Syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid are three main bioactive ingredients in herbs of Saussurea involucrata with various pharmacological properties, while their contents are very low. In this study, the biosynthesis of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid in the cell suspension cultures of S. involucrata were regulated by feeding carbon sources and precursors, which resulted in a great increase of the contents and yields of the above three bioactive ingredients. After 16 days of fermentation, the yields of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid reached 339.0, 225.3, 512.7 mg x L(-1), respectively. Meanwhile, their contents increased up to 67.9, 1.9, 10.6 times of wild medicinal material, respectively. The results provided a solid basis for further studies on application of cell suspension cultures of S. involucrata for large-scale production of bioactive compounds syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid.
Cell Culture Techniques ; Cells, Cultured ; Chlorogenic Acid ; analysis ; metabolism ; Cinnamates ; analysis ; metabolism ; Glucosides ; analysis ; biosynthesis ; Phenylpropionates ; analysis ; Saussurea ; chemistry ; growth & development ; metabolism

Objective:To observe the clinical efficacy of auricular point sticking at different points to relieve the pain in arteriovenous fistula puncture. Methods: A total of 42 patients with arteriovenous fistula were randomized into a Shenmen (TF4) group and an Elbow (SF3) group by the random number table method, with 21 cases in each group. After enrolled into different groups, before the dialysis, patients were given auricular point sticking with Wang Bu Liu Xing ( Semen Vaccariae) seeds at Shenmen (TF4) and Elbow (SF3), respectively. Patients were asked to press the seeds themselves for 2 min each time, four times a day, and an additional 5-15 min before the arteriovenous fistula puncture. Intensive pressing was offered during the puncture, 15-20 presses for each time, and the plasters were changed every 2-3 d. The numerical rating scale (NRS) was used to score the pain level one week before and after auricular point sticking. The NRS score was then compared and analyzed. Results: The intra-group comparison showed that the changes of NRS score in both groups were statistically significant after auricular point sticking (both P＜0.05). After the treatment, there was no significant difference in NRS score between the two groups (P＞0.05). Conclusion: Auricular point sticking at Shenmen (TF4) or Elbow (SF3) can effectively relieve the pain of arteriovenous fistula puncture, and these two points have equivalent analgesic effect.

Objective To investigate the incidence of hearing disorder and analyse the high-risk factors with hearing injury in newborns with hyperbilirubinemia.Methods The newborns with hyperbilirubinemia who admitted to the department of neonate,were received the distortion product otoacoustic emission(DPOAE)test when they recovered from hyperbilirubinemia;those babies who didn′t pass the first test received screening again in 42 days after birth.Those babies who didn′t pass the second test received auditory brain stem response(ABR)test.Results Fifty-eight(33.2%)newborns didn′t pass the first DPOAE test among 235 newborns with hyperbilirubinemia;11(18.9%)infants didn′t pass the second DPOAE test among 58 infants;5 infants failed to pass the ABR test,the ratio of hea-ring disorder in newborns with hyperbilirubinemia was 2.13%;18(9.9%)newborns didn′t pass the first DPOAE test among 182 normal newborns,and those infants all passed the second DPOAE test.Conclusions Hyperbilirubinemia is high-risk population of hearing disorder.The congenital cytomegalovirus infection,neonatal septicemia and hemolytic disease of newborn are the high risk factors responsible for hearing disorder.All high risk newborns should recieve hearing examination regularly.

BACKGROUND: At present, studies have shown that bone marrow mesenchymal stem cells (BMSCs) have self-renewal ability, which can be used as ideal seed cells for repairing tissue and organ damages caused by aging and lesions. OBJECTIVE: To study the changes in the levels of oxidation, inflammatory factors and neurotrophic factors (BDNF) in the brain of aging rats undergoing BMSCs transplantation, and to analyze the mechanism underlying the repair of learning and memory ability in the aging rats. METHODS: A total of 30 clean Sprague-Dawley rats were randomly divided into control group, model group and BMSCs group, 10 rats in each group. Aging models were made in the rats by 3-month subcutaneous injection of D-galactose. After modeling, BMSCs treatment was performed via tail vein injection in the BMSCs group.The injection was performed once a week,for 8 continuous weeks.Morris water maze was used to detect the learning and memory abilities of the rats in each group after the final injection of BMSCs. Superoxide dismutase activity in the brain tissue of rats was detected by xanthine oxidase method. Level of malondialdehyde in the rat brain tissue was detected by thiobarbituric acid method. Total antioxidant capacity of the brain tissue was detected by Fe3+reduction method. Real-time PCR and western blot assay were used to detect the expression of brain-derived neurotrophic factor mRNA and protein in the brain tissue of the aging rat, respectively. RESULTS AND CONCLUSION: Compared with the model group, the BMSCs group exhibited significantly higher activity of superoxide dismutase, stronger total antioxidant capacity, and higher levels of brain-derived neurotrophic factor mRNA and protein (P < 0.05), but the lower malondialdehyde level in the brain (P < 0.05). Compared with the model group, there was less time and higher frequency for passing through the platform in the BMSCs group (P < 0.05). Our findings further indicate that BMSCs can improve the abilities of learning and memory in aging rats, and the underlying mechanism is likely to improve antioxidant capacity and to regulate the level of brain-derived neurotrophic factors.

Objective To establish Ad-hLIF transgenic feeder cells for the expansion of umbilical cord blood CD34+ HSPC in vitro and study the SCID mice model of hematopoietic stem/progenitor cell (HSPC) transplantation. Methods The expression of objective gene in Ad-hLIF transgenic feeder cells was detected by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by magnetic-activated cell sorting(MACS) was detected by the flow cytometry. After expanded with various combinant of cytokines and transgenic feeder layer cells for 28 d, the quantity of mono-nuclear cell (MNC) and CD34+ cells rate was detected in different time. MNC after expansion stained by CFDA SE was injected to the sublethally irradiated SCID mice. Humanize gene Alu was detected by RT-PCR and fluorescence microscope. Results The green fluorescence was observed in the transgenic cells infected with 50MOI( multiplicity of infection) Ad-hLIF, and the objective gene was confirmed by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by MACS could reach 95.60% ±2.58%, Ad-hLIF transgenic feeder cells and various cytokines system increased MNC by 356.95±0.87 fold, and maximal expansion of CD34+ cells was observed during 0-14 d of culture, then down-expansion gradually. Four weeks after transplanted in SCID mice, fluorescently-labeled humanize cells still can be observed. The existence of the humanized gene Alu was confirmed by RT-PCR. Conclusion Ad-hLIF transgenic feeder cells can effectively proliferate umbilical cord blood CD34+ HSPC in vitro and delay it differentiate, what's more, it has high transplant efficacy and haematogenesis activity.

This paper is to establish a reversed-phase ion-pair chromatography (RP-IPC) method for universal estimation of the octanol/water partition coefficients (logP) of a wide range of structurally diverse compounds including acidic, basic, neutral and amphoteric species. The retention factors corresponding to 100% water (logk(w)) were derived from the linear part of the logk'/phi relationship, using at least four isocratic logk' values containing different organic compositions. The logk(w) parameters obtained were close to the corresponding logP values obtained with the standard "shake flask" methods. The mean deviation for test drugs is 0.31. RP-IPC with trifluoroacetic acid as non classic ion-pair agents can be applicable to determine the logP values for a variety of drug-like molecules with increased accuracy.
1-Octanol ; chemistry ; Chromatography, Reverse-Phase ; methods ; Hydrogen-Ion Concentration ; Octanols ; chemistry ; Trifluoroacetic Acid ; Water ; chemistry

Weight loss is frequently observed after acute exposure to high altitude. However, the magnitude and rate of weight loss during acute exposure to high altitude has not been clarified in a controlled prospective study. The present study was performed to evaluate weight loss at high altitude. A group of 120 male subjects [aged (32±6) years] who worked on the construction of the Golmud-Lhasa Railway at Kunlun Mountain (altitude of 4 678 m) served as volunteer subjects for this study. Eighty-five workers normally resided at sea level (sea level group) and 35 normally resided at an altitude of 2 200 m (moderate altitude group). Body weight, body mass index (BMI), and waist circumference were measured in all subjects after a 7-day stay at Golmud (altitude of 2 800 m, baseline measurements). Measurements were repeated after 33-day working on Kunlun Mountain. In order to examine the daily rate of weight loss at high altitude, body weight was measured in 20 subjects from the sea level group (sea level subset group) each morning before breakfast for 33 d at Kunlun Mountain. According to guidelines established by the Lake Louise acute mountain sickness (AMS) consensus report, each subject completed an AMS self-report questionnaire two days after arriving at Kunlun Mountain. After 33-day stay at an altitude of 4 678 m, the average weight loss for the sea level group was 10.4% (range 6.5% to 29%), while the average for the moderate altitude group was 2.2% (-2% to 9.1%). The degree of weight loss (Δ weight loss) after a 33-day stay at an altitude of 4 678 m was significantly correlated with baseline body weight in the sea level group (r=0.677, P<0.01), while the correlation was absent in the moderate altitude group (r=0.296, P>0.05). In the sea level subset group, a significant weight loss was observed within 20 d, but the weight remained stable thereafter. AMS-score at high altitude was significantly higher in the sea level group (4.69±2.48) than that in the moderate altitude group (2.97±1.38), and was significantly correlated with baseline body weight. These results indicate that (1) the person with higher body weight during stay at high altitude loses more weight, and this is more pronounced in sea level natives when compared with that in moderate altitude natives; (2) heavier individuals are more likely to develop AMS than leaner individuals during exposure to high-altitude hypoxia.
Adult ; Altitude ; Altitude Sickness ; physiopathology ; Body Mass Index ; Body Weight ; China ; Humans ; Hypoxia ; physiopathology ; Male ; Weight Loss ; physiology

In order to investigate the role of the hypoxia inducible factor 1 alpha (HIF-1α) in the adaptation mechanism to high altitude hypoxia, the cloning of the HIF-1α gene cDNA of Tibetan antelope (Pantholops hodgsonii), using RT-PCR and RACE, was applied, and the comparative analysis of the tissue-specific expressions of HIF-1α among Tibetan antelope, Tibetan sheep and plain sheep was performed using real-time PCR and Western blot. The sequence analysis indicated that the cDNA sequences acquired by cloning from the HIF-1α gene of Tibetan antelope comprised a 2 471-bp open reading frame (ORF) and a 1 911-bp 3'UTR. The similarity between its coding sequence, predicted amino acid sequence and HIF-1α of other mammals exceeded 87%, in which the similarity with cow was up to more than 98%, which showed that this sequence was the cDNA of HIF-1α of Tibetan antelope. The results of real-time PCR and Western blot showed that expressions of HIF-1α mRNA and protein appeared in Tibetan antelope's lung, cardiac muscle and skeletal muscle, with the highest expression in lung. HIF-1α mRNA and protein had obvious differential expression in these tissues. Further research showed that Tibetan antelope and Tibetan sheep possessed higher expressions of HIF-1α protein in the three tissues above-mentioned compared with plain sheep, and the expressions of HIF-1α mRNA and protein in Tibetan antelope's lung, cardiac muscle and skeletal muscle were higher than those of Tibetan sheep. It illustrates that the hypoxic HIF-1α-specific expression is one of the molecular bases of high altitude hypoxia adaptation in Tibetan antelope.
Adaptation, Physiological ; genetics ; physiology ; Altitude ; Animals ; Antelopes ; genetics ; physiology ; Cloning, Molecular ; Hypoxia ; physiopathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Lung ; metabolism ; Male ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Sheep ; Tibet

OBJECTIVETo evaluate the progression in morphologic changes of lungs in SARS patients.

METHODSFour cases of SARS with lung tissue samples available (including one for ultrastructural examination) were enrolled into the study. Histochemical study for VG, Masson, reticulin, orcein, PAS, sirius red stains and immunohistochemical study for vimentin, desmin, smooth muscle actin, HHF-35, CD34, F8, collagen types I and III were also performed.

RESULTSAccording to the morphologic changes, lung lesions in SARS were subcategorized into 3 phases: acute exudative inflammation, fibrous proliferation and the final fibrotic stage. Two cases belonged to the acute exudative phase, in which the course was less than 20 days. The principal lesions consisted of acute alveolar exudative inflammation, hyperplasia of alveolar epithelium, necrosis, alveolar hyaline membrane formation, alveolar desquamation and focal fibroplasia. The acute exudative protein was PAS-positive. There was an increase in reticulin fiber formation. The reactive fibroblasts were highlighted by desmin and vimentin. One case belonged to the fibroproliferative stage, in which the course was around 25 days. Major lesions included proliferative interstitial pneumonia with early pulmonary fibrosis. There was also evidence of organizing pneumonia, with an increase in reticulin fiber formation, which had a glomeruloid appearance on special stain. The mesenchymal cells showed either myofibroblastic (which expressed desmin, HHF-35, smooth muscle actin and vimentin) or fibroblastic (which expressed vimentin only) differentiation. Fibroelastosis and fibroplasia was also noted. The remaining case belonged to the fibrotic stage, in which the course was around 75 days. The main features included diffuse fibrosis and honeycomb change, which were highlighted by sirius red stain. Immunohistochemistry showed mainly types I and IV collagen fibers. In all lesions, there was also an increase of number of CD68-positive macrophages.

CONCLUSIONSThe morphologic progression in lungs of SARS patients is characterized by the development of increased fibrosis. The primitive mesenchymal cells, hyperplastic alveolar epithelial cells and macrophages play an important role in the pathogenesis.

Actins ; metabolism ; Adult ; Collagen Type I ; metabolism ; Desmin ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Male ; Middle Aged ; Pulmonary Fibrosis ; pathology ; Severe Acute Respiratory Syndrome ; metabolism ; pathology ; Vimentin ; metabolism

OBJECTIVETo study the status of iron deposition in patients with β-thalassemia intermedia and major in mainland China.

METHODSThe status of transfusion and chelation was examined in 39 patients with β-thalassemia intermedia or major. Serum ferritin levels were measured. MRI T2* technique was used to detect cardiac and hepatic iron deposition.

RESULTSSerum ferritin levels ranged from the minimum of 1500 ng/mL up to a maximum of 11491 ng/mL. From liver MRI T2* measurement, 15 cases had severe hepatic iron deposition (38%) and moderate deposition was found in 15 cases (38%), mild in 7 cases (18%), and normal in 2 cases (5%). Heart MRI T2* showed severe heart iron deposition in 7 cases (18%), mild in 5 cases (13%), and normal in 27 cases (69%). One case had cardiac arrhythmia. Four cases were over 20 years of age, and presented with gonadal function hypoplasia. The majority of patients did not receive regular transfusion and they had delayed, suboptimal chelation due to financial problems. Serum ferritin level was closely related with timing and dosage of chelation.

CONCLUSIONSIn patients with β-thalassemia who do not receive early regular transfusion and iron chelation therapy, iron deposition may occur at an early age. Important organs and tissue functional lesions and related complications also result. Relevant agencies and family members should be aware of this trend and develop appropriate strategies to improve the medical condition and quality of life of patients with this disorder.

Adolescent ; Adult ; Blood Transfusion ; Child ; Female ; Ferritins ; blood ; Humans ; Iron ; metabolism ; Liver ; metabolism ; Magnetic Resonance Imaging ; methods ; Male ; Myocardium ; metabolism ; beta-Thalassemia ; metabolism ; therapy