness increased from 21.83% in 2011 to 72.24% in 2018. Conclusions The integrated echinococcosis control program achieves a remarkable effect in Ningxia Hui Autonomous Region, and the transmission of echinococcosis has been preliminarily controlled. However, the echinococcosis transmission risk remains in few regions, and the integrated echinococcosis control program remains to be reinforced.