Adult human bone marrow-derived mesenchymal stem cells (MSCs) were cultured on microcarriers in spinner flasks and were compared with those in conventional culture in 12-well plates. For the production of adherently growing MSCs, macroporous CultiSpher G gelatin microcarriers were used in concentration of 1 g/L. The cells were seeded in a density of 5 x 10(4) cells/ml in both spinner culture and conventional stationary culture. The result showed that after 7 days of cultivation a maximum viable cell concentration of 5.15 x 10(5) cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675 x 10(5) cells/ml on day 5 in conventional stationary culture. Lactate was produced up to 12.06 mmol/L in spinner culture and up to 13.10 mmol/L in stationary culture, and glucose was consumed up to 7.38 mmol/L and 5.37 mmol/L respectively. The average lactate yield on glucose consumption in spinner culture was only 1.63, lower than that in stationary culture 2.44. This indicated that the energy metabolism in spinner culture was significantly more efficient than that in conventional culture. After spinner culture for 12 days, the MSCs maintain the characteristics of stem cells. It is concluded that the microcarrier culture system is a suitable way to expand the seeding cells for tissue engineering.
Adult ; Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; metabolism ; ultrastructure ; Cell Count ; Cell Culture Techniques ; instrumentation ; methods ; Cell Division ; Cell Survival ; Flow Cytometry ; Glucose ; metabolism ; HLA-DR Antigens ; analysis ; Humans ; Lactates ; metabolism ; Mesoderm ; cytology ; metabolism ; ultrastructure ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Stem Cells ; cytology ; metabolism ; ultrastructure ; Time Factors

In order to efficiently control the quality of the Tibetan medicine Gentianae Szechenyii Flos, the quality standard was established in this study. The tests of water content, total ash and ethanol-soluble extractives of the crude drugs were carried out based on the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC method was established by using reference drug and gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid (7: 1.5: 1: 0.2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on a Ultimate XB-C18 (4.6 mm x 250 mm, 5 μm) column, using methanol-water (0.02% phosphoric acid) (52:48) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is 25 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 10.01-400.32 mg x L(-1) with the regression equation of Y = 1 539.5X - 33.339 (r = 0.999 7), and the average recovery was 99.68% (RSD 1.92%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 19 batches samples were varied in the ranges of 14.48-31.51 mg x g(-1), 11.25% -12.74% and 24.21% - 31.60%, respectively, and total ash was 4.64% - 6.12% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 15.0 mg x g(-1) (1.5%) and 21.0%, respectively; the water and total ash are not more than 13.0% and 6.0%, respectively.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Flowers ; chemistry ; Gentiana ; chemistry ; Medicine, Tibetan Traditional ; Quality Control

Gentianae Urnulae Herba, dried whole herb of Gentiana urnula,is a commonly used Tibetan medicine. However, only the character identification is used as quality control standard officially at present. As a part of project for the Chinese Pharmacopoeia (2015 edition), the quality standard of this species was established in this study. The tests of water content, total ash, acid-insoluble ash and ethanol-soluble extractives of the crude drugs were carried out following the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC identification method was established by using gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid(7:1. 5:1: 0. 2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on an Agilent Zorbax SB-C18 (4.6 mm x 250 mm,5 μm) column, using acetonitrile-water (0.1% phosphoric acid) (26:74) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is at 30 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 0.009 95-0.398 g x L(-1) with the regression equation of Y = 1 467.1X +41.407(r = 0.999 9), and the average recovery was 98. 3% (RSD 2.2%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 15 batches samples were varied in the ranges of 0.175% -1.83%, 8.60% - 9.93% and 29.2% - 35.2%, respectively. Total ash and acid-insoluble ash were 10.2% - 17.2% and 5.26% - 10.8% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 0.80% and 26.0%, respectively; the water, total ash and acid-insoluble ash are not more than 12.0%, 15.0% and 8.0%, respectively.
China ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; standards ; Humans ; Medicine, Tibetan Traditional ; standards ; Plants, Medicinal ; chemistry ; Quality Control

Abstract: Objective To analyze the occupational hazards of enterprises in Pingshan district of Shenzhen in 2017. Methods Occupational hazards were analyzed in 200 enterprises in Pingshan district of Shenzhen City selected using stratified Results random sampling method. A total of 24 industries were involved in the 200 enterprises. The declaration rate of ， occupational hazards was 91.5% and the exposure rate of occupational hazards among workers was 49.2%. The regular monitoring rate of occupational hazard factors in workplaces of the enterprises was 79.5%. There were 129 kinds of occupational ， ， hazard factors of which 19 factors exceeded the national occupational exposure limit accounting for 14.7%. The over standard ， ， ， ， ， ， ， ， rates of noise silica dust cotton dust methanol toluene and other dust were 28.7% 13.6% 11.8% 5.86% 0.5% and ， ， 0.4% respectively. There were 13 kinds of occupational hazard factors in the workplace of metal products industry all of which （ ） exceeded the occupational exposure limit. The exposure rate 56.7% of occupational hazard factors in workers was the highest. Conclusion ， ， The main occupational hazard factors were noise dust and chemical factor and the major occupational hazard industry was metal manufacturing in Pingshan district of Shenzhen City.

To investigate the molecular genetics basis for one para-Bombay phenotype, the red blood cell phenotype of the proband was characterized by standard serological techniques. Exon 6 and 7 of ABO gene, the entire coding region of FUT1 gene and FUT2 gene were amplified by polymerase chain reaction from genomic DNA of the proband respectively. The PCR products were purified by agarose gels and directly sequenced. The PCR-SSP and genescan were performed to confirm the mutations detected by sequencing. The results showed that the proband ABO genotype was A(102)A(102). Two heterozygous mutations of FUT1 gene, an A to G transition at position 682 and AG deletion at position 547-552 were detected in the proband. A682G could cause transition of Met-->Val at amino acid position 228, AG deletion at position 547-552 caused a reading frame shift and a premature stop codon. The FUT2 genotype was heterozygous for a functional allele Se(357) and a weakly functional allele Se(357), 385 (T/T homozygous at position 357 and A/T heterozygous at 385 position). It is concluded that the compound heterozygous mutation--a novel A682G missense mutation and a 547-552 del AG is the molecular mechanism of this para-Bombay phenotype.
ABO Blood-Group System ; genetics ; China ; DNA Mutational Analysis ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Phenotype ; Sequence Deletion

AIMTo study the effects of stress on the cognitive function and physical fitness and its biological mechanisms, provide scientific basis for seeking protective measures to reduce stress-induced damage.

METHODSThe model of restraint stress was adopted in our experiment. Step-down test and exhaustive swimming test were used to measure the learning and memory function and physical fitness in mice, respectively. The contents of GC in plasma, hippocampus LTP, ECG, myocardial ultra-structure, cell apoptosis rate and the level of Hsp70 expression of myocardium in rats were detected.

RESULTSCompared with control group, the impairment of learning and memory function and decline of exercise tolerance were observed in restraint stress group. The elevation in plasma GC levels, ECG abnonrmality, and cell apoptosis rate were also observed under restraint stress. Furthermore, myocardial structure was damaged, and myocardial Hsp70 expression and hippocampus LTP were suppressed in restraint stress group than those in the control.

CONCLUSIONStress may cause the neuro-endocrine dysfunction and homeostasis disorder, and then, lead to the cognitive function and physical fitness damage consequently.

Animals ; Cognition ; physiology ; HSP70 Heat-Shock Proteins ; metabolism ; Hippocampus ; physiology ; Long-Term Potentiation ; physiology ; Male ; Mice ; Myocardium ; metabolism ; Physical Fitness ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Restraint, Physical ; Stress, Physiological ; physiology

OBJECTIVETo study the feasibility and cost-effectiveness of a case-finding program on tuberculosis (TB) in richer rural areas.

METHODSScreening was implemented every three months for a total period of 9 months, in rural areas with high case notification rates. Three villages, each with ten thousand population, were selected to carry out a household screening program. A suspect was defined as who coughed for more than 3 weeks. The suspect was then referred to further diagnosis in county TB dispensary to undergo chest X-ray and sputum test.

RESULTSOf the 86,168 community population screened, 26 TB patients were identified with 7 of them were smear positive. The ratio of effectiveness vs. cost decreased on the second but slightly increased on the third screening program. The direct costs for the 3 screening programs were 6,312,397 and 1637 RMB respectively. Of total direct cost, 5.9% was paid by TB patients, whereas 35.9% was through financing of the county itself.

CONCLUSIONThe community household screening program could achieve higher case detection rate than passive case-finding approach which could be used in richer areas with low case detection rate in China.

China ; Chronic Disease ; Cost-Benefit Analysis ; Cough ; etiology ; Family Characteristics ; Humans ; Mass Screening ; economics ; Radiography, Thoracic ; Rural Health ; Sputum ; microbiology ; Tuberculosis ; complications ; diagnosis

OBJECTIVETo investigate the effects of Kangquan Recipe (KQR) on sex steroids and cell proliferation in an experimental benign prostatic hyperplasia (BPH) model in rats.

METHODSSeventy-two SD rats were randomly divided into six groups: the normal group, the model group, the finasteride group, and the low-, middle-, and high-dose KQR groups, 12 in each group. Except those in the normal group, the rats were injected with testosterone after castration for the establishment of BPH model and then given respectively with normal saline, finasteride, and low-, middle-, and high-dose of KQR for 30 days. The levels of plasma testosterone (T) and estradiol (E(2)) were determined by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression ) of proliferating cell nuclear antigen (PCNA) in prostate tissue was detected by reverse transcription-polymerase chain reaction (RT-PCR) after administration.

RESULTSCompared with the model group, the prostate weight, the plasma T, and the mRNA expression of PCNA were significantly lower, and the plasma E(2) and the ratio of E(2)/T were higher in the three KQR groups (P<0.05 or P<0.01). There was no significant difference in the prostate weight, plasma T and E(2), and ratio of E(2)/T among the finasteride group and the three KQR groups (P>0.05). The mRNA expressions of PCNA were significantly higher in the middle- and low-dose of KQR groups than those in the finasteride group (P<0.05).

CONCLUSIONKQR shows multitarget effects on experimental BPH rats, and the mechanism might be related with regulating the balance of plasma T and E(2) and decreasing the PCNAmRNA expression in prostate tissue to restrain cell proliferation in a dose-dependent manner.

Animals ; Body Weight ; drug effects ; Cell Proliferation ; drug effects ; Cookbooks as Topic ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Gonadal Steroid Hormones ; blood ; metabolism ; Male ; Medicine, Chinese Traditional ; methods ; Organ Size ; drug effects ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Prostate ; drug effects ; pathology ; Prostatic Hyperplasia ; blood ; drug therapy ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of both fermented Cordyceps powder (CS) and prednisone on the Notch2/hes-1 signaling activation in the kidney tubules of rats with acute aristolochic acid nephropathy (AAAN).

METHODSTotally 50 SD rats were randomly divided into 4 groups, i.e., the normal group, the model group, the CS group, the prednisone group, and the CS plus prednisone group, 10 in each group. The AAAN rat model was induced by intragastric administration of pure aristolochic acid A at the daily dose of 100 mg/kg for 3 days. Rats in the CS group were administered with CS at the daily dose of 5.0 g/kg by gastrogavage, while those in the prednisone group were administered with prednisone at the daily dose of 0.5 mg/kg. Rats in the CS plus prednisone group were treated by CS and prednisone. All treatment lasted for 3 successive weeks. Kidney functions [urea nitrogen (BUN) and serum creatinine (SCr)] were detected. The pathological changes of kidneys were observed by Hematoxylin-Eosin staining. The apoptosis of the renal tubular epithelial cells was detected by TUNEL. The protein expressions of Notch2 and Hes-1 in the renal tissue were detected by immunohistochemical assay and Western blot.

RESULTSResults of HE staining showed the structure in the nephridial tissue was regular in rats of the normal group. The renal tubular necrosis occurred in the rats of the model group. The pathological changes of kidneys were obviously improved in the CS group, the prednisone group, and the CS plus prednisone group. Compared with the normal group, levels of BUN and SCr, semi-quantitative score of the tubular interstitial tissue, ratio of apoptotic cells, and expressions of Notch2 and Hes-1 proteins significantly increased in the model group (P < 0.01). Compared with the model group, the aforesaid indices significantly decreased in the 3 treatment groups (P < 0.01). All indices decreased most obviously in the CS plus prednisone group (P < 0.05, P < 0. 01).

CONCLUSIONSNotch2/hes-1 signaling activation might be associated with apoptosis of renal tubular epithelial cells. Both CS and prednisone could play a nephroprotective role for AAAN. But CS plus prednisone could achieve the best effect. Inhabiting the Notch2/hes-1 signaling activation could be its nephroprotective mechanism.

Animals ; Apoptosis ; drug effects ; Aristolochic Acids ; toxicity ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cordyceps ; Female ; Homeodomain Proteins ; metabolism ; Kidney ; metabolism ; Kidney Diseases ; chemically induced ; metabolism ; Kidney Function Tests ; Kidney Tubules ; metabolism ; Male ; Prednisone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptor, Notch2 ; metabolism ; Signal Transduction ; drug effects ; Transcription Factor HES-1

OBJECTIVETo investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC.

METHODSThe expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization.

RESULTThe expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration.

CONCLUSIONThe expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.

Endothelin-1 ; analysis ; genetics ; Fibroblast Growth Factor 2 ; analysis ; genetics ; Hair Follicle ; chemistry ; cytology ; physiology ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Stem Cell Factor ; analysis ; genetics