2D-DIGE proteomic analysis of changes in estrogen/progesterone-induced rat breast hyperplasia upon treatment with the Mongolian remedy RuXian-I.

RuXian-I has traditionally been used as a remedy for breast hyperplasia in the Inner Mongolia Autonomous Region of China. As a first step toward the investigation of biomarkers associated with RuXian-I treatment, a proteome-wide analysis of rat breast tissue was conducted. First, rat breast hyperplasia was induced by injection of estradiol and progesterone. After treatment with RuXian-I, there is a marked decrease in the hyperplasia, as can be shown by decreases in the nipple diameter and the pathological changes in breast. Subsequently, we used an approach that integrates size-based 2D-DIGE, MALDI-TOF/TOF-MS, and bioinformatics to analyze data from the control group, the model group and the RuXian-I treatment group. Using this approach, seventeen affected proteins were identified. Among these, 15 (including annexin A1, annexin A2, superoxide dismutase [Mn], peroxiredoxin-1, translationally-controlled tumor protein and a B-crystallin) were significantly up-regulated in the model group and down-regulated upon treatment with RuXian-I, and two (Tpil protein and myosin-4) have the opposite change trend. The expression of annexin A1 was confirmed using immunohistochemistry. The expression of superoxide dismutase (SOD) activity was confirmed biochemically. These results indicated that RuXian-I treats rat breast hyperplasia through regulation of cell cycle, immune system, metabolic, signal transduction, etc. The differential expressions of these proteins (annexin A1, superoxide dismutase [Mn], alpha B-crystallins and translationally controlled tumor protein, among others) were associated with occurrence and metastasis of breast cancer. These findings might provide not only far-reaching valuable insights into the mechanism of RuXian-I action, but also leads for prognosis and diagnosis of breast hyperplasia and breast cancer.

[Effect of Drynaria fortunei naringin on the total protein content and ultra-structure of human periodontal ligament cells].

OBJECTIVE: To evaluate the effects of Drynaria fortunei naringin on the total protein content and ultra-structure of human periodontal ligament cells (hPDLCs). METHODS: Through enzyme digestion combined tissue-culture method, primarily culture and identify the human periodontal ligament cells. Coomassie brilliant blue staining was used to detect the total protein content of hPDLCs with the effects of difference concentration of Drynaria fortunei naringin at difference times. Transmission electron microscope was used to study the ultra-structure of hPDLCs with the effects of Drynaria fortunei naringin. RESULTS: In vitro, the addition of Drynaria fortunei naringin at dose of 100, 10, 1, 0.1 mg x L(-1) in cultures resulted in an increase of total protein content at the 3rd, 5th, 7th day, but the maximum response was obtained with 1 mg x L(-1) Drynaria fortunei naringin. There were more rough endoplasmic reticulums, mitochodrias and ribosomes in the experimental group than in the control. CONCLUSION: Drynaria fortunei naringin may stimulate the protein synthesis and metabolism of hPDLCs.

[Effects of Drynaria fortunei naringin on proliferation, alkaline phosphatase activity of human periodontal ligament cells].

OBJECTIVE: To evaluate the biological effects of Drynaria fortunei naringin on the proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLC). METHODS: hPDLC were primarily cultured and identified in vitro through enzyme digestion combined tissue culture method. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to observe the proliferation of hPDLC with the effects of different concentrations of Drynaria fortunei naringin at difference times. The method recommended by International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) was adopted to investigate the effect of Drynaria fortune naringin on the alkaline phosphatase (ALP) activity of hPDLC. At the same time, we used bright blue method to detect the contents of protein in each sample. Then ALP activity in per milligram protein was accounted. RESULTS: Significant proliferative promotion to hPDLC by Drynaria fortunei naringin could be observed at the dose of 10, 1, 0.1 mg x L(-1). Significant ALP activity of hPDLC promotion of Drynaria fortune naringin could be observed at the dose of 100, 10, 1, 0.1 mg x L(-1) and the dose of 1 mg x L(-1) Drynaria fortune naringin had greatest promotion on ALP activity of hPDLC. CONCLUSION: Drynaria fortune naringin might significantly promote the proliferation and increase the ALP activity of hPDLC.

[Effect of an early application of chaiqin chengqi decoction in treating severe acute pancreatitis complicated with acute respiratory distress syndrome].

OBJECTIVE: To investigate the therapeutic effects of an early application of Chaiqin Chengqi Decoction (CQCQD) on severe acute pancreatitis (SAP) complicated with acute respiratory distress syndrome (ARDS). METHODS: Forty patients of SAP-ARDS were equally randomized into the early-treated group (ET) and the late-treated group (LT), CQCQD was administered to them immediately and 3 days later after hospitalization respectively. Baseline materials in the two groups at the entry were insignificantly different (P > 0.05), and the same conventional Western medical therapy were available to them all. The Acute Physiology and Chronic Heath Evaluation II (APACHE I) scores, the incidence and sustained time of complications, the occurrence of infection, requirement of operation shifting on day 7, as well as the duration resided in hospital and mortality in patients were observed and compared. RESULTS: Comparisons of the above-mentioned clinical indexes between groups showed that the APACHE II score was lower (5.1 +/- 2.0 scores vs 9.3 +/- 4.3 scores, P < 0.01); the incidence of shock was lesser (1/20 vs 7/19); the duration of ARDS, renal failure, cardiac insufficiency, hepatic dysfunction, cerebropathy and enteroplegia, as well as the duration in hospital and the requirement of operation shifting were all shorter significantly (P < 0.05) in the ET group than those in the LT group, but no statistical difference (P > 0.05) was shown in terms of the infection incidence and the mortality. CONCLUSION: An early application of CQCQD in the treatment of SAP could shorten the duration of complications and the couse of disease, lower the requirement of operation shifting. But further study with large samples for explore its impact on the infection incidence and the mortality is needed.

[Impact of Chai Qin Cheng Qi Decoction on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis].

OBJECTIVE: To test the change of true choline esterase (TChE) and cholineacetyltransferase (ChAT) and their correlation with interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-alpha), and the impact of Chai Qin Cheng Qi Decoction (CQCQD) on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis (SAP). Methods Thirty SD rats were randomly divided into three groups: control group (pseudo-operated), SAP group and CQCQD-treated group, each with 10 rats. Blood samples were taken six hours after injection of testing agents for biochemical test, which included the test of amylase, TNF-alpha, IL-6, TChE, and ChAT. RESULTS: The rats in SAP group had higher levels of serum IL-6, TNF-alpha and TChE and lower levels of serum ChAT than those in control group (P < 0.05). The serum IL-6 was positively correlated with TChE (r = 0.95, P = 0.000) and negatively correlated with ChAT (r = -0.91, P = 0.000). The TNF-alpha was also positively correlated with TChE (r = 0.93, P = 0.000) and negatively correlated with ChAT (r = -0.95, P = 0.004). The rats in CQCQD-treated group had lower levels of serum IL-6, TNF-alpha and TChE and higher levels of serum ChAT than those in SAP group (P < 0.01). The increase of white blood cell, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase was observed in the control group first and followed by the CQCQD-treated group and SAP group sequentially (P < 0.05). Conclusions Cholinergic anti-inflammatory pathway plays an important role in the pathological changes of SAP in rats. CQCQD can relieve the systemic inflammatory response syndrome and reduce the functional damage of organs through interference on the cholinergic antiinflammatory pathway. More studies are needed to reveal the mechanism of such impact.

[Clinical study on severe acute pancreatitis associated with hypoalbuminemia in early stage].

OBJECTIVE: To investigate the occurring mechanism and clinical characteristics of severe acute pancreatitis (SAP) associated with hypoalbuminemia in early stage and its influence on prognosis of SAP and the preventive and therapeutic management of this disease. METHODS: One hundred and thirty-eight cases diagnosed as SAP complicated by hypoalbuminemia in early stage were accepted in our hospital from August 1, 2003 to December 31, 2004, and they were divided into 2 groups according to the level of plasma albumin: mild hypoalbuminemia (30 to 35 g/L) group and severe hypoalbuminemia (<30 g/L) group. The complications in the early stage, related parameters, and the incidence rate of infection and mortality in the later stage were evaluated respectively. RESULTS: The incidence rates of renal dysfunction, shock, cardiovascular failure and gastrointestinal hemorrhage, the score of acute physiology and chronic health evaluation II (APACHE II ) and the frequencies of pulse and breath in the severe hypoalbuminemia group were all higher than those in the mild hypoalbuminemia group (P<0.05 or P<0.01). The differences of incidence rate of hepatic failure and the scores of Ranson and Balthazar CT between these two groups had no statistical significance (P>0.05). The incidence rate of infection and the mortality in the severe hypoalbuminemia group were higher than those in the mild hypoalbuminemia group (P<0.01) in the later stage of SAP. CONCLUSION: Hypoalbuminemia in the early stage can accelerate the deterioration in pathophysiology of SAP. The lower level of the plasma albumin is in the early stage, the more complications and the higher incidence rate of infection and mortality will be in the later stage. To relieve the extent of systemic inflammatory response syndrome (SIRS) and abundant supplement of albumin, amino acid and lipid in time may be crucial to prevent the occurrence and deterioration of hypoalbuminemia.

[Clinical study of Chaiqin Chengqi Decoction in treating severe acute biliary pancreatitis].

OBJECTIVE: To study the therapeutic effects of Chaiqin Chengqi Decoction (CQCQD) in treating severe acute biliary pancreatitis. METHODS: Ninety patients with severe acute biliary pancreatitis were treated with CQCQD, and they were divided into two groups: early-treated group (54 patients treated with CQCQD within 3 days after the onset of severe acute biliary pancreatitis) and late-treated group (36 patients treated with CQCQD between 3 and 7 days after the onset of severe acute biliary pancreatitis). The complication incidence rate, operation rate, mortality rate and hospitalization period were examined. RESULTS: The incidence rates of encephalopathy, infection and gastrointestinal hemorrhage were lower in the early-treated group than those in the late-treated group (P<0.05). The hospitalization periods of the early- and late-treated groups were (24.9+/-18.4) days and (51.6+/-45.9) days respectively (P<0.05). The general mortality rate was 14.4%. The mortality rate of the early-treated group (7.4%) was significantly lower as compared with that of the late-treated group (25.0%) (P<0.05). The operation rate of the early-treated group (11.1%) was also significantly lower as compared with that of the late-treated group (27.8%) (P<0.05). CONCLUSION: Treating severe acute biliary pancreatitis with CQCQD in early stage may reduce the complication incidence rate, shorten the hospitalization period, and decrease the operation rate and mortality rate.