OBJECTIVETo explore the molecular biological mechanism of acupuncture and moxibustion (A&M) in reducing chemotherapy-related toxicity, relieving myelosuppression and increasing peripheral white blood cells (WBC).

METHODSTwo hundred and twenty-four male Kunming mice of clean grade were randomized equally into 4 groups, the blank control group (A), the model group (B), the acupuncture group (C), and the moxibustion group (D). Except those in Group A, mice were duplicated into myelosuppression model with cyclophosphamide (CTX) using the accepted method. After being modeled, mice in Group C and D were treated with acupuncture and moxibustion respectively, once a day for 7 successive days, while those in Group A and B were dealt with the same actions (seizing and fixing) every day but no therapy was given. From day 2 to day 7 of the treatment, 8 mice were taken from every group per day and killed in batches. Their peripheral WBC was counted and bone marrow for detecting Cyclin D1 expression and percentages of bone marrow cells in different cycle stages using immunohistochemistry and flow cytometer respectively.

RESULTSWBC count restored to exceed the baseline in group C and D at day 5 of the treatment, being one day earlier than that in group B. Cyclin D1 expression in the bone marrow raised in Group C and D, and reached the peak at day 4, showing significant difference as compared with that in Group B (P < 0.01). The phase G1 marrow cell percentages in Group C and D was lower than that in Group B at all days of detection, showing statistical significance at day 2-4 (P < 0.05 or P < 0.01); while the percentages of phase S and G2-M cells in the two treated groups was higher than that in group B all the times.

CONCLUSIONSWhile CTX damaged marrow cells, it intervened the cell cycle regularity and reduced the DNA content to cause myelosuppression and leucocytopenia. A&M therapy could improve the Cyclin D1 expression, speed up the cell transition from phase G1 to phase S and increase the synthesis of DNA. At the mean time, taking advantage of the block at phase G2, it can repair the injured DNA, speed up cell mitosis for turning into multiplication, improve the anti-injury and repairing ability of cells to protect bone marrow cells, and relieve the myelosuppression.

Acupuncture Therapy ; Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cyclin D1 ; metabolism ; Cyclophosphamide ; adverse effects ; Immunosuppressive Agents ; adverse effects ; Male ; Mice ; Mice, Inbred Strains ; Moxibustion

Based on the full-length cDNA of BmalphaTX14 from Chinese scorpion Buthus martensii Karsch (BmK), gene of the mature peptide of BmalphaTX14 was cloned into the yeast expression vector pPIC9K. After transforming, screening and inducing, tricine-SDS-PAGE and Western blot proved that rBmalphaTX14 protein was expressed in the medium for up to 84 hours, getting nearly 120 mg/L. Recombinant BmalphaTX14 was purified rapidly and efficiently through Ni-NTA-agarose, polyethylene glycol precipitation and gel filtration chromatography. The purified rBmalphaTX14 proved to have the anti-insect activity by toxicity assay. Meanwhile, genomic gene of BmalphaTX14 was cloned and sequenced by PCR method, sequence analysis of this gene showed that BmalphaTX14 had an intron of 408 base pairs located at the signal peptide encoding region, which was similar with the characteristic of other alpha-type sodium ion-channel toxin. Considering both the genomic organization and the peptide function, BmaTX14 proved to be a membership belonging to alpha-type sodium ion-channel toxin.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Scorpion Venoms ; biosynthesis ; genetics ; Scorpions ; genetics ; Sequence Analysis

OBJECTIVETo enlarge the ratio of length to width of a prefabricated random skin flap in a short period, in order to meet a special clinical need.

METHODSA white rat and a white mini-pig was chosen for an animal model for the experiment. They were divided into the experimental group and the control. The prefabricated flap was formed as the planned design. In the experimental group, the flaps were treated with a desired pressure in certain intervals at the planned part of the flap. We estimated and inspected the digitized perfusion of flap microcirculation, and made overcong and wide random skin flap survive within 24 hours, and this technique was finally used for the clinical wound treatment.

RESULTSAs expected, the survival area of the flap in the experimental group was significantly larger than the control. This technique was successfully used to repair the defects of the special sites in plastic and reconstructive surgery.

CONCLUSIONThe method of quickly prefabricated random flap could enlarge the ratio of length to width of the flap and put the flap in shape in short time. With the digitized estimating and inspecting, this method could obviously improve the survival area of the random flap and could also be very useful for the clinical treatment.

Adult ; Animals ; Dermatologic Surgical Procedures ; Female ; Humans ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; pathology ; Skin Transplantation ; methods ; Surgical Flaps ; supply & distribution ; Time Factors ; Transplantation, Heterologous ; Treatment Outcome

To investigate the eletrophysiological effect of rat adrenomedullin (rADM) on barosensitive neurons in the rostral ventrolateral medulla (rVLM) and its potential mechanisms, the extracellular recording and multi-barrel iontophoresis methods were used. Of the 29 barosensitive neurons in the rVLM, 20 neurons demonstrated excitatory response to iontophoretically applied rADM and increased the firing rate from (10.8 +/- 2.7) spikes/s to (14.6 +/- 3.6), (19.8 +/- 4.7) and (31.9 +/- 6.4) spikes/s (P<0.05, n=20) at the current of 30, 60 and 90 nA, respectively. Application of human adrenomedullin (22-52) [hADM (22-52)], a specific antagonist of rADM receptor, distinctly attenuated the augmentation of firing rate induced by rADMjthe firing rate was increased by 15.4% [(11.4 +/- 2.5) spikes/s, P<0.05, n=10]. Another antagonist, human calcitonin gene-related peptide (8-37) [hCGRP (8-37)] had no significant effect on rADM-induced excitation. Other 23 barosensitive neurons were recorded to test the influence of nitric oxide synthase (NOS) inhibitors on the excitatory effect of rADM. In 10 neurons, 7-NiNa (neuronal NOS inhibitor) decreased the firing rate from (10.1 +/- 3.5) spikes/s to (7.5 +/- 2.5), (5.3 +/- 2.1) and (3.1 +/- 1.4) spikes/s (P<0.05, n=10) at the current of 10, 20 and 40 nA, respectively. The excitatory effect of rADM (60 nA, 30 s) during 7-NiNa application was nearly eliminated and the magnitude of firing rate was increased only by 17% of the basal level (6.2 +/- 1.9) spikes/s (P<0.05, n=7). While aminoguanidine (AG, iNOS inhibitor) increased the firing rate at the resting level from (11.5 +/- 5.1) spikes/s to (17.8 +/- 5.6), (22.5 +/- 6.3) and (29.1 +/- 6.4) spikes/s (P<0.05, n=8) at the current of 10, 20 and 40 nA in 8 barosensitive neurons, respectively. When rADM (60 nA, 30 s) was delivered during AG iontophoresis period, the firing rate significantly increased by 60% of the basal level [(22.5 +/- 6.3) spikes/s, n=5]. These results indicate that rADM activates the barosensitive neurons in the rVLM directly and acts as a cardiovascular regulator, and that this function might be mediated by its specific receptor. NO, mainly neuronal NOS-originated might be involved in the excitatory effect of rADM in the rVLM.
Adrenomedullin ; physiology ; Animals ; Electrophysiological Phenomena ; Male ; Medulla Oblongata ; physiology ; Neurons ; physiology ; Nitric Oxide ; physiology ; Nitric Oxide Synthase Type I ; physiology ; Pressoreceptors ; physiology ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the effects and the impact on maternal and infant outcome of Tianma-Gouteng Decoction combined low-dose aspirin for high-risk patients with pre-eclampsia (PE). Methods:A total of 94 high-risk patients who met the diagnose cretieria of PE from January 2017 to January 2019 in our hospital were divided into two groups by the random number table method, each group 47 cases. The control group based on symptomatic treatment, received daily 75 mg of aspirin enteric-coated metformin hydrochloride additionally; and the observation group took Tianma-Gouteng Decoction based on the control group. Both groups stopped taking medicine after 28th week of gestation. Immune turbidimetric method was used to detect D-dimer (D-D), the Claus method was used for detecting fibrinogen (Fib). The blood pressure meter was used to measure systolic blood pressure (SBP) and diastolic blood pressure (DBP). The poor maternal and infant outcomes were recorded. Results:After the treatment, the plasma D-D (0.50 ± 0.12 μg/L vs. 0.60 ± 0.30 μg/L; t=2.122, P=0.037), Fib (2.37 ± 0.22 g/L vs. 4.43 ± 0.56 g/L; t=23.473, P<0.01) in the observation group were significantly lower than the control group. The SBP (83.43 ± 4.01 mmHg vs. 96.30 ± 5.34 mmHg, t=13.212), DBP (113.77 ± 3.90 mmHg vs. 126.52 ± 4.43 mmHg, t=14.810) in the observation group were significantly lower than the control group ( P<0.01). The incidence of fetal and neonatal adverse outcomes in the observation group was 10.6% (5/47), the control group was 25.5% (12/47), and there was statistically significant between two groups ( χ2=3.519, P<0.05). The incidence of adverse outcome in the observation group was 31.9% (15/47), the control group was 55.3% (26/47), and there was statistically significant between two groups ( χ2=4.326, P<0.01). Conclusions:Tianma-Gouteng Decoction combined low-dose aspirin enteric-coated metformin hydrochloride can improve coagulation function, lower blood pressure, improve clinical efficacy, reduce the occurrence of advers outcomes for high-risk patients with PE.

Objective To explore the factors associated with the complications of cranioplasty, infection and local epidural hematoma, such as age, sex, primary disease, skull defect and concomitant hydrocephalus. Methods 211 patients after cranioplasty were reviewed. Results 8 cases (3.8%) complicated one of them, in which 6 cases with local epidural hematoma and 2 cases with infection. The infection was more likely related with the areas of skull defect (P=0.003). No factor was found related with the local epidural hematoma. Conclusion It is necessary to focus the size of the frontal sinus when repairing the the frontal bone, to avoid the screw into the frontal sinus caused infection.

Objective To study the content of serum granulocyte macrophage colony stimulating factor(GM-CSF)and granulocyte colony stimulating factor(G-CSF)in cyclophosphamide(CTX)-induced tumor-bearing mice before and after acupuncture and moxibustion treatment,so as to find out the mechanism of acupuncture and moxibustion treatment on recovery from myelosuppression and increasing of leukocyte counts.Methods 64 Kunming male mice of clean grade were transplanted subcutaneously S180 sarcoma cells to induce the trans-planting tumor model,then were randomly divided into control group,model group,acupuncture group and moxibustion group (each n =16)according to the body weight.Mice in model group,acupuncture group and moxibustion group were intraperitoneally injected with CTX to establish CTX-induced tumor-bearing mice mod-el,and the control group were injected intraperitoneally with normal saline.Acupoints Dazhui(GV14),Geshu (BL17),Shenshu (BL23)and Zusanli (ST36)were needled in acupuncture group and were moxa moxibusted in moxibustion group once daily at regular time for consecutive 5 d.Peripheral leukocyte counts were meas-ured by detecting the blood collected from caudal vein before modeling and at every day of treatment proce-dure.The serum concentrations of GM-CSF and G-CSF were determined by using ELISA method at the third day of treatment and one day after five-day-treatment.Results Compared with control group,peripheral leu-kocyte counts decreased,even to the lowest level at the fourth day in the three groups of CTX-induced tumor-bearing mice,and the serum concentrations of GM-CSF and G-CSF also decreased in the three groups signifi-cantly(P <0.05).At the fifth day of treatment,leukocyte counts started to increase,and the serum concen-trations of GM-CSF and G-CSF increased significantly compared with model group(P <0.05).At the day after treatment,leukocyte counts increased to nearly normal level and the serum concentrations of GM-CSF and G-CSF increased dramatically compared with model group.Conclusion CTX damaged the myeloid element to reduce peripheral leukocyte counts and the serum concentrations of GM-CSF and G-CSF.Acupuncture and moxibustion can obviously increase the concentration of GM-CSF and G-CSF,that is to say,both the Chinese medical treatment can induce colony growth of the granulocyte precursor and macrophage precursor cells and stimulate differentiation of the hematopoietic cells,leading to proliferation,differentiation and maturation of the myeloid hematopoietic cells,resulting in recovery from myelosuppression and increasing of leukocyte counts induced by chemotherapy.

Abstract:One H5N1 subtype avian influenza virus, A/duck/Shandong/009/2008 (Dk/SD/009/08), was isolated from apparently healthy domestic ducks in some live bird market in East China during our epidemiological surveillance. To investigate the genetic composition, Dk/SD/009/08 was subjected to genome sequencing. The amino acid motif of cleavage site was "PLRERRRK-R/GL", which was consistent with the characterization of the HPAIV. According to the newest unified nomenclature system of H5N1, Dk/SD/ 009/08 was classified into Clade 2.3.4. The BLAST results showed that four gene segments (HA, NA, NP and NS) had the highest nucleotide identities with H5N1 subtype AIVs whereas the remaining four (PB2, PB1, PA and M) displayed the closest relationship with H9N2 subtype. Therefore, Dk/SD/009/08 might be a natural reassortant virus. The phylogenetic analysis further indicated that G1-like H9N2 subtype AIVs which was prevalent mainly in quails of Southern China might provide the internal genes for Dk/ SD/009/08.
Animals ; Chick Embryo ; Genome, Viral ; Humans ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza A Virus, H9N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Molecular Sequence Data ; Phylogeny ; Reassortant Viruses ; classification ; genetics ; isolation & purification ; Recombination, Genetic ; Viral Proteins ; genetics

Methylation, a methyl group-consuming reaction, plays a key role in the degradation (i.e., inactivation) of monoamine neurotransmitters, including catecholamines, serotonin and histamine. Without labile methyl groups, the methylation-mediated degradation cannot take place. Although high niacin (nicotinic acid and nicotinamide) intake, which is very common nowadays, is known to deplete the body's methyl-group pool, its effect on monoamine-neurotransmitter degradation is not well understood. The aim of this article was to investigate the effect of excess nicotinamide on the levels of plasma serotonin and histamine in healthy subjects. Urine and venous blood samples were collected from nine healthy male volunteers before and after oral loading with 100 mg nicotinamide. Plasma N(1)-methylnicotinamide, urinary N(1)-methyl-2-pyridone-5-carboxamide (2-Py), and plasma betaine levels were measured by using high-performance liquid chromatography (HPLC). Plasma concentrations of choline, serotonin and histamine were measured using commercial kits. The results showed that the plasma N(1)-methylnicotinamide level and the urinary excretion of 2-Py significantly increased after oral loading with 100 mg nicotinamide, which was accompanied with a decrease in the methyl-group donor betaine. Compared with those before nicotinamide load, five-hour postload plasma serotonin and histamine levels significantly increased. These results suggest that excess nicotinamide can disturb monoamine-neurotransmitter metabolism. These findings may be of significance in understanding the etiology of monoamine-related mental diseases, such as schizophrenia and autism (a neurodevelopmental disorder).
Betaine ; blood ; Choline ; blood ; Chromatography, High Pressure Liquid ; Histamine ; blood ; Humans ; Male ; Niacinamide ; administration & dosage ; analogs & derivatives ; blood ; Pyridones ; urine ; Serotonin ; blood

OBJECTIVETo explore the molecular biological mechanism of acupuncture and moxibustion for relieving myelosuppression and increasing white blood cells.

METHODSTwo hundred and twenty-four clean male Kunming mice were randomly divided into a control group, a model group, an acupuncture group and a moxibustion group, 56 mice in each group. The model of myelosuppression was made with Cyclophosphamide. In the acupuncture group and the moxibustion group, acupoints "Dazhui" (GV 14), "Geshu" (BL 17), "Shenshu" (BL 23) and "Zusanli" (ST 36) were used for treatment with acupuncture and moxibustion, respectively, while, in the control group and the model group, there were no treatment carried out except catching and fixing. The changes of bone marrow cell DNA pol beta and XPD between the 2nd and 7th day were examined with immunohistochemical method.

RESULTSAcupuncture and moxibustion markedly up-regulated the expression of bone marrow cell DNA pol beta and XPD, and promoted the base excision repair and nucleotide excision repair, which leads to the relieving Cyclophosphamide-induced myelosuppression and increasing the number of white blood cells.

CONCLUSIONFor acupuncture and moxibustion, one of the bone major mechanisms in relieving post-chemotherapy myelosuppression, protecting hemopoietic function and increasing the white blood cells is that it can promote the repair of the bone marrow cell DNA excision and protect hemopoietic cells from injury by chemical drugs.

Acupuncture Therapy ; Animals ; Bone Marrow Cells ; enzymology ; Cyclophosphamide ; pharmacology ; DNA Damage ; drug effects ; DNA Repair ; DNA Repair Enzymes ; genetics ; metabolism ; Gene Expression ; drug effects ; Male ; Mice ; Moxibustion ; Random Allocation