BACKGROUND:Needle-knife release of the ligamentum flavum can effectively improve symptoms in patients with lumbar degeneration,and ultrasound guidance can increase the precision of needle-knife release;however,the specific effects of needle-knife release of the ligamentum flavum on the degenerated intervertebral discs and the possible mechanisms remain to be clarified. OBJECTIVE:To investigate the effect of ultrasound-guided needle-knife release of the ligamentum flavum. METHODS:Twenty-four New Zealand rabbits were randomized into control(n=6)and model(n=18)groups.A rabbit model of lumbar disc degeneration model was established in the model group by cutting the supraspinous and interspinous ligaments of the L5/6 and L6/7 segments to maintain a standing posture and apply axial load to the lumbar spine.After successful modeling,the model rabbits were subdivided into a control group,a model group,an ultrasonic needle-knife group,and a sham needle-knife group according to a random number table method,with six animals in each group.The ultrasonic needle-knife group underwent ultrasound-guided needle-knife release of the right yellow ligament of L7/S1,once every week,for a total of four times.The needle-knife approach in the sham needle-knife group was the same as that in the ultrasound needle-knife group,but the ligamentum flavum was not released.At 30 days after the intervention,MRI was used to observe the changes in the signal intensity of the nucleus pulposus within the L7/S1 segment.Hematoxylin-eosin staining was used to observe the morphological changes of the L7/S1 segment.Immunohistochemical staining was used to detect the expression of type I and II collagen in the nucleus pulposus of the L7/S1 segment.RT-PCR and western blot were used to detect the expression of integrin α5 and β1,p38,and nuclear factor κB in the L7/S1 segment. RESULTS AND CONCLUSION:MRI findings indicated that the nucleus pulposus of the intervertebral disc of rabbits in the model group was gray-black in color,and the gray value of the nucleus pulposus was significantly lower than that of the control group(P<0.01).The brightness of the nucleus pulposus of the intervertebral disc of the rabbits in the ultrasonic needle-knife group was elevated compared with that of the model group,and the gray value of the nucleus pulposus was higher than that of the model group(P<0.01).Results from hematoxylin-eosin staining showed that in the model group,the shape of the nucleus pulposus was irregular,the number of nucleus pulposus cells was reduced,the extracellular matrix was compressed,the fibrous ring was ruptured,the structure and boundary of the end plate were unclear,and the chondrocytes were arranged disorderly.Compared with the model group,the ultrasonic needle-knife group showed an increase in the number of the nucleus pulposus,an improvement in the rupture of the fibrous ring,and more regular arrangement of cartilage endplate cells.Results from immunohistochemical staining showed an increase in positive expression of type I collagen(P<0.01)and a decrease in positive expression of type II collagen in the nucleus pulposus of the model group compared with the control group as well as a decrease in positive expression of type I collagen and an increase in positive expression of type II collagen in the nucleus pulposus of the ultrasonic needle-knife group compared with the model group(P<0.01).RT-PCR and western blot assays showed that the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the model group were increased compared with that in the control group(P<0.01);the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the ultrasonic needle-knife group was decreased compared with that in the model group(P<0.01).To conclude,ultrasound-guided needle-knife release of the ligamentum flavum can improve the degree of lumbar disc degeneration in rabbits,which may be related to the inhibition of p38 and nuclear factor-κB expression by modulating integrin α5 and β1 expression.

In this research,Microtox technology was used to evaluate the comprehensive toxicity of Shen-Fu injection,which was an exclusive product of the pharmaceutical company.Firstly,vibrio fischeri was used as test strain.The best test system and methodology were identified.Under the best test conditions,vibrio fischeri was firstly used in the luminescent bacteria comprehensive toxicity of the exclusive product Shen-Fu injection.The results showed that in the 2 mL reaction system,the best recovery liquid volume was 0.9 mL/ freeze-dried vial,50 μL bacterial suspensions/ sample,the detection time was 10 min after adding bacterial suspensions,the pH was 5-10,the luminous intensity was 800 000-12 000 000 for 10 rin.The RSD of replication experiment and intermediate precision test was ? and ?,respectively,which fitted to the requirements.There was no significant difference on EC50 values among 9 batches of tested Shen-Fu injection (41.07％,RSD ＜ 5％).It was concluded that there was significant concentration-effect relationship between Shen-Fu injection and the toxicity of vibrio fischeri.There was no significant difference on EC50 values among different batches.It indicated that there was small quality volatility among different batches of Shen-Fu injection.The control on product manufacturing was strong.

Microtox technology is one of quick toxicity test methods for determining the poisonous or toxic substances in the environment by luminous bacteria as a indicator organism.This paper reviewed the appearance and development of microtox technology.Besides,we expounded the methods and stepwise technique map and addressed key questions of microtox technology for the evaluation of the comprehensive toxicity of Chinese medicinal materials.In conclusion,microtox technology is a promising method from a vantage point of the toxicity detection of Chinese medicinal materials.

This study aimed at exploring the application of microtox technology to the evaluation of comprehensive toxicity of Sheng Mai injections.Characteristic parameters of vibrio fischeri (CS234) were measured by methodology inspection under different conditions to optimize the reaction condition with reliable technology.It was the first experiment to accomplish the comprehensive toxicity of Sheng Mai injections from various manufacturers using vibrio fischeri under the target condition.It was found that parameters of the optimum condition contained 0.9 mL recovery liquid volume in each freeze-dried vial and 50 μ L bacteria suspension of each sample,being included in 2 mL solution in aggregate under 10 mins' detection time after adding bacteria suspension with the favorable pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million within 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 22.10％,34.10％ and 46.04％,respectively,presenting significant statistical differences (P＜0.05).These results demonstrated that the growth of biological detection standards of Sheng Mai injection was highlighted a long way to go.Besides,the microtox-based fast test for the evaluation of the comprehensive toxicity of Sheng Mai injection showed a prospective application to controlling the quality of different products from manufacturers.

In this research,we chiefly explored a new fast test method--microtox technology to evaluate the comprehensive toxicity of Shen Mai injection.We investigated characteristic parameters of vibrio fischeri (CS234) under different conditions in the pursuit of the optimum test parameters of the reliable method;and then locked the best parameters for Shen Mai injection assay with the products from different manufacturers by microtox fast test.As a result,the optimum reaction condition included 0.9 mL recovery liquid in each freeze-dried vial and 50 μL bacteria suspension of each sample within 2 mL solution in aggregate with 10-mins detection after adding bacteria suspension which pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million in 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 35.60％,92.34％ and 146.57％,differing among the samples from three representative drug manufacturers,respectively (P ＜ 0.01).In conclusion,the concentration-effect relationship of Shen Mai injection existed in the toxicity assessment of vibrio fischeri with various ECs0 values from the three manufacturers.These results suggested that biological detection standards of Shen Mai injection presented great growth potential.Microtox-based fast test in the evaluation of comprehensive toxicity of ShenMai injection showed favorable application prospects over controlling the quality of different sources of products.

Objective To observe the efficacy of urinary kallidinogenase plus batroxobin in the treatment of acute cerebral infarction.Methods 105 patients with acute cerebral infarction were selected and divided into 3 groups:urinary kallidinogenase group (35 cases),combined treatment group (39 cases),batroxobin group (31 cases).The NIHSS score,Barthel Index,MRS score were evaluated before treatment and 10 days after treatment,and the clinical efficacy was compared.Results The NIHSS score significantly decreased after treatment in the three groups (all P ＜ 0.05).The effective rate of combined treatment group was better than that of the single treatment group (urinary kallidinogenase group or batroxobin group) [79.5 ％ (31/39),71.4 ％ (25/35),35.4％ (11/31) (x2 =15.801,P =0.001)].The Barthel Index and MRS score of combined treatmentgroup were better than those of the batroxobin group (P =0.003),not better than the urinary kallidinogenase group (P =0.766).Conclusion Urinary kallidinogenase plus batroxobin is effective in the treatment of acute cerebral infarction.

Adult ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Appendectomy ; Appendiceal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Appendicitis ; etiology ; surgery ; Diagnosis, Differential ; Female ; Granular Cell Tumor ; complications ; metabolism ; pathology ; surgery ; Humans ; Paraganglioma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism

BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research.
OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s.
METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by
immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein.
RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.

Circulating tumor cells (CTCs) are present in the peripheral blood,and play an important role in the recurrence and metastasis of lung cancer.As the researches about CTCs and targeted therapy move along,it is found that CTCs can be used as a liquid tumor tissue to guide clinical treatment and have a certain significance in the clinical staging and prognosis evaluation of lung cancer.

OBJECTIVETo comparative study the acute toxicity of four extracts from Xanthii Fructus in mice.

METHODObserved the toxic manifestations in mice which were given the four extracts by intragastric administration and calculated the LD50 of the four extracts from Xanthii Fructus.

RESULTThe toxic manifestations of the mice given water extract by intragastric administration were repose, pilo-erection, cyanosis, intention tremor, respiratory inhibition, loss of righting reflex and convulsion . The toxic manifestations of the mice given ethanol extract by intragastric administration were repose, abdominal respiration, intention tremor, intermittent convulsions, incontinence. The LD50 of Xanthii Fructus processed water extract, processed ethanol extract, crude water extract, crude ethanol extract were material drug 155.93, 317.80, 167.6, 275.41 g x kg(-1), respectively.

CONCLUSIONThe acute toxicity of water extract is distinctly stronger than that of ethanol extract, but there is no marked distinguish between crude and processed extract.

Animals ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Female ; Fruit ; chemistry ; Lethal Dose 50 ; Male ; Mice ; Reflex, Righting ; drug effects ; Respiration ; drug effects ; Xanthium ; chemistry