OBJECTIVE: To investigate biological characteristics of human umbilical cord-derived mesenchymal stem cells, and to explore the possibility of hepatocyte-like cells differentiation.METHODS: The umbilical cord was provided by healthy term birth woman in Tianjin Third Central Hospital. Mesenchymal stem cells were isolated from human umbilical cord by enzyme digestion method. Cells were passaged at 80%-90% confluent. The ninth passage of cells at a density of 5×10~(10)/L were seeded in 12-well culture plate and incubated with DMEM containing hepatocyte growth factor, fibroblast growth factor-4 and oncostatin for 28 days. Cell growth activity was detected by MTT method; cell cycle was detected by flow cytometry; surface immunological marker in MSC was detected by immunocytochemical stain and flow cytometry; specific surface phenotype of hepatocyte was detected by immunocytochemical staining. Function characteristic of hepatocyte was determined by staining for glycogen.RESULTS: MSCs were isolated from human umbilical cord and presented with fibroblastic morphology. 80% of cells were at G_0/G_1 phase with good growth activity and stably passaged over 20 times. These cells were positive for CD29, CD105, and Vimentin, but negative for CD34 and CD31. MSCs were induced to hepatocyte-1 ike cells that were positive for alpha fetoprotein, CK18, CK19 at 1 week and albumin at 3 weeks. At 4 weeks, induced cells were positive for glycogen staining.CONCLUSION: MSCs isolated from human umbilical cord can be cultured in a long periods time in vitro and are able to differentiate into functional hepatocyte-like cells.

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including (+)-licarin A (2), naringenin 4', 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this is the first report of compounds 2, 5 and 6 from Nardostachys genus.
Caffeic Acids ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Lignans ; chemistry ; isolation & purification ; Nardostachys ; chemistry ; Plant Roots ; chemistry ; Rhizome ; chemistry ; Vanillic Acid ; chemistry ; isolation & purification

Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.

Objective To establish a method for determining the contents of polyphyllin Ⅰ and polyphyllin Ⅱ in Kangkeling Mixture. Methods The contents of polyphyllin Ⅰ and Ⅱ were determined by HPLC gradient elution. Poroshell 120 Ec-C18 column (4.6 mm × 150 mm, 4 μm) was used; Acetonitrile-water (A:B) was set as the mobile phase; the flow rate was 1.0 mL/min; the detection wavelength was 210 nm; column temperature was 30 ℃. Results Polyphyllin Ⅰ showed good linear relationship in the range of 1.009–10.09 μg (r=0.999 6), and the average recovery was 97.31% (RSD=2.05%, n=6). Polyphyllin Ⅱ showed good linear relationship in the range of 0.640 5–6.405 μg (r=0.999 8), and the average recovery was 96.41% (RSD=1.67%, n=6). Conclusion The method is simple, with good repeatability and accurate results, which can be used to determine the contents of polyphyllin Ⅰ and Ⅱ in Kangkeling Mixture.

Objective To understand species, population density, and seasonality of rodents in Fangshan District of Beijing and provide the information for prevention and control of rodents and rodent-borne diseases. Methods The rodents were captured by using the trap at-night method in Mukezhai, Xianfenggu, Liushilu, Gushanzhai, Wanjingxiangou.The rodents pieces were identified and the density was calculated. Results Totally 4 665 traps were placed and 268 rodents were captured during 2016-2017.Among the seven identified species, the proportion of Apodemuspeninsulae was the highest (31.0%).The density of rodents was higher in primary forest area (7.29%) and edge area (6.10%) than in living area (2.94%)(P < 0.05).The density of rodents was 6.43% in 2016, and 4.44% in 2017(P < 0.05).The highest density (10.7%) was found in September. Conclusion The rodent species with highest density was Apodemuspeninsulae that can transmit Yersinia pestis.Comprehensive prevention measures should be taken to reduce the rodent density for controlling rodent-borne diseases.

According to the theory of traditional Chinese medicine (TCM), Qi (vital energy) is regarded as a driving force of biological activities in human body, including both nutrient substances and organ functions. Qi-invigorating TCMs are widely used to treat various symptoms and disorders, such as fatigue, obesity, immunosuppression, intestinal flora imbalance, and gastrointestinal diseases, in which Qi is considered to be reduced or depleted. Interestingly, abundant clinical evidences suggest that these disorders are associated with the alternation of intestinal flora, which directly affects disease status. Herein we review the interaction between gut microbiota and Qi-invigorating TCMs under healthy and disease conditions and discuss the mechanisms of action and applications of Qi-invigorating TCMs in enhancing health status through microbial alternation. A better understanding of the role of Qi-invigorating TCMs in modulating microbial composition and the association between intestinal microbiota and diseases would help reveal the clinical consequences of microbiota alteration and explore opportunities to harness this symbiotic relationship to improve public health.
Drugs, Chinese Herbal ; pharmacology ; Gastrointestinal Microbiome ; drug effects ; Humans ; Prebiotics ; Qi

OBJECTIVETo seek a safe, efficient, and cost-effective technique for local thermo-ablation of hepatic cancer.

METHODSThe livers from 16 healthy rabbits were thermocoagulated by diode-laser with scanner fiber tip, 6 w for 10 mins. At the same time, the temperatures were measured at 0, 5 and 10 mm from laser tip. The pre-thermocoagulative liver function was compared with that of 7 days post-thermocoagulation. The pathologic changes were also observed 1 month after laser thermocoagulation.

RESULTSAll the rabbits survived and hepatic tissue temperatures at 0, 5, 10 mm from laser tip reached 96.39 degrees C +/- 3.97 degrees C, 60.79 degrees C +/- 6.21 degrees C, 46.10 degrees C +/- 4.58 degrees C respectively after 10 minutes of thermocoagulation. There were no significant differences in liver function parameters between rabbits of pre-laser thermocoagulation and of post-laser thermocoagulation. Thermocoagulated necrosis of liver tissue with surrounding fibrosis in a diameter of 26.0 mm was formed. Light microscopy revealed coagulative necrosis in the center of the coagulated area without surviving hepatic cells.

CONCLUSIONThe hepatic tissue can be coagulated safely and effectively by diode-laser with scanner fibertip, and such a technique may provide a new method for the treatment of hepatic carcinoma.

Animals ; Female ; Laser Coagulation ; methods ; Liver Neoplasms ; pathology ; surgery ; Male ; Rabbits

OBJECTIVETo determine the effect of three Jianpi Buqi recipes on intestinal microflora of Pi-qi-deficiency rat with molecular biology methods and attempt to clarify the mechanisms of therapy.

METHODRat model of Pi-qi-deficiency was induced by exercise fatigue simultaneously diet incontinence and then administrated with the decoctions of Si-jun-zi-tang (including crude drug) (10.8, 3.6, 1.2 g x kg(-1)), Li-zhong-tang (16.2, 5.4, 1.8 g x kg(-1)) and Buzhong Yiqi-tang (14.4, 4.8, 1.6 g x kg(-1)) recipes which were in common use of clinic of traditional Chinese medicine (TCM). With application of ERIC-PCR (enterobacterial repetitive intergenic consensus-PCR) fingerprint analysis, the efficacy on Pi-qi-deficiency and perturbation of intestinal mircoflora of these recipes were examined and evaluated.

RESULTRemarkable differences of ERIC-PCR profile were found between Pi-qi-deficiency state rats and normal rats. The Shannon's Index (H') was increased when treated with all doses of three Jianpibuqi recipes decoctions and Sorenson's pairwise similarity coefficient (Cs) was also increased significantly after treatment.

CONCLUSIONERIC-PCR fingerprint can be utilized as an initial screening of changes of the composition of bacterial communities that are associated with the development of intestinal disease. It can also be used to build a better understanding of Pi-qi-deficiency and to investigate the pharmacodynamic effect of TCM on Pi-qi-deficiency. Three Jianpi Buqi recipes are demonstrated to produce pronounced recovery activities on intestinal microflora of Pi-qi-deficient rats, which may be considered one of mechanisms of their performances.

Animals ; Bacteria ; drug effects ; genetics ; growth & development ; Drugs, Chinese Herbal ; pharmacology ; Feces ; microbiology ; Female ; Intestines ; microbiology ; Male ; Polymerase Chain Reaction ; Rats ; Rats, Wistar

This study was aimed to analyze the correlation of JAK2V617F mutation burden with clinical features in patients with polycythemia vera (PV) and essential thrombocythemia (ET), The JAK2V617F mutation ratios in 47 PV samples and 43 ET samples were detected by real-time PCR. The correlation of mutation allele ratio in PV and ET samples with clinical features (hemoglobin, hematocrit, white blood cell count and platelet count) was analyzed. The results showed that the JAK2V617F mutation burden was higher in PV (0.441 +/- 0.270) than that in ET (0.209 +/- 0.192). The JAK2V617F mutation burden was positively correlated with levels of hemoglobin (PV: R = 0.518, p < 0.001; ET: R = 0.528, p = 0.005), hematocrit (PV: R = 0.510, p < 0.001; ET: R = 0.524, p = 0.005) and leukocyte (PV: R = 0.584, p = < 0.001; ET: R = 0.471, p = 0.013) in PV and ET samples. The higher JAK2V617F mutation burden was negatively correlated with levels of platelet count in PV samples (R = -0.354, p = 0.020), but there was no correlation between the JAK2V617F mutation burden and platelet count in ET samples (R = 0.233, p = 0.242). It is concluded that the higher JAK2V617F mutation burden is related with higher hemoglobin, hematocrit and leukocyte count in both PV and ET samples. The higher JAK2V617F mutation burden is correlated with lower platelet count in PV samples, but there is no correlation between JAK2V617F mutation burden and platelet count in ET samples.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Hemoglobins ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Mutation ; Polycythemia Vera ; genetics ; Polymerase Chain Reaction ; Thrombocythemia, Essential ; genetics ; Young Adult