A HPLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to establish a determination method for drug concentrations of costunolide (Co) and dehydrocostuslactone (De) in blood samples in the positive ion mode, with diazepam as the internal standard substance, in order to study the pharmacokinetic process of sesquiterpene lactones costunolide and dehydrocostuslactone after the oral administration of Weichang'an pills, and provide an theoretical basis for further studies on the substance basis for the anti-diarrhea effect of Weichang'an pills. In the blood samples, Co and De showed a good linearity within concentration ranges 0.700 0-769.7, 2.510-956.0 μg x L(-1), respectively. The results of precision, stability and recovery experiences proved the stability and reliability of the plasma concentration determination method. After the oral administration, the concentrations of Co and De in plasma increased with the increase in dose, with T(max) between 10.65-12.98 h, indicating a long time to reach peak plasma concentrations; C(max) of costunolide and dehydrocostuslactone ranged between 3.750-5.450,15.34-44.52 μg x L(-1), respectively. The in vivo adsorption of Co and De conformed to the one-compartment model, with a longer time to attain the peak plasma concentrations. These results provided an experimental basis for revealing the active substance basis and clinical medication of Weichang'an pills.
Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Lactones ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Wistar ; Sesquiterpenes ; administration & dosage ; blood ; pharmacokinetics ; Tablets ; administration & dosage ; chemistry ; pharmacokinetics

BACKGROUND:Angiotensin II receptor antagonists have been found to exerct a stronger protective effect on bone than angiotensin converting enzyme inhibitors. OBJECTIVE:To investigate the effect of different concentrations of irbesartan (angiotensin II receptor antagonist) on the differentiation and mineralization of mouse preosteoblasts. METHODS:Mouse preosteoblast cel lines MC3T3-E1 in logarithmic phase were selected and cultured in the osteogenic induction medium containing 0 (control group), 0.001, 0.01, 0.1 mmol/L irbesartan, respectively. Ten days later, the cel differentiation was observed by alkaline phosphatase staining. The mineralization was observed by alizarin red staining after 21 days of culture. mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 in osteoblasts were detected by real-time PCR at 1, 4, 7, 14 and 21 days of culture. RESULTS AND CONCLUSION:The activity of alkaline phosphatase in al the irbesartan groups (0, 0.001, 0.01, 0.1) was higher than that in the control group (P<0.05), which was the most obvious in 0.01 mmol/L. The number and area of calcium nodules in each irbesartan group were significantly higher than those in the control group (P<0.05), especial y in 0.01 mmol/L. Compared with the control group, 0.01 mmol/L irbesartan significantly upregulated the mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 (P<0.05). These results suggest that 0.01 mmol/L irbesartan significantly promotes the differentiation and mineralization of osteoblasts.

Objective To study the influence of low-iodine diet on the expression of homeobox gene Nkx-6.1 and Nkx-6.2 in rat cerebrum tissue, and to explore the possible molecular mechanism of cerebrum development retardation caused by low-iodine. Methods Twenty female Wistar rats were randomly equally divided into two groups: low-iodine group and control group, both fed with low-iodine diet as low as 13.66 μg/kg determinated by spectrophotometry in Tianjin Institute of Endocrinology and the former with deionized water, the later 200 μg/L potassium iodate. Thyroid hormone level was detected using chemiluminescence immunoassay 3 months later and they were mated with male rats normally fed. Rats of 16-day pregnancy, new-born and 20th days old were detected the content of Nkx-6.1 and Nkx-6.2 mRNA in the cerebrum tissue by real-time fluorescence quantitative PCR 0.61), (3.28±0.80)pmol/L] were lower than the control group[(1.04±0.06), (39.42±14.68)nmol/L, (4.83±0.33), day pregnancy, new-born and 20th days old of control group was (1.90±0.23)×10-3,(1.86±0.40)×10-4, (1.11± 0.27)×10-4(F=827.58, P<0.01), Nkx-6.1 mRNA expression level gradually decreased along with aging(all P<0.05). The intra-group difference was significant (F=297.25, P<0.01) and the Nkxr.1 mRNA expression level during 16 days of pregnancy was the highest(P<0.01). It was higher in the control group than in the low-iodine group during 16 days of pregnancy (t=10.14, P<0.01) as well as in the low-iodine group than in the in 16-day pregnancy, new-born and 20th days old of control group was respectively(1.03±0.19)×10-2, (1.33± 0.10)×10-3, (8.79±0,87)×10-3, and that of low-iodine group was (0.31±0.03)×10-2, (1.53±0.13)×10-3, (7.51±0.86)×10-2. The intra-group difference was significant(F=1293.02,1065.83, all P<0.01). Nkx-6.2 expression level during 20th days old was the highest(P<0.01) and that of newborn was the lowest(P<0.01). The Nkx6.2 mRNA expression level in control group were higher than the low-iodine group during 16-day pregnancy and 20th days old(t=14.35, 4.05, all P<0.01). It was higher in the low-iodine group than in the control group during newboru(t=4.78, P<0.01). Conclusions The difference in the expression of Nkx-6.1 and Nkx-62 is highly related to the brain development retardation caused by low-iodine.

Objective To study the characteristics of mismatch negativity(MMN) in patients with depression.Methods MMN was recorded from 31 patients with depression and 28 age-matched controls. Results Compared with the controls, the patients with depression had prolonged latency ([189?22] ms vs [206?26] ms, P

Objective:To explore the influence of low-iodine diet on the expression of homeobox gene NKX-2.2 in rat cerebrum tissue,and to explain the possible molecular mechanism of cerebrum development retardation caused by low-iodine.Methods:Female Wistar rats were randomly divided into two groups:Low-iodine group and control group,both fed with low-iodine feed,given the deionized water and KIO3 solution respectively,they were drawn from the 16-day pregnancy,new-born and 20th days old low-iodine and normal age offspring after three months,and detect the content of NKX-2.2 mRNA in the cerebrum tissue by Real-time fluorescence quantitative PCR techniques.Results:The thyroid hormone levels of low-iodine group in serum were significantly lower than the control group(P

OBJECTIVETo evaluate the risk factors of lateral lymph node metastasis in advanced lower rectal cancer and its correlation with local recurrence and prognosis.

METHODSData from 96 consecutive patients with advanced lower rectal cancer underwent curative surgery with lateral dissection were retrospectively analyzed. The correlations of lateral lymph node metastasis with clinicopathologic characteristics, local recurrence and prognosis were investigated.

RESULTSLateral lymph node metastasis was observed in 14.6 (14/96) of the cases. In 40 patients with tumor diameter > or = 5 cm, 10 (25.0%) patients were found with lateral lymph node metastasis; while in the other 56 patients, only 4 (7.1%) cases were found with lateral lymph node metastasis (P < 0.05). Lateral lymph node metastasis was more frequent in patients whose tumor infiltrated full range of the intestinal wall (70%) than patients with 3/4, 2/4 and 1/4 intestinal wall was infiltrated (12.0%, 6.7% and 6.3%, respectively) (P < 0.05). Lateral lymph node metastasis rate of poorly differentiated carcinomas was significantly higher than those of moderate and well-differentiated ones (30% vs. 9.1% and 4.5%, P < 0.05). Local recurrence occurred in 18.8% (18 of 96 cases) of patients. Local recurrence in patients with positive lateral lymph node metastasis was 64.3%, while 11.0% in those without lateral lymph node metastasis (P < 0.05). Kaplan-Meier survival analysis showed significant improvements in median survival for patients with negative lateral lymph node metastasis over patients with lateral lymph node metastasis (80.9 +/- 2.1 vs. 38 +/- 6.7 months, log-rank P < 0.05).

CONCLUSIONSTumor diameter, degree of tumor infiltration and histological differentiation are significant risk factors of lateral lymph node metastasis in advanced lower rectal cancer. Lateral lymph node metastasis is an important predictor of local recurrence and prognosis of patients.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Prognosis ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies

OBJECTIVETo investigate the relationship between the helicobacter pylori (HP) infection and the genetic instability of mitochondrial DNA (mtDNA) in human gastric adenocarcinoma epithelial cells (AGS).

METHODSAfter treated with extracts of HP11638 (CagA+, VacA+) or Hp11638 mutant strain (CagA+, VacA-), AGS cells were collected, and mitochondrial DNA was extracted and Cox-I, Cox-II, Cox-III, ATPase6, ATPase8 and Cytb genes and the D-Loop region were amplified by PCR and then sequenced.

RESULTSThe mutation rates of the mtDNA in AGS cells were correlated with the extracts of the two HP strains in a concentration- and time-dependent manner. But the mtDNA mutation rate in AGS cells treated with the HP11638 extract was higher than that treated with the Hp11638 mutant extract. Total of 616 mutations in D-Loop region were detected, including 489 point mutations, 81 insertions and 46 deletions. Among them, 70.9% (437/616) belonged to GC to AT and AT to GC transition. Seventeen out of 20 (85%) AGS cells treated with extract of HP had mutations in 303PolyC, 16184PolyC and 514CA regions of mtDNA D-Loop. No mutation was detected in Cox-I, Cox-II, Cox-III, ATPase6 and ATPase8 genes, three point mutations were found in the Cytb gene.

CONCLUSIONHP can cause the accumulation of mutations in mtDNA, in particular, in the D-Loop region, and the VacA participated in the process.

Antigens, Bacterial ; pharmacology ; Base Sequence ; DNA, Mitochondrial ; drug effects ; genetics ; Endothelial Cells ; drug effects ; pathology ; Helicobacter Infections ; complications ; Helicobacter pylori ; chemistry ; Humans ; Mutation ; Stomach ; pathology

OBJECTIVETo study the relationship between microbial gene 16SrRNA and intrauterine infection.

METHODSThirty cases of single preterm birth were enrolled, including 16 cases due to premature rupture of membranes (PROM) (rupture time>18 hrs), 6 cases due to spontaneous preterm birth and 8 cases due to iatrogenic preterm birth. Ten cases of single term birth were used as the control group. Fetal membrane and placenta samples were obtained. Amniotic fluid, blood from cord or newborn babies as well as gastric fluid and tracheal secretions from infants with mechanical ventilation were also obtained. The histological features of placenta and fetal membranes were observed. Polymerase chain reaction (PCR) was used to detect the presence of microbial 16SrRNA and ureaplasma urealyticum (UU) in placenta, fetal membranes and other samples.

RESULTSTwenty-one (70%) cases were diagnosed as chorioamnionitis, characterized by neutrophil infiltration in fetal membrane and placenta tissues, especially in fetal membranes. Chorioamnionitis was most frequent in babies whose gestational age less than 32 weeks or birth weight lower than 1 500 g. Positive 16SrRNA gene was found in 12 cases, and positive UU gene in 10 cases in the preterm birth group. Neither 16SrRNA nor UU gene was detected in the control group. The PROM preterm babies developed more frequent infection than the babies premature born due to other causes, but there were no statistically significant differences in the incidence of infection.

CONCLUSIONSChorioamnionitis may be the major cause of PROM and premature birth. The detection of microbial genes is valuable in identification of intrauterine infection.

Chorioamnionitis ; diagnosis ; Female ; Fetal Membranes, Premature Rupture ; etiology ; Humans ; Infant, Newborn ; Infant, Premature ; Placenta ; microbiology ; pathology ; Pregnancy ; RNA, Ribosomal, 16S ; genetics ; Ureaplasma urealyticum ; genetics ; isolation & purification

Objective To investigate the effects of panax notoginseng saponins (PNS) combined with total flavonoids from epimedium (TFE) on testicular dysfunction in natural aging rats; To discuss its mechanism of action. Methods Thirty 18-month old male SD rats were randomly divided into natural aging group, PNS combined with TFE low and high dose groups, with 10 rats in each group. Another 10 2-month old rats were set as young control group. PNS combined with TFE low and high dose groups were given gastric gavage of 10 mg/kg PNS combined with 10 mg/kg TFE, and 20 mg/kg PNS combined with 20 mg/kg TFE, respectively. Rats in young control group and natural aging group were given saline for 6 d each week, lasting for 4 months. Then, rats were sacrificed, and the testes were obtained to calculate the testicular weight and the testicular index. The testicular tissue morphology was observed by using HE staining. Testicular germ cell apoptosis was detected by using TUNEL method. The levels of Bcl-2, Bax and γ-H2 AX protein expression in testicular tissue were detected by Western blot. Results Compared with natural aging group, low and high dose of PNS combined with TFE significantly elevated the testicular weight and testicular index, improved the histological changes of testicular seminiferous tubule, significantly reduced number of apoptosis of spermatogenic cells in the testis, upregulated the expression of Bcl-2 protein in the testis, downregulated the expression of Bax and γ-H2 AX protein, and decreased the ratio of Bax/Bcl-2. Conclusion PNS combined with TFE can improve testicular dysfunction in natural aging rats by inhibiting apoptosis and DNA damage of germ cells.

Construction of mutant strain is an essential method in pathogenesis researches. The conventional method for Brucella unmarked deletion mutant construction is based on suicide plasmid, but the efficiency is very low. In the present study, we first optimized the electroporation parameters, and then, the cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of the type IV secretion system. This indicated that by using conventional cloning plasmid as suicide plasmid in Brucella, unmarked deletion mutants can be constructed with high efficiency.