Objective To identify seven species of Gynostemma BI.,including G.pentaphyllum,G. pentagynum,G.cardiospermum,G.longipes,G.yixingense,G.laxiflorum,and G.guangxiense,by in- ter-simple sequence repeat(ISSR)markers.Methods General DNA was isolated from leaves of the seven species in Gynostemma B1.by CTAB,57 primers constituted by ISSR were tested for PCR and sepharose electrophoresis.Results Fourteen primers amplified polymorphic bands,the amplification patterns of primers UBC-873 and UBC-895 were higher in terms of polymorphic and amplified band ratio.They are used to distinguish all the examined seven species.Conclusion ISSR-PCR Method provides a quick,reli- able molecular marker technique for the identification of different species of Gynostemma B1.

The active ingredients in traditional Chinese medicine have been reported to possess significant pharmacological activity and played an important role in clinical treatments. However, lots of the active ingredients in traditional Chinese medicine suffer from disadvantages such as low solubility, high melting point and low stability that results in low bioavailability and limit its clinical application. Crystal structure plays an important role in improving physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine. This review concludes the research advances of several crystal forms used in the active ingredients in traditional Chinese medicine in terms of polymorph, cocrystal, amorphous/coamorphous and nanocrystal. And the effects of crystal forms on the physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine were reviewed. This research may be useful for the formulation preparation and development of the active ingredients in traditional Chinese medicine.

Objective To investigate the phenotype, frequency of Th17 cells and the association between Th17 cells and viral clearance in patients with H1N1 influenza A. Methods Three groups including 70 confirmed patients with H1N1 influenza A, 30 patients with seasonal influenza as well as 68 healthy subjects as controls were enrolled in this study. The percentages of Th1, Th2, Treg and Th17 lymphocytes in the peripheral blood were determined by intracellular staining and flow cytometry. The levels of interferon-γ (IFN-γ), transforming growth factor-beta (TGF-β),interleukin-6 (IL-6) in plasma and supernatant of the peripheral blood mononuclear cell (PBMC)culture were quantified by enzyme-linked immunosorbent assay (ELISA). Viral load in nasopharyngeal swabs was detected by real time quantitative reverse transcription-polymerase chain reaction (RTPCR). Data were analyzed by one way ANOVA and liner correlation analysis. Results The percentage of Th17 cells in H1N1 influenza A patients was (2. 740±0. 210)%, which the percentage of was significantly decreased compared to healthy subjects (3. 443 ±0. 154)% and seasonal influenza patients (3. 443±0. 277) % (F=4. 242, P＜0. 05); while the percentage of Thl, Th2 and Treg cells were not significantly different among these groups. Moreover, the TGF-β level in plasma of H1N1 influenza A patients was (10±8) ng/mL, which was significantly lower than healthy subjects (43 ±32 ) ng/mL and seasonal influenza patient ( 18 ± 10) ng/mL ( F= 17.72, P＜0.01 ). The TGF-β level in the supernatant of PBMC culture of H1N1 influenza A patients was (782 ± 736) pg/mL, which was significantly lower than healthy subjects (1462±315) pg/mL and seasonal influenza patients (1481 ±348) pg/mL (F=5. 730, P＜0.01). Additionally, the viral clearance period was inversely correlated with the percentage of Th17 cells (r=-0.38, P=0.02). Conclusions The proportion of Th17 cells in patients with H1N1 influenza A is significantly decreased, which is closely correlated with the level of TGF-β. This decrease may results in the delayed viral clearance.

OBJECTIVETo study the role of hypoxia-inducible factor 1 alpha (HIF1alpha) on knee cartilage degeneration,and to explore the effects and mechanisms of Chinese herbal compound Yiqi Huayu prescription on HIF-1alpha gene knockout mice on knee cartilage degeneration.

METHODSThe 4-month and 6-month HIF-1alpha gene knock out mice were obtained by interbreeding, and divided into HIF-1alpha +/+ 4-month mice group,HIF-1alpha -/- 4-month mice group,HIF-1alpha +/+ 6-month mice group and HIF-1alpha -/- 6-month mice group, 3 mice in each group. And then the 2-month-old HIF-1alpha gene knock out mice were randomly divided into Yiqi Huayu prescription group and physiological saline group. There were 6 mice in each group. After 2 months' drug administration, the knee joint of mice was collected, and the Mankin score were evaluated; Safranine-fast green staining, HE Staining, and immunohistochemistry analysis for VEGF, Col X, Col II, MMP-13 and Sox-9 were performed erespectively.

RESULTS(1) Compared to the results in the HIF-1alpha+/+ mice groups, the HIF-1alpha-/- mice developed aging related cartilage loss and bony tissue appearance, cartilage defects increased,and cells reduced. In HIF-1alpha-/-4-month mice and 6-month mice group, the expresion of Col II and Sox9 decreased, and the expression of Col X, MMP-13 and VEGF increased. (2) Compared to the physiological saline group, the ossification and defect of knee joint cartilage reduced of mice in the Yiqi Huayu prescription group, the cartilage cell distribution was more uniform, and the total number of cells increased. The expression of type II collagen and Sox9 protein increased, expression of Col X, MMP-13 and VEGF protein decreased of mice in the Yiqi Huayu prescription group.

CONCLUSIONThe knee cartilage degenerates in the HIF-1alpha cKO mice, and the degeneration increased with age adding. The Yiqi Huayu prescription can delay the degeneration of knee cartilage of HIF-1alpha cKO mice.

Animals ; Cartilage Diseases ; drug therapy ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; deficiency ; genetics ; Joint Diseases ; drug therapy ; genetics ; metabolism ; Knee Joint ; metabolism ; physiopathology ; Male ; Mice ; Mice, Knockout

OBJECTIVETo investigate the clinicopathological characteristics of primary testicular mixed germ cell tumor (MGCT).

METHODSWe retrospectively analyzed the clinicopathological data of 13 cases of primary testicular MGCT and reviewed other relevant literature.

RESULTSMGCT accounted for 24.1% (13/54) of all the testicular germ cell tumors diagnosed in our hospital. The patients ranged in age from 2 to 53 years, averaging at 28.3 years. All were unilateral cases, 6 in the left and 7 in the right testis, with a left/right ratio of 0.86:1. Morphologically, testicular MGCT displayed a variety of subtypes, embryonal carcinoma in 11 cases (84.6%), seminoma in 8 (61.5%), teratoma in 6 (46.2%), choriocarcinoma in 4 (30.8%) and yolk sac tumor in 4 (30.8%). Nine of the cases (69.2%) were composed of two different germ cell histological elements, 3 (23.1%) composed of three, and 1 (7.7%) composed of five.

CONCLUSIONTesticular MGCT is rather rare and most commonly occurs in young men. Its biological behavior, clinical management and prognosis vary with its different histological elements. Therefore accurate pathological diagnosis is essential and immunohistochemistry plays an important role in the diagnosis and differential diagnosis of testicular MGCT.

Adolescent ; Adult ; Child ; Child, Preschool ; Humans ; Male ; Middle Aged ; Neoplasms, Germ Cell and Embryonal ; pathology ; Retrospective Studies ; Testicular Neoplasms ; pathology ; Young Adult

OBJECTIVETo investigate the status of mercury exposure of neonates and their mothers in Zhoushan City, and analyze the related influencing factors.

METHODSFrom Aug to Sep, 2004, 408 neonate-mother pairs were cluster sampled at the Third People's Hospital in Dinghai region, Zhoushan City, and a further investigation for the neurobehavioral development of these children was followed up in a year. The contains of mercury of 14 kinds of fish, 5 pieces of each kind bought in markets of Dinghai region were detected.

RESULTSThe geometric mean of cord-blood Hg was 27.81 nmol/L (5.58 microg/L), while that of hair Hg of mothers was 1246.56 microg/kg. 69.9% of pregnant women had surpassed the RfD (reference dose, 0.1 microg/kg/day) for Hg intake. There was a strong correlation between hair and cord blood Hg levels (r = 0.821). The factors influencing cord-blood Hg levels were fish dinners, dwelling time, and fathers smoking or not, while those of hair Hg were fish dinners and dwelling time. The correlation between fish dinners and the cord blood Hg as well as hair Hg were of significance (r = 0.539, 0.481 respectively), the Hg levels were higher in more fish dinners (H = 119.805, 94.384 respectively). The mean fish Hg level at Dinghai region was low (< 0.1 mg/kg), and no samples were higher than 0.5 mg/L. Two samples of cutlassfish and dried eel contained higher Hg than the others, belonging to a middle level (0.1-0.5 mg/kg).

CONCLUSIONSHg levels in neonates and pregnant women were generally below those considered hazardous, but the safety margin was narrow for some women and supporting efforts should be made to reduce mercury exposure. The main resource of Hg was from eating fish, and it is necessary to investigate the Hg levels in large samples, as to developing the fish advisories for susceptible people.

China ; Environmental Exposure ; analysis ; Environmental Pollutants ; analysis ; Female ; Fetal Blood ; chemistry ; Hair ; chemistry ; Humans ; Infant, Newborn ; Mercury ; analysis ; blood ; Mothers ; Pregnancy ; Seafood ; analysis

OBJECTIVETo investigate the effects of TcpC on human umbilical vascular endothelial cells (HUVECs) and its mechanisms.

METHODSHUVECs were co-cultured with TcpC secreting wild-type E. coli strain CFT073 (TcpC(wt)) or tcpc gene-deleted CFT073 mutant strain (TcpC(mut)) in transwell system,respectively. Apoptosis of HUVECs was analyzed by Annexin-V/PI double staining. Mitochondrial membrane depolarization was detected by JC-1 staining. Expression of apoptosis-related proteins in HUVECs was determined by Western blot.

RESULTSHUVECs showed morphological changes after co-cultured with TcpC(wt) for 24 h: the cells became detached and cell debris increased,and cell number was also decreased when compared to HUVECs co-cultured with TcpC(mut). The apoptosis of HUVEC cells co-cultured with TcpC(wt) for 24 h significantly increased,compared to that of control group and TcpC(mut) group (60.1% 9.7% compared with 9.0% 1.3% and 16.9% 0.4%,respectively, P<0.05); meanwhile the mitochondrial depolarization of HUVECs co-cultured with TcpC(wt) was significantly increased,compared to that in control and TcpC(mut) groups (64.5% 0.9% compared with 14.5% 2.1% and 15.6% 3.3%, respectively,P<0.05). Cleavage of PARP and inhibition of Mcl-1 and XIAP expression were seen in HUVECs co-cultured with TcpC(wt),but not in groups of control and TcpC(mut).

CONCLUSIONTcpC secreted from CFT073 can induce apoptosis of HUVECs through mitochondrial pathway, in which PARP is cleaved and Mcl-1 and XIAP expressions are inhibited.

Apoptosis ; drug effects ; Cells, Cultured ; Escherichia coli ; metabolism ; Escherichia coli Proteins ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; pathology ; Humans ; Membrane Potential, Mitochondrial ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Virulence Factors ; pharmacology ; X-Linked Inhibitor of Apoptosis Protein ; metabolism

OBJECTIVETo investigate the diagnostic value of imageology of giant cell tumour of tendon sheath (GCTS) including X-ray, CT and MRI.

METHODSThirty-five patients with GCTTS confirmed by operation and pathology were retrospectively analyzed. There were 16 males and 19 females. The average age was 39.4 years, ranged from 7 to 66 years. All the patients underwent X-ray examination, 8 patients underwent CT examination, and 16 patients underwent MRI examination.

RESULTSThere were 2 patients in knee joint, 6 patients in ankle joint, 1 patient in capitulum radius, 2 patients in wrist joint, 14 patients in hand and 10 patients in foot. Ten cases were the diffuse form, and 25 cases were the focal form. The X-ray results: the slightly high density soft tissue mass surrounding the bone were shown in 32 cases, 3 cases were normal. The bone erosion were shown in 9 cases, the obvious destruction of bone were shown in 5 cases. CT results: The soft tissue mass and the destruction of bone were shown clearly. MRI results: On T1WI, the signal intensity of GCTTS almost was similar to those of skeletal muscle in 9 cases and was slightly lower than those of skeletal muscle in 7 cases. On T2WI, the signal intensity presented mainly hypointensity with patchy isointensity or hyperintensity signal. A little of fluid was shown in 6 cases.

CONCLUSIONX-ray can demonstrate the lesion and erosion of bone, destruction of bone can clearly be shown on CT. The low intensity signal on MRI T1WI and T2WI is the characteristic appearance of GCTTS. And it can clearly show the lesion range and type of GCTTS.

Adolescent ; Adult ; Aged ; Child ; Diagnosis, Differential ; Female ; Giant Cell Tumors ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Soft Tissue Neoplasms ; diagnosis ; Tendons ; pathology ; Tomography, X-Ray Computed

OBJECTIVETo study the developmental phase on the growth and active compounds in Scutellaria baicalensis.

METHODSeeds of wild plants were collected from Laiwu and sowed in Fangshan (Beijing) and Laiwu (Shandong). Samples of aerial and underground parts were collected in five growth periods of sprouts, seedlings, flowering, seed drop and withered periods respectively. The length of taproot, fresh weight of root, diameter of taproot and the length of stem were determined. The content of active compounds and total flavonoids were determined by HPLC and ultraviolet spectrophotometry respectively. The transcripted level of PAL1, PAL2, PAL3, C4H, 4CL, CHS, GUS and UBGAT were analyzed with RT-PCR.

RESULTThe results showed that the aerial part of S. baicalensis grew quickly before flowering stage, and the underground part grew mostly between the periods of flowering and withered. In the whole growing developmental periods, the content of total flavonoids was not changed significantly, the content of baicalin was increased gradually and the content of baicalein was decreased gradually. Expression level of PAL and 4CL was the highest in withered period, CHS was increased between flowering and seed drop and decreased in withered period.

CONCLUSIONSeedlings and withered periods may be the key phase affecting the growth and active compounds in S. baicalensis.

Acyltransferases ; genetics ; metabolism ; Chromatography, High Pressure Liquid ; Coenzyme A Ligases ; genetics ; metabolism ; Flavonoids ; analysis ; metabolism ; Flowers ; genetics ; growth & development ; metabolism ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Glucuronidase ; genetics ; metabolism ; Glucuronosyltransferase ; genetics ; metabolism ; Phenylalanine Ammonia-Lyase ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plant Stems ; genetics ; growth & development ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; genetics ; growth & development ; metabolism ; Seedlings ; genetics ; growth & development ; metabolism ; Spectrophotometry, Ultraviolet ; Time Factors ; Trans-Cinnamate 4-Monooxygenase ; genetics ; metabolism

OBJECTIVETo study the association between the single nucleotide polymorphisms (SNPs) in the high-temperature requirement A-1 (HTRA1) gene and rheumatoid arthritis (RA) in Chinese Han population.

METHODSFive SNPs in the HTRA1 gene (rs2014307, rs2248799, rs2300433, rs714816 and rs2268356) were genotyped by ABI Snapshot method in Han Chinese cohort composed of 344 patients with RA and 288 healthy controls. The serum rheumatoid factor (RF) and C-reactive protein (CRP) of the patients were determined by endpoint nephelometry method.

RESULTSGenotypes of all the five SNPs in the HTRA1 gene were not significantly different between the RA patients and controls (P> 0.05). Haplotypes generated by these five SNPs did not show significantly difference between the two groups either (P> 0.05). Serum RF levels in the RA patients had no significant difference among the genotypes for four SNPs (rs2014307, rs2248799, rs714816, and rs2268356) in the HTRA1 gene, while RF levels in the RA patients with genotypes AA+AG of the rs2300433 locus were significantly higher than that in genotype GG carriers (P< 0.05). Serum CRP levels in the RA patients had no significant difference among the genotypes for all the five SNPs.

CONCLUSIONAuthor's results suggested that although the five SNPs in the HTRA1 gene were not associated with RA in Chinese Han population, RF levels in the RA patients with genotypes AA and AG in the rs2300433 locus were significantly higher than the GG carriers. The HTRA1 role in RF regulation needs to be further investigated.

Aged ; Arthritis, Rheumatoid ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Haplotypes ; High-Temperature Requirement A Serine Peptidase 1 ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics ; Serine Endopeptidases ; genetics