Aim The effect of tetrandrine on TGF-?1 mRNA expression in glomerulosclerosis rat was observed. Methods The rats were randomly divided into four groups, such as the normal control group (sham operative rat), glomerulosclerosis model group,tetrandrine group and amlodipine group. The expression of TGF-?1 mRNA was analyzed by Northern blot hybridization. Results The expressions of TGF-?1 mRNA in two treating groups were much lower than untreated model group. There were no difference between these two treating groups. Conclusion Tetrandrine can decrease the expression of TGF-?1 mRNA in glomerulosclerosis rat induced by unilateral renctomy plus adriamycin.

BACKGROUND:Hyperbaric oxygen (HBO) can increase oxygen diffusing capacity, thereby, improve hypoxic state of brain edema and brain tissue and promote the recovery of physiological function of brain cells in focal zone, the establishment of bypass circuit, and regeneration and repair of brain cells.OBJECTIVE: To observe the effect of hyperbaric oxygen on c-fos oncogene expression of rats at different time points following acute focal cerebral ischemia/reperfusion(I/R) injury.DESIGN : Randomized grouping animal experiment.SETTING: Department of Emergency, Tangdu Hospital, Fourth Military Medical University of Chinese PLA; Department of Laboratory Medicine, Xi'an Gaoxin Hospital;The General Hospital of the Air Force of Chinese PLA; Hyperbaric Oxygen Treatment Center, Department of Aerospace Medicine, Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was carried out in the Hyperbaric Oxygen Treatment Center, Department of Aerospace Medicine, Fourth Military Medical University of Chinese PLA in April 2002. Sixty-five 2-month-old healthy male SD rats.METHODS: The involved rats were randomized into: model group (n =20), normal control group (n =5), pure oxygen treatment group (n =20) and HBO treatment group (n =20). In the model group, following the method of Koizumi et al, rat models of middle cerebral artery (MCA) ischemia were developed. In the normal control group, only occlusion of arterial blood flow was omitted; In the pure oxygen treatment group, the operation procedure was the same as that of model group, and embolus being drawn out at ischemia for 1 hour, rats were placed in the hyperbaric cabin at 2,9,21, 45 and 69 hours after embolus being inserted, and they inhaled pure oxygen under the normal pressure; In the HBO treatment group, the operation procedure was the same as that of model group, and rats inhaled pure oxygen for 1 hour under 0.25 MPa pressure. MAIN OUTCOME MEASURES: By means of immunohistochemical and pathohistological methods, neutrophilic infiltration,c-fos oncogene protein and positive cell expression in cerebral cortex, preoptic area and corpora striatum of rats in each group were observed at cerebral I/R 5, 12, 24 and 72 hours; Neuronal necrosis degree in cerebral cortex, medial area of corpora striatum and preoptic area, and cerebrovascular leakage area of left cerebral hemisphere of rats were calculated.RESULTS: Sixty-five rats were involved in the final analysis. ① c-fos positive products mainly focused in the center of the preoptic area, but they were occasionally seen in the contralateral cortex, slightly expressed in the preoptic area and moderately expressed in the corpora striatum, c-fos positive products began to reduce in the above-mentioned area at ischemia 12 hours, and were obviously reduced at ischemia 24 hours; c-fos positive products in the cerebral cortex and preoptic area were obviously weakened in the HBO treatment group than in the simple ischemia group; At I/R 12 hours,neutrophils in the preoptic area and corpora striatum were significantly lower in the HBO treatment group than in the model group, respectively(P ＜ 0.05); At I/R 24 hours, neutrophils in the cerebral cortex, preoptic area and corpora striatum were significantly lower in the HBO treatment group than in the model group (P ＜ 0.05). ② Cerebrovascular leakage area was more significantly contracted in the HBO treatment group than in the model group (P＜ 0.05); At I/R 72 hours, the number of injured nerve cells in the optic chiasm cortex, medial area of corpora striatum and preoptic area was significantly smaller in the HBO treatment group than in the model group (P＜0.05). Neuronal damage was not found in the sham-operation group.CONCLUSION: HBO can markedly contract cerebrovascular leakage area of rats with acute focal cerebral ischemia/reperfusion injury, alleviate the symptoms of nervous system, inhibit neutrophilic infiltration and c-fos oncogene protein expression in the infarct area, and reduce neuronal necrosis in the "penumbral region".

BACKGROUND:Acute carbon monoxide (CO) poisoning may lead to delayed amnesia in rats,and which is similar to delayed neurologic syndrome caused by acute CO in human.So,this experiment is to investigate the pathogenesis of delayed neurologic syndrome by studying acute CO poisoning in the rats.OBJECTIVE:To observe the changes in delayed neuronal damage and memory after acute CO poisoning in the rats,and analyze their correlation.DESIGN:Randomized controlled animal experiment.SETTING:Department of Emergency,Tangdu Hospital,Fourth Military Medical University of Chinese PLA;Department of Laboratory Medicine,Xi'an Gaoxin Hospital;The General Hospital of the Air Force of Chinese PLA,Center for Hyperbaric Oxygen Treatment,Department of Aerospace Medicine,Fourth Military Medical University of Chinese PLA.MATERIALS:This experiment was carried out in the Laboratory of Aviation Pathology and Molecular Biology,Department of Aerospace Medicine.Fourth Military Medical University of Chinese PLA from July to November 2005.Fiftyhealthy male Sprague-Dawley(SD)rats were randomized into control group and CO poisoning group,with 25 rats each.METHODS:The awake rats in the CO poisoning group were placed in self-made jar for poisoning,then which was pumped with 0.999 volume fraction of CO.Rats in the jar inhaled the mixture of CO and air for 60 minutes.The average volume fraction of CO in the jar was 3.451×10-3.Rats in the control group were untouched.MAIN OUTCOME MEASURES:①The step down test was carried out in the rats before and 1,3,5 and 7 days after Coexposure.Escape latency was used as an index for evaluating the ability of memory retention.Shorter escape latencyindicated poor memory ability.②Pathological changes of brain tissue:After step down test was carried out following 1,3,5 and 7 days of CO exposure,6 rats were separately sacrificed in each group,and their brains were harvested.The brain tissue sections were performed haematoxylin & eosin (HE) staining for observing pathological injury degree and the amount of pyramidal neurons in hippocampal CA1 region.③SPSS 10.0 software was used to analyze the relationship of the amount of pyramidal neurons in hippocampal CA1 region and escape latency.RESULTS:Forty-eight rats were involved in the final analysis.①There were no significant differences in escape latencyon the 1"and 3"days after CO exposure between two groups. but escape latency in the CO poisoning group was significantly shorter than that in the control group on the 5th and 7th days after CO exposure(P＜0.05,0.01).②There were no significant changes in the amount of pyramidal neurons in hippocampal CA1 region on the 1st day after CO exposure between CO poisoning group and control group,but pyramidal neurons in hippocampal CA1 region in the CO poisoning group were significantly reduced on the 3rd,5th and 7th days after CO exposure,and 1 5%dead pyramidal neurons were found on the 7th day after CO exposure.③Decrease of pyramidal neurons in hippocampal CA1 region was significantly correlated with shortening of escape latency of rats in the CO poisoning group(r=0.270,P＜0.01).CONCLUSION:Acute CO poisoning leads to delayed neuronal damage,which causes delayed amnesia.

BACKGROUND: Nitric oxide (NO) plays an important role in the ischemic brain injury, and hyperbaric oxygen (HBO) can improve ischemia/reperfusion (I/R)-caused nerve injury. Whether the effect of HBO is associated with NO? Its mechanism needs to be further investigated.OBJECTIVE: To observe the changes of expression of nitric oxide synthase (NOS)-positive neurons of rats following acute focal cerebral I/R injury and HBO treatment.DESIGN: Randomized controlled animal experiment.SETTING: Department of Emergency, Tangdu Hospital, Fourth Military Medical University of Chinese PLA; Department of Laboratory Medicine, Xi'an Gaoxin Hospital; The General Hospital of the Air Force of Chinese PLA.MATERIALS : Sixty-six healthy male Sprague-Dawley rats were chosen and randomized into 5 groups: sham-operation group (n =5), sham-operation +HBO treatment group (n =5), model group (n =28), modeling +HBO treatment group (n =28). Ischemia 5,12, 24 and 72 hours four time points were set in the later 2 groups, 7 rats at each time point.METHODS: ①Rats in the model group and modeling+ HBO treatment group were created into models of middle cerebral artery ischemia according to the method from Koizum. Then, an embolus was inserted for ischemia; One hour later, the embolus was drawn out. Inserting embolus was omitted in the other two groups.②Rats in the sham operation + HBO treatment group and modeling + HBO treatment group were placed in HBO chamber at ischemia 2, 9, 21, 45 and 69 hours, separately, and given HBO treatment for 1 hour (0.25 MPa absolute pressure).MAIN OUTCOME MEASURES: The rats in each group were sacrificed at corresponding time points, and their brains were harvested. The distribution and morphology of NOS positive cells in cortical area, preoptic area, lateral and medial corpora striata of infarct region at the level of optic chiasma were observed with nicotinamide-adenine dinucleotide phosphate -diaphorase (NADPH-d) histochemical method.RESULTS: After supplement, 66 rats were involved in the final analysis. ①After ischemia, NOS-positive neurons changed in morphology, mainly presenting prominences were reduced or disappeared, neurons changed from ellipse or triangle into global shape, and shrank; Body of neuron darkly dyed; Both nucleus and cytoplasm were deeply dyed into dark blue; NOS-positive neurons with changed morphology were mostly in lateral corpora striatum, followed by preoptic area and medial corpora striatum, and those in the cortical area were few. NOS-positive neurons with changed morphology were not found in the sham-operation group and sham-operation + HBO treatment group. ②In the model group, NOS-positive neurons with changed morphology were increased with elongation of I/R time. At each time point, NOS-positive neurons in cortical area, preoptic area and medial corpora striatum in modeling + HBO treatment group were less than those in model group, but NOS-positive neurons in two groups both reached their peaks at ischemia 72 hours [Cortical area: (15.46±3.02) vs.(30.52±4.73)/visual field; Preoptic area:(28.56±4.05) vs. (68.81±7.84)/visual field; medial corpora striatum:(21.09±3.83) vs.(45.71±5.24)/visual field; all P＜0.01].CONCLUSION: HBO obviously inhibits the degeneration of NOS-positive neurons in acute focal cerebral I/R injury regions of rats, such as cortical area, preoptic area, medial corpora striatum, and so on

BACKGROUND:For pediatric patients with aplastic anemia in China, it is difficult to find human leucocyte antigen-matched sibling donors that are mostly replaced by parental donors.
OBJECTIVE:To retrospectively analyze the clinical efficacy and safety of parental haploidentical peripheral blood hematopoietic stem cel transplantation in children with relapsed and refractory severe aplastic anemia.
METHODS:Seventeen children with relapsed and refractory severe aplastic anemia who had no matched sibling or unrelated donor and failed to respond to immunosuppressive therapy were subjected to parental haploidentical peripheral blood hematopoietic stem cel transplantation. A conditioning regimen of fludarabine+cyclophosphamide+rabbit anti-human thymocyte immunoglobulin antibody and the triple therapy of methotrexate, cyclosporine A and mycophenolate mofetil were applied to prevent graft-versus-host disease.
RESULTS AND CONCLUSION: (1) Of the 17 children, 16 cases (94%) reached hematopoietic reconstitution, and the median time of neutrophils≥ 0.5×109/L and platelets≥ 20×109/L was 13 (11-15) days and 17 (12-28) days, respectively. (2) Incidence of acute graft-versus-host disease was 47% (8 of 17 cases), including 29% (5/17) of grades I-II and 18% (3/17) of grades III-IV. Incidence of chronic graft-versus-host disease was 41% (7/17). (3) With a median folow-up duration of 268 (43-753) days, the overal survival rate was 70.6% (12/17). Five dead cases (29%) belonged to transplantation-related death, including one case of fungal skin infections, one case of graft-versus-host disease, three cases of severe lung infection. No relapse case was reported. These findings indicate that if there are no matched sibling or unrelated donors and the immunosuppression effect is poor, parental haploidentical peripheral blood hematopoietic stem cel transplantation is a safe and effective salvage treatment for children with relapsed and refractory severe aplastic anemia.

OBJECTIVETo clone and sequence the cDNA of squalene epoxidase gene in Eleutherococcus senticosus.

METHODTotal RNA of E. senticosus was extracted by the improved isothiocyanate method and reverse transcripted into cDNA. The primers were designed depending on the reported SE cDNA sequences of Panax ginseng. The SE cDNAs in E. senticosus was amplified using RT-PCR strategy.

RESULTSequencing results showed two different cDNA fragments (SE1, SE2) with 1665, 1629 bp each ORF which encoded 554,542 amino acids, respectively. The identities of nucleotides and amino acids between SE1, SE2 were 91.49%, 92.55%. SE1, SE2 had the highest amino acids similarity to the SE1 of P. notoginseng, 93.45%, 94.87% respectively. SE1, SE2 both had a FAD binding domain. The deduced speculated amino acids of SE1, SE2 each had 2,4 membrane-spanning helices.

CONCLUSIONThe two SE sequences in E. senticosus were firstly separated and reported, which has made foundation for E. senticosus secondary metabolite engineering researches.

Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Eleutherococcus ; enzymology ; genetics ; Isoenzymes ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Squalene Monooxygenase ; classification ; genetics

To investigate the anti-tumor and side effect of CpG ODN 1826 and CpG ODN2006 as an adjuvants on leukemic tumor in mouse-models, an acute lymphocytic leukemic tumor in mouse model was established, then inoculated inactivated L1210 cells alone or with different vaccine adjuvants were injected subcutaneously into each DBA/2 model mouse at different times. The activities of mice, the tumor formation rate and the growth status of leukemic tumor were observed. The tumor was examined by pathologic section. The results showed that the vaccine of inactivated L1210 cells and CpG ODN 1826 could decrease the leukemic tumor formation rate, slow down the growth of leukemic tumor mass in mice and obviously cause necrosis of tumor cells, but it could not prolong the life spans of the tumor-burden mice; while CpG ODN2006 could not only decrease the tumor formation rate, slow down the growth of tumor mass in mice and result in obvious necrosis of tumor cells, but also could eliminate the existing tumor mass in mice, and prolong the life spans of the tumor-burden mice. It is concluded that using CpG ODN2006 as an adjuvant enhances the anti-tumor effect against the leukemic tumor, prolong the life span of tumor-burden mice without obvious side effect.
Adjuvants, Immunologic ; therapeutic use ; Animals ; Cancer Vaccines ; therapeutic use ; Cytotoxicity, Immunologic ; immunology ; Female ; Leukemia L1210 ; immunology ; therapy ; Male ; Mice ; Mice, Inbred DBA ; Neoplasm Transplantation ; Oligodeoxyribonucleotides ; therapeutic use ; Random Allocation

OBJECTIVETo compare the contents of baicalin in the raw medical material and the different prepared medical materials of Scutellaria baicalensis.

METHODThe contents of baicalin were determined by HPLC. Chromatographic conditions included Hypersil C18 column and the mobile phase consisting of a mixture of methanol-0.04% phosphoric acid (46:54). Baicalin was detected at 280 nm. The standard curve was linear in the range of 0.0280-4.6700 micrograms with correlation coefficient 1.0000. The average recovery of baicalin was 101.22% with RSD = 2.54% (n = 5).

RESULTThe contents of baicalin were 6.80% in raw material, 6.00% and 6.73% in the different prepared medical materials of S. baicalensis respectively.

CONCLUSIONThe two different kinds of preparing methods can be used to the raw material of S. baicalensis.

Chromatography, High Pressure Liquid ; Flavonoids ; analysis ; Hot Temperature ; Plants, Medicinal ; chemistry ; Scutellaria baicalensis ; chemistry ; Technology, Pharmaceutical

OBJECTIVETo compare the curative effects of primary osteoporosis treated with heat-sensitive point moxibustion and Gaitianli (Oyster Shell and Calcium Carbonate Chewable) tablets for oral administration and explore the treatment mechanism.

METHODSSixty cases of primary osteoporosis were randomly divided into a heat-sensitive point moxibustion group (moxibustion group) and a Gaitianli tablets group (medication group), 30 cases in each group. In the moxibustion group, the heat sensitized points were searched around Zusanli (ST 36), Pishu (BL 20), Shenshu (BL 23) and Mingmen (GV 4) and treated by heat-sensitive point moxibustion; in medication group, Gaitianli tablets were taken by oral administration, 3 pills for once and 3 times a day. The curative effects, bone mineral density (BMD), alkaline phosphatase (S-AKP) and urinary calcium to creatinine ratio (U-Ca/Cr) in both groups were observed before and after treatment.

RESULTSThe total effective rate was 86.7% (26/30) in moxibustion group, superior to that of 76.7% (23/30) in medication group (P < 0.05). After treatment, the BMD of lumbar vertebrae (L2-L4) mean was improved (P < 0.05), and the S-AKP and U-Ca/Cr were reduced (all P < 0.05); in medi cation group, the indexes above were no obvious changes (all P > 0.05).

CONCLUSIONThe therapeutic effect of primary osteoporosis treated with heat-sensitive point moxibustion is superior to that with Gaitianli tablets for oral administration. The mechanism is restraining bone resorption, increasing bone strength, keeping balance of bone metabolism, in order to increase bone mineral density and improve the clinical symptoms.

Acupuncture Points ; Aged ; Alkaline Phosphatase ; blood ; Bone Density ; Calcium ; urine ; Creatinine ; urine ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; methods ; Osteoporosis ; metabolism ; therapy

Objective To explore the influence of dexmedetomidine combined with sufentanil on postoperative analgesia effect and immune function for patients undergoing lower limb fracture surgery. Methods One hundred cases patients of lower limb fracture undergoing open reduction and internal fixation of elective surgery in our hospital from January 2016 to November 2016 were selected ( ASAⅠ-Ⅱgrade, male 51 cases and female 49 cases, 30-65 years old, 50-75 kg). All cases were randomly divided into sufentanil group (Group S) and dexmedetomidine combined with sufentanil group (Group DS) according to the random number table, who used patient-controlled intravenous analgesia (PCIA). The PCIA drug formulations of two groups were as followings: sufentanil group (Group S) used 3 μg/h sufentanil+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;dexmedetomidine combined with sufentanil group (Group DS) used dexmedetomidine 0.1μg/(kg · h)+sufentanil 2μg/h+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;background infusion rate was 2 ml/h, and loading dose was 2 ml. Dose for patient-controlled analgesia (PCA) was 0.5 ml, and locking time was 15 min. The changes of pain, MAP, HR , sedation scores were recorded at 4 h (T1), 8 h (T2), 24 h (T3) and 48 h (T4) after operation; nausea and vomiting, hypotension, bradycardia, respiratory depression and other adverse reactions were aslo recorded at the same time. At 10 min before induction of anesthesia (T0) and T1-T4 after operation, the CD3+, CD4+, CD8+and NK cell activity of peripheral blood T lymphocyte subsets were determined using flow cytometry, and CD4 +/CD8 + ratio was calculated. Results The levels of MAP and HR in DS group at each time point after operation were lower than those in group S (P<0.05), the level of visual analogue score (VAS) in DS group at different time points were significantly lower than those of s group: (1.8 ± 0.3) scores vs. (2.5 ± 0.5) scores, (1.1 ± 0.5) scores vs. (1.9 ± 0.3) scores, (1.0 ± 0.5) scores vs. (1.8 ± 0.5) scores, (0.8 ± 0.3) scores vs. (1.5 ± 0.6) scores (P < 0.05), Ramsay Sedation score was significantly higher than that of s group: (3.5 ± 0.3) scores vs. (2.4 ± 0.6) scores, (3.3 ± 0.5) scores vs. (2.5 ± 0.3) scores, (3.5 ± 0.6) scores vs. (2.3 ± 0.5) scores, (3.2 ± 0.4) scores vs. (2.2 ± 0.5)scores (P<0.05);nausea and vomiting occurred in two patients after the operation of group DS , which was significantly lower than that of S group (P<0.05);compared with those at T0, CD3+, CD4+, CD4+/CD8+and NK cells in the two groups of patients decreased significantly at the time of T1-T4 (P<0.05);The levels of CD3+, CD4+, CD4+/CD8+and NK cells were significantly higher in group DS at T1- T4 than those in group S(P < 0.05). Conclusions Dexmedetomidine combined with sufentanil for analgesia in patients with lower limb fracture surgery has better effect. It decreases the incidence of nausea and vomiting, and improves the cellular immune function of patients.