Permeability is a key factor in the bioavailability of oral drugs. Therefore, in the early stage of drug discovery, accurate and efficient evaluation of drug permeability is essential. The parallel artificial membrane permeability assay (PAMPA) with Caco-2 cells model was used by the industry as early evaluation methods. At present, the Ussing chamber rat model is also widely used. This review summarizes the human data for the in vivo single-pass perfusion technique (Loc-I-Gut) – the gold standard, and then focuses on the basic principles, experimental operation, and efficiency of the three in vitro methods, with correlation to the effective permeability coefficient (P_eff) and fractional absorbed (Fa) in man. We provide recommendations for the use of proper permeability methods at different stages in drug discovery and development.

OBJECTIVETo develop a targeting protein for Xenopus kinesin-like protein 2 (TPX2) C' terminal SBP-3 x Flag-tagged HCT 116 cell model.

METHODSHomologous arms were amplified by polymerase chain reaction (PCR), and then the adeno-associated virus (AAV) -targeting vector of TPX2 was constructed. HCT 116 cells were targeted after the viruses were packaged. Positive cell clones with neomycin resistance gene were obtained by G418 and PCR screening. Finally, the neomycin gene cassette was excised after the targeted clones were infected with adenovirus expressing Cre-recombinase, and the TPX2 C' terminal SBP and 3 x Flag endogenous double-tagged HCT 116 cells were obtained by PCR screening.

RESULTSTwo positive cell clones with neomycin resistance gene were obtained by PCR screening. The positive clones with neomycin resistance gene excised were obtained by Cre adenovirus infection, and the knock-in of SBP-3 x Flag gene was verified by Western blot analysis.

CONCLUSIONThe TPX2 C' terminal SBP-3 x Flag tagged HCT 116 cell model was successfully established.

Cell Cycle Proteins ; genetics ; Colorectal Neoplasms ; genetics ; pathology ; Dependovirus ; genetics ; Gene Targeting ; Genetic Vectors ; HCT116 Cells ; Humans ; Microtubule-Associated Proteins ; genetics ; Nuclear Proteins ; genetics

Objective: To analyze the features and patterns of acupoint selection in acupuncture-moxibustion treatment of chronic atrophic gastritis (CAG) by data mining technique. Methods:Relevant clinical studies published before 25 June, 2017 were searched in databases including PubMed, EMBASE, Cochrane library, Chinese Biomedical Literature Database (CBM), China National Knowledge infrastructure (CNKI), and Wanfang Academic Journal Full-text Database (Wanfang). Results:A total of 122 papers were included, involving 69 points. It was found that the top three points on the frequency list were Zusanli (ST 36), Zhongwan (CV 12) and Weishu (BL 21). The points selected were distributed in 11 meridians, in which the Stomach Meridian of Foot Yangming, Conception Vessel, and Bladder Meridian of Foot Taiyang ranked the top and accounted for 74.0％ of the total frequency. Of the involved specific points, Five Shu-Transmitting points, crossing points and Back-Shu points ranked the top, accounting for 47.1％. The analysis of association pattern has shown that Zusanli (ST 36) and Zhongwan (CV 12) won the highest support rate in the paired groups; Zusanli (ST 36), Weishu (BL 21) and Zhongwan (CV 12) had the highest support rate among the point groups. The Five Shu-Transmitting points and the Lower He-Sea points had the highest support rate among the specific point groups. Conclusion: The data mining results of the studies on acupuncture-moxibustion for CAG are substantially in line with the acupuncture-moxibustion treatment theories in traditional Chinese medicine. The results can reflect the acupoint selection patterns in treatment of CAG and provide reference for acupuncture-moxibustion treatment of CAG in clinic.

OBJECTIVETo investigate the expression of Csk-binding protein (CBP) in esophageal carcinoma and its association with the tumorigenesis and progression of esophageal cancer.

METHODSRT-PCR and Western blotting were employed to determine the expressions of CBP at the mRNA and protein levels in 50 pairs of fresh esophageal carcinoma tissue and the adjacent normal tissues.

RESULTSCBP mRNA and protein expressions in normal tissues were 1.43- and 1.28-fold higher than those in the cancer tissues, respectively (P<0.05). The expressions of CBP mRNA and protein were positively correlated (P=0.015). The decreased expressions of CBP were significantly correlated to lymph node metastasis of esophageal cancer (P<0.05).

CONCLUSIONThe expression of CBP gene is decreased in esophageal carcinoma, which might contribute to the tumorigenesis and progression of this malignancy.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; src-Family Kinases ; genetics ; metabolism

Objective To study the effect of essential oil from Angelica sinensis(EOAS) on neuronal apoptosis induced by ischemia-reperfusion injury.Methods The high differentiated PC12 cells were divided into blank group,model group,control group (10 μmol · L-1 edaravone)and large-,medium-and small-dose expreimental groups(EOAS 25.00,12.50,6.25 μg · mL-1).Except blank group,the remaining groups were treated with sugar-free medium containing 10 mmol · L-1sodium dithionite (Na2S204) for 1 h,reoxygenation for 48 h.The cell proliferation was measured by methyl thiazolyl tetrazolium(MTT) assay.The activity of superoxide dismutase (SOD) and lactate dehydrogenase (LDH),contents of malondialdehyde (MDA) were measured by kit.The apotosis rate and the morphological apoptotic characteristics of cells were observed by flow cytometry and acridine orange/propidium iodide (AO/PI) double staining respectively.The relative activity of caspase-3 were examined with colorimetric assay.The expression of p-ERK1/2 proteins was measured by Western-blot.Results Compared with the blank group,the cell viability in the model group was (67.4 ±0.10)％ with significantly(P ＜0.01).Compared with the model group,the cell viability in the large-dose experimental group and control group were (86.2 ± 0.10)％,(94.5 ± 0.05)％ with significantly (P＜0.05,P ＜ 0.01).Compared with the blank group,the LDH and MDA content in the model group were (912.53±16.71)U · L-1and(9.05 ±0.25)μmol · L-1while SOD level was (12.53 ±0.29)U · mL-1with significantly(all P ＜0.01).Compared with the model group,the LDH and SOD level in the large-dose experimental group were (565.61 ± 11.72) U · L-1 and (12.53 ± 0.29) U · mL-1 with significantly (P ＜ 0.05,P ＜ 0.01) while MDA content in the large-,medium-dose experimental groups were (3.32 ± 0.68),(5.79 ± 0.68) μmol · L-1 with significantly(all P ＜0.01).The absorbance(A) in model group and large-,medium-and small-dose expreimental groups were 0.75 ± 0.06 and 0.10 ± 0.02,0.16 ± 0.03,0.49 ± 0.04.Compared with the model group,the difference had significantly(P ＜0.05,P ＜0.01).Compared with the blank group,the apoptosis rate in the model group was (31.17 ± 2.44)％ with significantly(P ＜ 0.01).Compared with the model group,the apoptosis rate in the large-,medium-dose experimental groups were (4.57 ± 0.32) ％,(5.93 ± 0.81) ％ with significantly (all P ＜ 0.01).Compared with the model group,p-ERK1/2 proteins of large-dose experimental group were up-regulated significantly (P ＜0.01).Conclusion The EOAS could inhibit the apoptosis of neurons after ischemia-reperfusion by activating ERK signaling pathway.

OBJECTIVETo probe into the feasibility of screening anti-HIV compounds by using HIV-1 p24 detection kit made by Hebei Medical University.

METHODSThe sensitivity, reproducibility and efficacy of the Hebei p24 kit were evaluated compared with the commercially available Vironostika HIV-1 Antigen Microelisa System (Biomerieux).

RESULTSHebei p24 kit had high sensitivity and good reproducibility. In vitro screening demonstrated that there was no statistically significant difference (P greater than 0.05) between these two kits in assessing anti-HIV compounds.

CONCLUSIONHebei p24 kit could be used as an easily affordable alternative method for detection of HIV-1 in screening anti-HIV compounds.

Anti-HIV Agents ; isolation & purification ; pharmacology ; Cell Line ; Drug Evaluation, Preclinical ; instrumentation ; methods ; Feasibility Studies ; HIV Core Protein p24 ; analysis ; HIV-1 ; drug effects ; growth & development ; immunology ; Humans ; Reagent Kits, Diagnostic ; standards ; Reproducibility of Results

OBJECTIVETo explore characteristics of the myelin-like bodies in the hepatocytes of patients with Dubin-Johnson syndrome (DJS) complicated with chronic hepatitis B (CHB).

METHODS11 cases of DJS complicated with CHB and 5 cases DJS without CHB were studied clinicopathologically. The hepatocyte ultrastructure was observed with transmission electron microscope and taken photos. The data were compared and analyzed using Fisher's Exact Test.

RESULTSDeposition of myelin-like bodies can be observed in the hepatocytes of DJS patients with CHB but can not in DJS patients without CHB. The morphology of pigment varys. The electron density and volume of pigment in DJS patients with CHB can be classified into five types: brights (2/11,18.2%), reticulation (1/11, 9.1%), punctiform (6/11, 54.5%), abnormity (1/11, 9.1%) and primary type (1/11, 9.1%). The myelin-like bodies in the hepatocytes of patients with DJS are high density and round with membrance (we named it as primary type) (5/5, 100%).

CONCLUSIONSThe myelin-like bodies in the hepatocytes of DJS patients with CHB possess special pleomorphism and may have important diagnostic value.

Adolescent ; Adult ; Female ; Hepatitis B, Chronic ; complications ; pathology ; Hepatocytes ; chemistry ; pathology ; ultrastructure ; Humans ; Jaundice, Chronic Idiopathic ; complications ; pathology ; Male ; Myelin Sheath ; ultrastructure ; Young Adult

Objective To measure the body fat parameters of adult population in Guangxi, and to explore the characteristics of distribution of fat content with age and gender. Methods A total of 1558 male residents and 2132 female residents were selected by random sampling method . The subjects were divided into 8 groups every 10 years, and the fat parameters were measured and recorded by body composition analyzer. The statistical software SPSS 22. 0 analysed data. Results The total of fat mass and fat content of all parts as well as the subcutaneous and limb fat content of all age groups in Guangxi females were higher than that of the male. The fat content of each part of the male increased first and then decreased with age. It was at the age of 30 that the fat parameters of the male peaked with the highest rates of overweight and obesity. Males had more visceral fat and trunk fat than women at 30-40 years old, and showed predominantly centripetal obesity.Total fat content and fat content of all age groups of female showed a rapid rise at the age of 20-50, and reached a peak at 40 years old. After 50 years old, they slowly decreased and gradually stabilized. The main manifestations of young women were subcutaneous and limb fat increase, which was mainly characterized by centripetal obesity after 40-50 years old, when the rates of overweight and obesity were the highest. Conclusion The body fat content of male and female people in Guangxi increases first and then decreases with age. Twenty-thirty years old of Males and 20-50 years old of female are the key time points for the change of body fat content. In addition of the visceral and trunk fat content, the fat content of different age groups and different parts of female is higher in Guangxi than that of males. The visceral and trunk fat content of Guangxi young adults male is higher than that of females.

The flower of Polygonatum cyrtonema has good edible and medicinal values. In this study, four samples of P. cyrtonema flowers from different regions were selected as test materials. The contents, composition and antioxidant activities of lipid-soluble pigments and alcohol-soluble components were determined under different light and temperature conditions, which help to reveal the discoloration reason and the composition variation patterns during storage. The results showed that light and temperature had different effects on the lipid-soluble pigments and alcohol-soluble components in the dried flowers during storage. After storage for 4 weeks, the contents of total chlorophyll, carotenoids, phenols and saponins in the samples exposed to light respectively decreased by 62.62%, 66.4%, 68.7% and 43.4% compared with those in the dark. The decreases in the contents of chlorophyll a, chlorophyll b, lutein, β-carotene and zeaxanthin were 64.64%, 56.74%, 59.2%, 77.7% and 45.4%, respectively. The contents of pigments and components in the samples stored at-20 ℃ were significantly higher than those at room temperature and 4 ℃, indicating that low temperature was conductive to the stability of lipid-soluble pigments and alcohol-soluble components. The samples stored at low temperature and in the dark had the strongest free radical scavenging activity. The results suggest that P. cyrtonema dried flowers should be stored in low temperature environment without light, which can slow down the degradation of internal components. The study provides a theoretical basis for the production, processing and storage of P. cyrtonema flowers.
Antioxidants ; Carotenoids ; Chlorophyll A ; Flowers ; Polygonatum

Objective: To establish a survival prediction model based on the independent prognostic factors of long-term prognosis after laparoscopic liver resection(LLR) for intrahepatic cholangiocarcinoma(ICC). Methods: The clinical and pathological data of 351 consecutive patients with ICC who received radical LLR in 13 Chinese medical centers from August 2010 to May 2021 were collected retrospectively. There were 190 males and 161 females,aged(M(IQR)) 61(14)years(range:23 to 93 years). The total cohort was randomly divided into a training dataset(264 cases) and a validation dataset(87 cases). The patients were followed up by outpatient service or telephone,and the deadline for follow-up was October 2021. Based on the training dataset,the multivariate Cox proportional hazards regression model was used to screen the independent influencing factors of long-term prognosis to construct a Nomogram model. The Nomogram model's discrimination,calibration,and clinical benefit were evaluated through internal and external validation,and an assessment of the overall value of two groups was made through the use of a receiver operating characteristic(ROC) curve. Results: There was no significant difference in clinical and pathological characteristics and long-term survival results between the training and validation datasets(all P>0.05). The multivariate Cox analysis showed that CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis were independent prognostic factors for ICC patients after LLR(all P<0.05). The survival Nomogram was established based on the independent prognostic factors obtained from the above screening. The ROC curve showed that the area under the curve of 1, 3 and 5-year overall survival rates of patients in the training dataset were 0.794(95%CI:0.721 to 0.867),0.728(95%CI:0.618 to 0.839) and 0.799(95%CI:0.670 to 0.928),and those in the validation dataset were 0.787(95%CI:0.660 to 0.915),0.831(95%CI:0.678 to 0.983) and 0.810(95%CI:0.639 to 0.982). Internal and external validation proved that the model exhibited a certain discrimination,calibration,and clinical applicability. Conclusion: The survival Nomogram model based on the independent influencing factors of long-term prognosis after LLR for ICC(including CA19-9,CA125,conversion to laparotomy during laparoscopic surgery,and lymph node metastasis) exhibites a certain differentiation,calibration,and clinical practicability.
Bile Duct Neoplasms/surgery* ; Bile Ducts, Intrahepatic/pathology* ; CA-19-9 Antigen ; Cholangiocarcinoma/diagnosis* ; Female ; Humans ; Laparoscopy ; Lymphatic Metastasis ; Male ; Nomograms ; Prognosis ; Retrospective Studies