ObjectiveTo explore the effect and mechanism of Jianpi Xiaoai prescription （JPXA） in ameliorating the 5-fluorouracil （5-FU） resistance of colon cancer. MethodsA HCT116/5-FU resistant cell line was established. Different concentrations （10%， 15%， 20%） of JPXA-containing serum and drug-free serum were used for intervention， and 10% fetal bovine serum （10% FBS）， fibroblast growth factor receptor （FGFR） inhibitor （AZD4547）， and recombinant fibroblast growth factor 2 （FGF2） were set as the control groups. Sensitive HCT116 cells were used in the FGF2 group， while HCT116/5-FU cells were used in other groups. Drug resistance， the level of FGF2 in the cell culture medium， the mRNA level of FGF2 in cells， and the protein levels of FGF2/FGFR and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） were determined. The drug-resistant cells were transplanted into the axilla of nude mice to establish a tumor model. The modeled mice were allocated into model， JPXA （15 g·kg^-1）， 5-FU （0.02 g·kg^-1）， JPXA+5-FU （15 g·kg^-1+0.02 g·kg^-1）， AZD4547 （0.012 5 g·kg^-1）， and AZD4547+5-FU （0.012 5 g·kg^-1+0.02 g·kg^-1） groups. The tumor growth and the protein levels of FGF/FGFR and PI3K/Akt in each group were observed. ResultsThe survival rate of HCT116/5-FU cells decreased in all the JPXA groups with different concentrations. The cell survival rate was decreased most obviously in the 20% JPXA group. The level of FGF2 in the cell culture medium and the mRNA level of FGF2 in cells of each JXPA group decreased， and the decrease was the most significant in the 20% group （P<0.01）. HCT116/5-FU cells showed up-regulated protein levels of FGF2 and phosphorylated fibroblast growth factor receptor 1 （p-FGFR1）， but down-regulated protein level of FGFR1 （P<0.01）. JPXA down-regulated the expression of FGF2 and p-FGFR1 and up-regulated the expression of FGFR1 （P<0.05）. In addition， JPXA down-regulated the expression levels of phosphorylated protein kinase B （p-Akt） and phosphorylated mammalian target of rapamycin （p-mTOR）， while up-regulating the expression levels of Akt and Bcl-2-asociated death promoter （Bad） （P<0.05）. Animal experiments showed that the JPXA combined with 5-FU significantly inhibited the growth of drug-resistant tumors， reduced the protein levels of FGF2， p-FGFR1， phosphorylated phosphatidylinositol-3-kinase （p-PI3K）， p-Akt， and p-mTOR， and increased the expression of Bad. It indicated that JPXA can inhibit the FGF2/FGFR1 signaling in colon cancer and regulate PI3K/Akt and downstream signaling pathways. ConclusionJPXA can ameliorate the chemotherapy resistance of colon cancer through down-regulating FGF2 expression and inhibiting the activation of the PI3K/Akt signaling pathway.

Objective To establish a DNA quantitation method based on dithiothreitol(DTT)-crystal vio-let.Methods DTT was used to decolorize crystal violet,mixed with different concentrations of λ-DNA and salmon sperm DNA standard samples or concentration standard samples,and the absorbance was read at 595 nm wavelength by microplate reader,and compared with the results of ultraviolet absorbance method.DTT-crystal violet method and ultraviolet absorbance method were used to compare the concentration of plasmid samples and the concentration of genomic DNA samples of cervical exfoliated cells.The protein tolerance of the two methods was assessed by simulating protein contaminants with bovine serum albumin(BSA).Results In the quantification of λ-DNA and salmon sperm DNA,the DTT-crystal violet method had a robust linear correlation between the absorbance at 595 nm and DNA concentration(r2＞0.95),and the measured concentrations of the standard samples were not significantly different from the theoretical concentrations of the prepared standard samples(P＞0.05).There was no significant difference in the concentration of plasmid samples measured by DTT-crystal violet method and ultraviolet absorption method(P＞0.05).The concen-tration of DNA samples from cervical exfoliated cells measured by ultraviolet absorption method was positive-ly correlated with that by DTT-crystal violet method(r=0.94,P＜0.01).The concentration of the standard sample containing BSA 1 μg/μL measured by ultraviolet absorption method was higher than that of the con-trol sample,and the difference was statistically significant(P＜0.01),whereas the DTT-crystal violet method was not significantly affected(P＞0.05).Conclusion DTT-crystal violet method has obvious advantages over the existing DNA quantitation method,and is suitable for DNA quantitative analysis in scientific research and clinic.

Objective:To establish a method for the identification of Qianghuo (Notopterygii Rhizoma et Radix) dispensing granules with PCR for the identification of dispensing granules.Methods:The collected samples were identified by DNA bar code, and the methods of extracting genomic DNA from Notopterygii Rhizoma et Radix, Qianghuo standard decoction and dispensing granules were established. Specific differential primers were designed based on ITS2 sequence, and the PCR amplification system and reaction conditions were optimized.Results:The PCR amplification system and reaction conditions were determined. The target bands of about 216 bp were obtained by amplification of Notopterygii Rhizoma et Radix, Qianghuo standard decoction and dispensing granules, and there was no interference from counterfeit products and blank.Conclusions:The specific PCR identification method of Qianghuo (Notopterygii Rhizoma et Radix) dispensing granules is established, which has good specificity. The detection limit of Notopterygii Rhizoma et Radix is 0.91 ng, and the detection limit of dispensing granules is 7.36 ng, which also provides a reference for the identification of other kinds of TCM dispensing granules.

Objective:To establish a polymerase chain reaction(PCR)method to accurately discriminate the crude materials and aqueous extract of Zaocys dhumnades,Elaphe carinata,Elaphe meollendorff and Ptyas korros.Methods:Specific primers were designed using mitochondrial Cytb gene(CO1)as a target gene,and annealing temperature,number of cycles and the type of DNA polymerases were optimized.The mixed samples were detected by this method.Results:By this multiplex allele-specific PCR identification method,135,182,246 and 197 bp of specific fragments were amplified from DNA templates of Zaocys dhumnades,Elaphe carinata,Elaphe meollendorffi and Ptyas korros,respectively,following the conditions:cycle number of 35,annealing temperature of 62 ℃.The adulterants and the blank control showed no bands.The method could simultaneously and accurately identify the snake-derived components in the mixed samples.Conclusion:The method can be used to identify the samples of Zaocys dhumnades,Elaphe carinata,Elaphe meollendorffi and Ptyas korros simultaneously,accurately and rapidly,and is suitable for the identification of standard decoctiond and formula granules samples.

Objective:To explore the effects of different time of skin-to-skin contact on physical/mental status and breastfeeding in cesarean section parturient, so as to provide reference for promoting breastfeeding.Methods:A total of 87 cesarean section parturient and children were collected from Taihe Hospital of Traditional Chinese Medicine from February 2021 to February 2022 as research objects by convenient sampling method. They were divided into A group, B group and C group by random digits table method, each group were 29 cases. Skin-to-skin contact was carried out in the three groups while the parturient was awake, A group contact time were 30 min, B group were 60 min, C group were 90 min. The intervention effect was assessed by Short Form McGill Pain Questionnaire, Visual Analogue Scale for Anxiety (VAS-A), Infant Brestfeeding Assessment Tool (IBFAT) as well as breast feeding related indicators.Results:Finally, 81 parturient were included, with 27, 28, and 26 cases in A group, B group, and C group. The scores of visual analogous scale, pain grading index, presents pain intensity, VAS-A were (6.74 ± 1.48), (19.33 ± 2.02), (4.48 ± 1.16), (5.22 ± 1.05) points in A group and (6.93 ± 1.41), (20.11 ± 2.50), (4.93 ± 1.56), (5.46 ± 1.04) points in B group, lower than in C group (7.69 ± 1.19), (22.08 ± 2.31), (5.73 ± 1.08), (6.31 ± 1.57) points, the differences were statistically significant ( t values were 2.05-4.36, all P<0.05). The successful rate of first breastfeeding and the rate of exclusive breastfeeding at 42 d after delivery were 55.6%(15/27), 59.3%(16/27) in A group, lower than in B group 82.1%(23/28), 85.7%(24/28) and in C group 88.5%(23/26), 88.5%(23/26), the differences were statistically significant ( χ2 values were 4.55-7.07, all P<0.05). The lactation start time, duration of first breastfeeding and IBFAT total scores were (17.04 ± 3.98) h, (10.44 ± 3.03) min, (4.74 ± 1.64) points in A group, lower than in B group and C group (19.22 ± 3.12) h, (12.36 ± 3.18) min, (7.07 ± 1.78) points and (16.69 ± 3.97) h, (13.62 ± 2.38) min, (7.58 ± 1.40) points, the differences were statistically significant ( t values were 2.45-5.25, all P<0.05). Conclusions:Skin-to-skin contact for 60 min can promote breastfeeding and alleviate anxiety and pain degree of cesarean section parturient.

ObjectiveTo explore the effects of Baihu Jia Renshen Tang （BHRS） on the related molecules on the phosphatidylinositol-3-kinase/protein kinase B（PI3K/Akt）signaling pathway in the liver of MKR diabetic model mice. MethodThirty 6-week-old MKR mice were selected and fed on a high-fat diet for four weeks，followed by intraperitoneal injection of streptozotocin（STZ）for the diabetes model establishment. The model was properly induced in the case of the fasting blood glucose （FBG） of ≥11.1 mmol·L^-1. After modeling，the mice were randomly divided into a model group，a BHRS group （12.09 g·kg^-1·d^-1），and a metformin group （0.065 g·kg^-1·d^-1），with 10 mice in each group. Ten FVB mice were assigned to the control group. The mice in the groups with drug intervention were continuously administered correspondingly for 28 days. After administration，the mice were sacrificed，followed by the oral glucose tolerance test （OGTT） and FBG detection. Serum very low-density lipoprotein（VLDL）content was determined by semi-quantitative enzyme-linked immunosorbent assay （ELISA）. Four indexes related to blood lipid were determined by the biochemistry analyzer. Liver tissues were subjected to pathological examination by hematoxylin-eosin（HE）staining. Western blot was used to detect the protein expression of PI3K，Akt，phosphorylated（p）-PI3K，p-Akt，forkhead box protein O1 （FoxO1），insulin receptor（InsR），and insulin receptor substrate-2（IRS-2） in liver tissues of mice. Real-time polymerase chain reaction（Real-time PCR） was used to detect the mRNA expression of PI3K，Akt，FoxO1，InsR，and IRS-2 in liver tissues of mice. ResultCompared with the control group，the model group showed poor general conditions，abnormal glucose tolerance （P<0.05），increased FBG （P<0.01），abnormal blood lipid metabolism，increased serum total cholesterol （TC），triglyceride（TG），low-density lipoprotein cholesterol（LDL-C），and VLDL （P<0.05），decreased level of high-density lipoprotein cholesterol（HDL-C）（P<0.05），fatty degeneration and obvious pathological changes of liver cells，reduced protein expression of PI3K，Akt，p-PI3K/PI3K，p-Akt/Akt，IRS-2，and InsR in liver tissues（P<0.05），increased protein expression of FoxO1（P<0.05），decreased mRNA expression of PI3K，Akt，IRS-2，and InsR in liver tissues （P<0.05），and increased FoxO1 mRNA expression（P<0.05）. Compared with the model group，the BHRS group showed improved general conditions and glucose and lipid metabolism （P<0.05），improved pathological state of liver cells，increased protein expression of PI3K，Akt，p-PI3K/PI3K，p-Akt/Akt，IRS-2，and InsR in liver tissues（P<0.05），decreased protein expression of FoxO1（P<0.05），increased mRNA expression of PI3K，Akt，IRS-2，and InsR in liver tissues （P<0.05），and reduced FoxO1 mRNA expression（P<0.05）. ConclusionBHRS can effectively reduce blood glucose，regulate blood lipid metabolism，and improve the pathological state of the liver in MKR diabetic mice，and its mechanism of action may be related to the regulation of the activity of molecules on the PI3K/Akt signaling pathway.

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult ; Asian Continental Ancestry Group* ; China ; Comorbidity ; Developed Countries ; Developing Countries ; Diagnosis* ; Epidemiologic Studies ; Epidemiology ; Global Health ; Humans ; Hypersensitivity* ; Prevalence ; Rhinitis, Allergic*

OBJECTIVETo screen the HIV-1 entry inhibitors targeting HIV-1 gp120 from the IBS natural product database by virtual screening based on the binding mode of the neutralizing antibody VRC01 with HIV-1 gp120 and investigate the anti-viral activities of the inhibitors and their action mechanisms.

METHODSThe binding interaction of the candidate molecules binding gp120 and changes of the binding free energy were analyzed by MM-PBSA calculation. The anti-HIV-1 activities of the tested compounds were detected by HIV-1 pseudotyped virus, laboratory-adapted HIV-1 and a cell-cell fusion assay. The cytotoxicity of the studied molecules was examined by XTT colorimetric assay. The mechanisms of the anti-viral activities of the candidate molecules were analyzed using enzyme-linked immunosorbent assay.

RESULTSA total of 19 molecules with distinct reduction of the binding free energy after binding with gp120 were screened from 40000 molecules. Among them, NC-2 showed anti-HIV-1 activities against HIV-1 pseudotyped virus and laboratory-adapted HIV-1, and was capable of blocking HIV-1 envelope-mediated cell-cell fusion. The IC50 of NC-2 for inhibiting HIV-1IIIB and pseudotyped HIV-1JRFL infection were 1.95∓0.44 µmol/L and 10.58∓0.13 µmol/L, respectively. The results of ELISA suggested that NC-2 could inhibit the binding of HIV-1 gp120 to CD4 without blocking the formation of gp41 six-helix bundle in vitro.

CONCLUSIONThis computer-based virtual screening method can be used to screen HIV-1 entry inhibitors targeting gp120. Using this virtual screening approach combined with anti-viral activity screening, we obtained a potent HIV-1 entry inhibitor NC-2 with novel structure.

Anti-HIV Agents ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Neutralizing ; pharmacology ; Binding Sites ; Cell Fusion ; Cell Line ; Drug Discovery ; Drug Evaluation, Preclinical ; HIV Antibodies ; pharmacology ; HIV Envelope Protein gp120 ; antagonists & inhibitors ; HIV-1 ; drug effects ; Humans ; Microbial Sensitivity Tests

Objective To investigate the nutritional risks, prevalence of undernutrition, and nutritional interventions among inpatients in departments of nephrology in some hospitals in Guangzhou, with an attempt to provide evidences for the nutritional support of patients with kidney diseases. Methods Totally 378 adult patients in departments of nephrology in Guangzhou were enrolled in this study by fix-point consecutive sampling. Nutritional Risk Screening 2002 (NRS 2002) was applied for nutritional risk assessment. Nutrition risk was defined by NRS score ≥3 and undernutrition by BMI ＜ 18.5 kg/m2 or serum albumin ＜ 30 g/L. Nutritional interventions were also evaluated in all patients. The relationship between nutritional risk and nutritional support was analyzed. Results The overall prevalence of undernutrition was 21.7% and the nutritional risk was 41.3%. They were especially high among patients with chronic kidney dysfunction (24. 3% and 60. 7% , respectively). The nutritional risk was 42. 3% in patients accompanied with diabetes (P＞0. 05). Of these 378 patients, 102 (27.0%) received nutritional interventions, in which the nutritional support rate was 50. 0% (78/156) for patients with nutritional risks and 10. 8% (24/222) for those without nutritional risks. Conclusions The nutritional risks and prevalence of undernutrition are high among inpatients in the departments of nephrology in hospitals in Guangzhou. Proper application of nutritional interventions remains a concern. Evidence-based guidelines are required to improve this situation.