A high-throughput three-dimensional (3D) hepatocyte culture model is constructed in this study. It is capable of replicating the 3D in vivo environment and offers the advantages of high throughput, enhanced reproducibility, low cost, and simplified operation, rendering a valuable tool for hepatotoxicity screening of traditional Chinese medicine (TCM). First, we constructed the 3D high throughput liver chip model using collagen hydrogel. The precision and its difference with traditional cell culture plates were assessed using acetaminophen, Tripterygium hypoglaucum, and Qili San as the references. The feasibility of this model was also investigated by comparing the hepatotoxicity among four batches of T. hypoglaucum and Qili San. The methodology verification shows that the 3D hepatocyte model is better than traditional two-dimensional (2D) cell culture model in precision and feasibility. Subsequently, we compared the hepatotoxicity of different samples over a period of three or seven days and found that the hepatotoxicity of TCM extracts increased over time. Finally, we compared the hepatotoxicity of T. hypoglaucum and its compound formula Kunxian capsule under equivalent concentration. The results showed that the compound Kunxian capsule could significantly reduce hepatotoxicity of T. hypoglaucum. Generally, we constructed a high-throughput, robust 3D liver-on-a-chip model with the potential of rapid assessing TCM-induced liver toxicity.

Objective To observe the therapeutic efficacy of balance cupping plusYang He decoction in treating granulomatous mastitis (GM).Method Eighty GM patients were randomized into a treatment group and a control group according to the last digit of registered number, odd or even, 40 cases each. The control group was treated withYang He decoction; the treatment group was intervened by balance cupping in addition to the treatment given to the control group. They were treated for 6 months in total. The size of lump, swollen size and infection size by the ultrasound B-scanning were observed before and after the treatment, and the clinical efficacies were compared between the two groups.Result The total effective rate was 82.5％ in the treatment group versus 60.0％ in the control group, and the between-group difference was statistically significant (P<0.05). There was a significant difference in comparing the clinical efficacy between the two groups (P<0.05). The size of lump, swollen area and infection size by the ultrasound B-scanning were significantly changed in the two groups after the treatment (P<0.05); the between-group comparisons showed significant differences after the treatment (P<0.05).Conclusion Balance cupping plusYang He decoction can produce a certain efficacy in treating GM.

This study was aimed to investigate the effect of D-limonene on K562 leukemia cells and its mechanism. Inhibitory effect of D-limonene on proliferation of K562 leukemia cells was assayed by MTT method and cell apoptosis was detected by flow cytometry and DNA agarose gel electrophoresis, the morphologic change of K562 cells was observed by microscopy. The results showed that when K562 cells were treated with 0.125 - 1.0 mmol/L of D-limonene for 48 hours, the proliferation of K562 cells was obviously inhibited in dose-dependent manner. Typical morphological changes and the typical DNA ladder on agarose gel electrophoresis for analysis of cellular apoptosis were significantly appeared in D-limonene treated K562 cells. Simultaneously, the sub-G1 peak was found in FCM analysis. It is concluded that the D-limonene can inhibit proliferation of K562 cells in dose-dependent manner, cause cell detained at G1 phase and induce apoptosis of K562 cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclohexenes ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; K562 Cells ; Terpenes ; pharmacology

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Child ; Colonic Neoplasms ; pathology ; Cystadenocarcinoma, Mucinous ; metabolism ; secondary ; Cystadenocarcinoma, Serous ; metabolism ; secondary ; Female ; Humans ; Macrophage Migration-Inhibitory Factors ; metabolism ; Microvessels ; pathology ; Middle Aged ; Neoplasm Recurrence, Local ; Ovarian Neoplasms ; metabolism ; pathology ; secondary ; Stomach Neoplasms ; pathology ; Tumor Suppressor Protein p53 ; metabolism ; Young Adult

BACKGROUNDCyanotic patients have potential growth retardation and malnutrition due to hypoxemia and other reasons. Ghrelin is a novel endogenous growth hormone secretagogue that has effects on growth and cardiovascular activities. The aim of this study was to evaluate the plasma level and myocardial expression of ghrelin and insulin-like growth factor-1 (IGF-1) using an immature piglet model of chronic cyanotic congenital heart defects with decreased pulmonary blood flow.

METHODSTwelve weanling Chinese piglets underwent procedures of main pulmonary artery-left atrium shunt with pulmonary artery banding or sham operation as control. Four weeks later, hemodynamic parameters were measured. Enzyme-linked immunosorbent assay for plasma ghrelin and IGF-1 level measurement were performed. Ventricular ghrelin and IGF-1 mRNA expressions were measured by quantitative real-time polymerase chain reaction.

RESULTSFour weeks after surgical procedure, the cyanotic model produced lower arterial oxygen tension ((68.73 ± 15.09) mmHg), arterial oxygen saturation ((82.35 ± 8.63)%), and higher arterial carbon dioxide tension ((51.83 ± 6.12) mmHg), hematocrit ((42.67 ± 3.83)%) and hemoglobin concentration ((138.17 ± 16.73) g/L) than the control piglets ((194.08 ± 98.79) mmHg, (96.43 ± 7.91)%, (36.9 ± 4.73) mmHg, (31.17 ± 3.71)%, (109.83 ± 13.75) g/L) (all P < 0.05). Plasma ghrelin level was significantly higher in the cyanotic model group in comparison to the control (P = 0.004), and the plasma IGF-1 level was significantly lower than control (P = 0.030). Compared with control animals, the expression of ghrelin mRNAs in the ventricular myocardium was significantly decreased in the cyanotic model group (P = 0.000), and the expression of IGF-1 mRNAs was elevated (P = 0.001).

CONCLUSIONSChronic cyanotic congenital heart defects model was successfully established. Plasma ghrelin level and myocardial IGF-1 mRNA expression were significantly up-regulated, while plasma IGF-1 level and myocardial ghrelin mRNA expression were down-regulated in the chronic cyanotic immature piglets. The ghrelin system may be an important part of the network regulating cardiac performance.

Animals ; Cyanosis ; blood ; metabolism ; physiopathology ; Female ; Ghrelin ; blood ; metabolism ; Heart Defects, Congenital ; blood ; metabolism ; physiopathology ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Pulmonary Circulation ; physiology ; Swine

BACKGROUNDRhoA/Rho kinase (ROCK) pathway is involved in pulmonary arterial hypertension (PAH) and pulmonary artery smooth muscle cell (PASMC) proliferation. Inhibition of ROCK has been proposed as a treatment for PAH. But the mechanism of RhoA/ROCK pathway and its downstream signaling in proliferation of human PASMCs is unclear. We investigated the effect of fasudil, a selective ROCK inhibitor, on platelet-derived growth factor (PDGF) induced human PASMC proliferation, and the possible association between RhoA/ROCK and extracellular signal-regulated kinase (ERK), p27(Kip1).

METHODSHuman PASMCs were cultured with the stimulation of 10 ng/ml PDGF, and different concentrations of fasudil were added before the addition of mitogen. Cell viability and cell cycle were determined with MTT and flow cytometry respectively. ROCK activity, ERK activity and protein expression of proliferating cell nuclear angigen (PCNA) and p27(Kip1) were measured by immunoblotting.

RESULTSBy MTT assay, PDGF significantly increased the OD value that represented human PASMC proliferation, and pretreatment with fasudil significantly reversed this effect in a dose-dependent manner. After PDGF stimulation, the percentage of cells in S phase increased dramatically from 15.6% to 24.3%, while the percentage in G(0)/G(1) phase was reduced from 80.6% to 59%. And pretreatment with fasudil reversed the cell cycle effect of PDGF significantly in a dose-dependent manner. PDGF markedly induced ROCK activity and ERK activity with a peak at 15 minutes, which were significantly inhibited by fasudil. In addition, fasudil significantly inhibited PDGF-induced PCNA expression and fasudil also upregulated p27(Kip1) expression in human PASMCs, which decreased after PDGF stimulation.

CONCLUSIONRhoA/ROCK is vital for PDFG-induced human PASMC proliferation, and fasudil effectively inhibited PDGF-induced human PASMC proliferation by up-regulation of p27(Kip1), which may be associated with inhibition of ERK activity.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; MAP Kinase Signaling System ; physiology ; Muscle, Smooth, Vascular ; cytology ; Platelet-Derived Growth Factor ; pharmacology ; Protein Kinase Inhibitors ; pharmacology ; Pulmonary Artery ; cytology ; Up-Regulation

OBJECTIVETo observe the clinical effect of post-extension pulling massage in treating lumbar disc herniation.

METHODSFrom January 2008 to December 2008, 61 patients with lumbar disc herniation, 34 males and 27 females, ranging in age from 17 to 67 years with an average of 42.6 years, were treated with post-extension pulling massage after continued traction for 30 minutes (on alternate days one time, 3 times as a course of treatment). There was bulging type in 9 cases, hernia type in 22, free type in 30. After a course of treatment, the clinical effects were evaluated according to standard of Macnab, the items included pain, lumbar activity, normal work and life of patients.

RESULTSAll patients were followed up from 1 to 9 months with an average of 4.6 months. After treatment, the symptoms and signs of patients had obviously improved in above aspects. According to standard of Macnab, 48 cases got excellent result, 10 good, 2 fair, 1 poor.

CONCLUSIONThe post-extension pulling massage in treating lumbar disc herniation can obtain satisfactory results, which have localized site of action, small compression for vertebral body and can reduce accidental injury.

Diskectomy ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; pathology ; Lumbosacral Region ; pathology ; Male ; Massage ; methods ; Spine ; Traction ; Treatment Outcome

OBJECTIVETo explore the family aggregation and the role of hereditary factors in the pathogenesis of Kashin-Beck disease (KBD).

METHODSWith a stratified sampling method, the general population of 14 villages of Linyou County were studied, from whom 225 KBD probands were selected using systematic sampling at the rate of (1/2). A total of 304 siblings of the probands were ascertained, and in these sibling pairs, the segregation ratio, heritability in different age groups and weighted mean heritability of the siblings were estimated using the methods of Li-Mantel-Grart and Falconer.

RESULTSThe KBD distribution scope in the KBD families exceeded the scope of binomial distribution (P<0.001), suggesting obvious family aggregation. The prevalence rate in the siblings of the KBD pedigree was 19.41% (59/304), significantly higher than that in the 14 KBD villages [10.90% (1180/10823), chi2=21.62, P<0.001]. The segregation ratio and heritability in the siblings of the KBD pedigrees were 0.061 and 28.61%, respectively.

CONCLUSIONAs a polygenetic inheritance disease, KBD exhibits obvious familial aggregation, and genetic susceptibility accounts for (1/4) of the risk factors for KBD.

Adolescent ; Adult ; Aged ; Child ; China ; epidemiology ; Endemic Diseases ; Family Health ; Female ; Humans ; Male ; Osteoarthritis ; epidemiology ; genetics ; Pedigree ; Prevalence ; Selenium ; deficiency ; Siblings ; Young Adult

BACKGROUNDCyanotic congenital heart defects with decreased pulmonary blood flow due to lung ischemia, hypoxia, and others lead to infant morbidity and mortality more than acyanotic heart disease does. Despite the great effort of medical research, their genetic link and underlying microRNAs molecular mechanisms remain obscure. In this study, we aimed to investigate microRNAs regulation during cyanotic defects in lung of immature piglets.

METHODSCyanotic piglet model was induced by main pulmonary artery-left atrium shunt with distal pulmonary artery banding. Four weeks later, hemodynamic parameters confirmed the development of cyanotic defects and pulmonary lobe RNA was extracted from all animals. We studied the repertoire of porcine lung microRNAs by Solexa deep sequencing technology and quantified highly expressed microRNAs by microarray hybridization. Furthermore, we quantitated selected microRNAs from cyanotic and control piglets by quantitative RT-PCR.

RESULTSAfter surgical procedure 4 weeks later, the cyanotic model produced lower arterial oxygen tension, arterial oxygen saturation, and higher arterial carbon dioxide tension, hematocrit and hemoglobin concentration than controls (all P < 0.05). In 1273 miRNAs expressed in the immature piglets lungs, 2 most abundant microRNAs (miR-370 and miR-320) demonstrated significant difference between cyanotic and control group (all P < 0.05).

CONCLUSIONOur results extended lung microRNA profile in immature piglets and suggested that miR-370 and miR-320 are significantly up-regulated in cyanotic lung tissues.

Animals ; Chronic Disease ; Cyanosis ; genetics ; physiopathology ; Gene Expression Profiling ; Heart Atria ; surgery ; MicroRNAs ; analysis ; physiology ; Pulmonary Artery ; surgery ; Pulmonary Circulation ; Real-Time Polymerase Chain Reaction ; Swine ; Swine, Miniature

OBJECTIVETo analyze the genetic polymorphism of 8 short tandem repeat (STR) loci on human chromosome 2 in Chinese Han population in Shaanxi Province.

METHODSBlood samples anticoagulated with EDTA were collected from 176 unrelated Chinese Han individuals in Shaanxi Province. The DNA was extracted for PCR amplification of the relevant fragments, and the amplified products were analyzed using the ABI 3730 Genetic Analyzer.

RESULTSOn human chromosome 2, the loci D2S112, D2S162, D2S2330, D2S2216, D2S347, D2S259, D2S319 and D2S168 had 7, 11, 9, 8, 9, 9, 8 and 13 alleles, respectively, with 15, 33, 23, 18, 13, 12, 25 and 33 genotypes for the corresponding alleles. The genotype distribution of all the 8 loci met Hardy-Weinberg equilibrium. The heterozygosities for the 8 STR loci were 0.6985, 0.8274, 0.8042, 0.6816, 0.6541, 0.5213, 0.8432 and 0.8091, with polymorphic information content of 0.6911, 0.8199, 0.7891, 0.6809, 0.6388, 0.5187, 0.8372 and 0.8049, respectively.

CONCLUSIONThe 8 loci on chromosome 2 have high heterozygosity and polymorphic information content in Chinese Han population, suggesting their value as useful genetic markers.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, Pair 2 ; genetics ; Genetics, Population ; Genotype ; Heterozygote ; Humans ; Microsatellite Repeats ; Polymorphism, Genetic