Preliminary research in our laboratory found that compound YZG-330 can reduce mouse body temperature, which could be blocked by adenosine A₁ receptor (A₁R) antagonist DPCPX. Based on the downstream signaling pathway of the A₁R, the mechanism by which YZG-330 lowers body temperature was further studied. The pharmacodynamics of YZG-330 was evaluated by measuring the rectal temperature; expression of the transient receptor potential (TRP) ion channel, the P38 protein and its phosphorylated form in mouse hypothalamic homogenate were detected by Western blotting. A Ca²⁺ fluorescent probe, Fluo-3AM, was added to cells to detect the effect of YZG-330 on the Ca²⁺ content of mouse hypothalamic cells. YZG-330 dose-dependently reduced the body temperature in mice, and the selective P38 inhibitor SB-203580 (20 mg·kg^-1, i.p.) significantly inhibited the hypothermic effect of YZG-330. A TRPM8 antagonist 2 (0.1 μg per mouse, i.c.v.) markedly attenuated the hypothermic effect of YZG-330 (0.25 or 1 mg·kg^-1, i.p.). YZG-330 (2 mg·kg^-1, i.p.) significantly increased the phosphorylation of P38, an effect that could be attenuated by the A₁R antagonist DPCPX (5 mg·kg^-1, i.g.) in mouse hypothalamus. In addition, YZG-330 also prominently enhanced the expression of TRPM8, which could be blocked by SB-203580; YZG-330 (0.1-10 μmol·L^-1) increased intracellular Ca²⁺ concetration in mouse hypothalamic cells in a dose-dependent manner, and was inhibited by the A₁R inhibitor DPCPX (0.5 and 1 μmol·L^-1) and TRPM8 antagonist 2 (1 μmol·L^-1). In conclusion, YZG-330 exerts its hypothermic effect by activating the A₁R to promote the phosphorylation of P38 protein and thereby up-regulating the expression and activity of the TRPM8 ion channel, resulting in increased intracellular Ca²⁺ concentration to stimulate mouse hypothalamus cells to down-regulate body temperature. All animal experiments were approved by the Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences.

OBJECTIVETo explore the clinical effect of the sacrococcygeal space injection for the treatment of failed back surgery syndrome.

METHODSFrom July 1998 to October 2012,47 patients with failed back surgery syndrome were treated and included 39 males and 8 females with an average age of 61.5 years old ranging from 35 to 89 years old. Among them,41 patients experienced one time of operation, 6 patients with twice of operation. Forty-one patients underwent single,bilateral fenestration or central laminectomy decompression, discectomy. Six patients underwent total laminectomy discectomy and inter body fusion and pedicle screw fixation. All patients were examined by X-ray plain film, CT or MRI before treatment. The anticoagulation was discontinuation before treatment. The needle was put into the sacrococcygeal gap at prone position in the sense of frustration,suction without cerebrospinal fluid and blood,with injection of Mailuoning (Chinese characters: see text) 15 ml. The pain was assessed by VAS before and after treatment. The Oswestry low back pain disability index and survival quality interference degree were evaluated.

RESULTSAt 1 month after treatment,the pain VAS decreased from 59.24 +/- 17.35 before treatment to 19.19 +/- 11.19 after treatment (P < 0.05); The Oswestry low back pain disability index decreased from (41.35 +/- 9.87)% before treatment to (23.17 +/- 17.56)% after treatment (P < 0.05); The survival quality interference degree decreased from 6.5 +/- 2.2 before treatment to 2.6 +/- 1.4 after treatment (P < 0.05).

CONCLUSIONThe sacrococcygeal gap injection for treatment of failed back surgery syndrome has advantages of simple, safe, fewer complications, and low treatment cost.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Failed Back Surgery Syndrome ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Sacrococcygeal Region ; diagnostic imaging

Adult ; Granuloma, Respiratory Tract ; microbiology ; Humans ; Male ; Mycoses ; diagnosis ; therapy ; Pharyngeal Diseases ; diagnosis ; microbiology ; therapy

OBJECTIVE: Mitochondrial deoxyribonucleic acid (mtDNA) 8344 A>G (m.8344A>G) mutation is the common mutation associated with mitochondrial myoclonus epilepsy with ragged-red fibers (MERRF) syndrome. Herein we report a rare case with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes/MERRF/Leigh (MELAS/MERRF/Leigh) overlap syndrome caused by m.8344A>G mutation. METHODS: The clinical and imaging data of the patient were collected and an open muscle biopsy was carried out. We further employed molecular genetic analyses to detect mtDNA mutation in the proband and his mother. And then a clinical and neuroimaging follow-up was performed. RESULTS: This patient was a 25-year-old male, who developed exercise intolerance since the age of 6. At age 10, he suffered from acute episodes of hemianopia, and cranial magnetic resonance imaging (MRI) showed occipital stroke-like lesions and cranial magnetic resonance spectroscopy (MRS) revealed a lactate peak corresponding to the lesion. After that the patient presented slowly progressive psychomotor decline. He had myoclonic seizures and cerebellar ataxia since the age of 12. At age 21, he was admitted to our hospital because of confusion and cranial MRI revealed symmetrical lesions in bilateral posterior putamen, thalami and midbrain. Then repeated MRI showed progression of original lesions and new frontal multiple stroke-like lesions. Symptomatic and rehabilitation treatment relieved his condition. Follow-up cranial MRI at age 24 showed the lesions in basal ganglia and thalami diminished, and the midbrain lesions even completely vanished. Muscle pathology indicated the presence of numerous scattered ragged-red fibers (RRF), suggestive of a mitochondrial disorder. Polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) detected the m.8344A>G mutation of the MT-TK gene encoding mitochondrial transfer RNA for lysine in the patient's blood. Next generation sequencing (NGS) of the whole mitochondrial genome identified that the proportion of m.8344A>G was 90%, and no other mtDNA mutation was detected. Sanger sequencing further identified this mutation both in the proband and his mother's blood, although the mutation load was much lower in his mother's blood with approximately 10% heteroplasmy. CONCLUSION The present study is the first to describe a patient with m.8344A>G mutation in association with the MELAS/MERRF/Leigh overlap syndrome, which expands the phenotypic spectrum of the m.8344A>G mutation.
Acidosis, Lactic ; Adult ; Child ; DNA, Mitochondrial/genetics* ; Humans ; Male ; Mitochondrial Encephalomyopathies ; Mutation ; Stroke ; Young Adult

The traditional Chinese medicine of Radix Hedysari plays an important role in invigorating gas for as-cending, benefiting blood for promoting production of fluid, and promoting circulation for removing obstruction in collaterals, which is consistent with the principle of treatment for osteoporosis. This study is designed to investigate the bioactive components on increasing peak bone mass (PBM) by exploring the spectrum-effect relationship between chromatography fingerprints and effect. Multiple indicators are selected to evaluate the pharmacological activity. In fingerprints, 21 common peaks are obtained, five of which are identified. Furthermore, gray relational analysis (GRA) is a quantitative method of gray system theory and is used to describe the correlation degree of common peaks and pharmacological activities with relational value. 21 components are then divided into three different regions, of which ononin and calycosin play an extremely significant role in increasing PBM. In addition, factor analysis and hierarchical cluster analysis (HCA) are used to screen the optimal producing area for Radix Hedysari. This provides a comprehensive and efficient method to improve the quality evaluation of Radix Hedysari, confirming the bioactive components for PBM-enhancement and further develop its medicinal value.

OBJECTIVETo establish a retinal vein occlusion (RVO) animal model and observe the therapeutic effect of a Chinese herbal composition (Fufang Xueshuantong Capsule, , FXC)inischemicinischemic) in ischemic retinal disease.

METHODSFifteen adult male Sprague-Dawley rats underwent laser photothrombosis to induce RVO on their right eyes and were subsequently randomized to receive FXC (the intervention group, n=7) or placebo treatment (the control group, n=8). Fundus fluorescein angiography was performed after 2, 4 and 8 weeks of treatment. Real-time reverse transcription-PCR was used to quantify the mRNA expression of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1). The main outcomes were the mRNA copies of VEGF and SDF-1 and the counts of RVO signs.

RESULTSLaser photothrombosis procedure induced typical lesions of RVO, including hemorrhage, leakage, retinal detachment, capillary non-perfusion, filling defect of retinal vessels, and lateral circulation/dilation of small vessels. The retinal lesions were associated with an increased expression of VEGF (P<0.05). No significant change of SDF-1 expression was noticed. Compared with the control group, the intervention group had numerically fewer RVO lesions at week 2 (1.71±0.76 vs. 3.50±1.51, t=-2.82, P<0.05). The benefit of intervention remained at weeks 4 and 8.

CONCLUSIONSA rat model of laser photothrombosis-induced RVO was established and an increase in the VEGF expression was observed in the retinal lesion. The FXC had therapeutic benefit in improving retinal lesions in the rat model of RVO.

Animals ; Base Sequence ; Chemokine CXCL12 ; genetics ; metabolism ; DNA Primers ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Fluorescein Angiography ; Gene Expression ; Male ; Rats ; Rats, Sprague-Dawley ; Retinal Vein Occlusion ; metabolism ; therapy ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

OBJECTIVETo find a new and effective way for the transfection of adenovirus vectors encoding HSP70 cDNA, so as to provide another possible method in gene therapy against ischemia and cellular hypoxia after burn injury.

METHODSThe replicated defective adenovirus vectors encoding HSP70 cDNA were encapsulated. Its acid resistance and dissolution in intestinal fluid were tested in artificial gastric juice and intestinal fluid. The expression of HSP70 gene which was transfected by the microcapsules orally was detected by RT-PCR.

RESULTSThe encapsulated replicated defective adenovirus vectors were viable in vitro. They exhibited good resistant to acid (resolution ratio less than 10%) and dissolution in intestinal juice (resolution ratio higher than 50%). The HSP70 gene expression of the tested rats was significantly higher than control, but there was no difference in the quantity of HSP70 induced by sodium arsenite or adenovirus transfection through injection by vein.

CONCLUSIONThe encapsulation of adenovirus vectors can successfully keep the viability of the virus in vitro and protect the virus from harmful effect of acid and enzyme in the gastric juice. Its nice dissolution in intestinal juice should ensure its absorption by oral transfection. The expression of the HSP70 gene after oral intake of this preparation is as high as that with other traditional transfection methods. It is possible that in the future the encapsulated replication of defective adenovirus vectors encoding HSP70 cDNA can provide a safer, convenient and effective way for gene therapy for burn patients.

Adenoviridae ; genetics ; Animals ; Burns ; complications ; Capsules ; Genetic Therapy ; Genetic Vectors ; HSP70 Heat-Shock Proteins ; administration & dosage ; genetics ; Hypoxia ; etiology ; prevention & control ; Intestinal Mucosa ; metabolism ; Ischemia ; etiology ; prevention & control ; Male ; Myocardium ; metabolism ; Rats ; Rats, Wistar ; Transfection

Objective　To study the effect of acute serious smoke inhalation injury to unilateral lung on contralateral lung and the therapeutic efficacy of massive bronchoscopic lavage with saline to the primary injured bronchoalveolus at early stage after smoke inhalation on mitigating the secondary damage of the other lung and so as to confirm the therapeutic validity of lung lavage to smoke inhalation injury. Methods　Fifteen mongrel dogs were insufflated with sawdust smoke into left lung and then randomized into 2 groups. The dogs in group A were maintained intravenous glucose saline transfusion and breathed air freely for 24 h after injury. Those in group B received a massive bronchoscopic lavage with 250 ml of saline to injured lung at 1 hour after injury and then were administrated similarly to those in group A. Gas exchange function of bilateral lungs in the process were observed and pathologic and phyiopathologic changes of the lung specimen were examined after the process. Results　While the total lavaging volume was kept to 250 ml, the liquid volume remained in left lung after each operating was from 90 ml to 140 ml. All animals had a significant decline in pulmonary function after smoke inhalation injury. PaO2 in group B decreased significantly soon after the lavage and then increased gradually to exceeding that in group A. The left lung showed serious edema similarly in two groups but in right lung the water content was less, dynamic and static compliance was more in group B than in group A. Conclusion　Bronchoalveolar massive lavage at early post-injury stage to injured lung after unilateral lung smoke inhalation injury can mitigate secondary damage of the non-injured lung. Lung lavage can decrease the intensity of secondary systemic inflammatory reaction and show a therapeutic validity to smoke inhalation injury.

OBJECTIVETo use heme oxygenase (HO) inducer hemin and a HO inhibitor zinc protoporphyrin (ZnPP) to investigate the effect of HO on apoptosis and apoptosis genes in hepatic ischemia reperfusion (IR) injury in rats.

METHODSNinety-six Sprague-Dawley rats were randomly divided into four groups (24 rats in each): a sham-operation group, an ischemia-reperfusion (IR) group, a hemin-IR group and a ZnPP-IR group. Liver functions, liver histology and hepatocellular apoptosis rates were observed at 0, 1.5, 4 and 8 hours after reperfusion. Hepatocellular apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; expressions of Bcl-2 and caspase-3 were determined by Western blot.

RESULTSCompared with the sham-operation group, the levels of ALT and AST were increased in the IR group. In the IR group the histological changes found in the livers were swelling of hepatocytes, narrowing of hepatic sinusoids, inflammatory cell infiltration and necrosis of hepatocytes in some areas of the livers. In the IR group rate of hepatocellular apoptosis was increased at 0, 1.5, 4 and 8 hours after reperfusion; expression of Bcl-2 was decreased and the expression of caspase-3 was increased. In the hemin-IR group, the levels of ALT and AST were lower, the pathological changes were milder and the rate of hepatocellular apoptosis was lower at 0, 1.5, 4 and 8 hours in comparison to those of the IR group. The expression of Bcl-2 was higher and the expression of caspase-3 was lower in the hemin-IR group in comparison to those of the IR group. The results in the ZnPP-IR group were just the opposite to those of the hemin-IR group.

CONCLUSIONHO might play a protective role in hepatic IR injury in rats, and this effect may be related to the inhibition of hepatocellular apoptosis.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Heme Oxygenase (Decyclizing) ; metabolism ; Hepatocytes ; metabolism ; pathology ; Liver Diseases ; metabolism ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Protoporphyrins ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology