OBJECTIVETo investigate the influence of therapeutic bloodletting at Jing-well points and hypothermia on acute cerebral edema after traumatic brain injury (TBI) in rats.

METHODSSeventy-five SD rats were randomly divided into sham-operation group (Sham), TBI group (TBI), bloodletting group (BL), mild-induced hypothermia group (MIH), and bloodletting plus MIH group (BL + MIH) (n = 15). The model of TBI was established by electric controlled cortical impactor (eCCI). The rats of BL group were bloodletting at Jing-well points immediately after injury, twice daily. While the MIH group was settled on a hypothermia blanket promptly after TBI for 6 hours, so that the temperature dropped to 32 degrees. Each of measurement was performed after 48 hours. Magnetic resonance imaging (MRI) was used to evaluate the dynamic impairment of cerebral edema after TBI (n = 3). In addition, mNSS score, measurements of wet and dry brain weight, and Evans Blue assay were performed to investigate the neurologic deficit, cerebral water content (n = 8), and blood-brain barrier permeability (BBB), (n = 4), respectively.

RESULTSMRI analysis showed that the cerebral edema, hematoma and midline shifting of rats in TBI group was more serious than other treatment group. Meanwhile compared with TBI group, the mNSS scores of every treatment group were meaningfully lower (all P < 0.05). Furthermore, treatment with BL+ MIH group was superior to the separated BL and MIH group (all P < 0.01). In addition, brain water content of each intervention group reduced to varying degrees (all P < 0.05), especially that of MIH group and BL + MIH group (P <0.01). BBB permeability of each treatment group was also significantly improved (all P < 0.01), and the improvement in MIH group and BL + MIH group was much better than the BL alone group (P < 0.05, P < 0.01).

CONCLUSIONOur major finding is that bloodletting at Jing-well points and MIH can reduce cerebral edema and BBB dysfunction and exert neuroprotective effects after TBI. The results suggest that the combination of BL and MIH is more effective than other treatment being used alone.

Animals ; Blood-Brain Barrier ; Bloodletting ; Brain ; pathology ; Brain Edema ; prevention & control ; Brain Injuries ; therapy ; Hypothermia, Induced ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo simulate the chemical microenvironment of injured brain tissue, and to explore the effect of this chemical microenvironment on temperature sensitive umbilical cord mesenchymal stem cells (tsUC).

METHODSRat models of traumatic brain injury (TBI) were made by fluid percussion injury, and then the brain tissue extracts of the injured regions were acquired. Human umbilical cord mesenchymal stem cells (UC) were isolated and cultured, and the tsUC were obtained through the infection of temperature-sensitive Simian 40 Large T- antigen (ts-SV40LT) retrovirus. After that, both the two kinds of cells were cultured on the polyacrylamide gels which mimicking the elastic modulus of brain. Four groups were included: UC cultured under normal temperature (UC group), UC cultured added brain tissue extract under normal temperature (UC plus extract group), tsUC cultured under mild hypothermia (tsUC group), and tsUC added brain tissue extract under mild hypothermia for 3 days, then normal temperature for 4 days (tsUC plus extract group). After 24 hours, the apoptosis level was checked. Cell growth and morphological changes in each group were given dynamic observation. Seven days later, cell immunofluorescences were implemented for examining neural differentiation level.

RESULTSCompared with UC plus extract group, the apoptosis and proliferation in UC plus extract group were significantly reduced (P < 0.01) and increased (P < 0.01) respectively. Cell immunofluorescence showed that the both GFAP and Neuron positive cells were significantly enhanced in UC plus extract group than those in tsUC plus extract group.

CONCLUSIONtsUC combining with mild hypothermia could significantly reverse injury induced cell apoptosis, improve cell proliferation and neural differentiation under chemical microenvironment after brain injury, which confirmed the adaptation and resistance of tsUC under mild hypothermia after TBI.

Animals ; Apoptosis ; Brain ; cytology ; pathology ; Brain Injuries ; pathology ; Cell Proliferation ; Humans ; Mesenchymal Stromal Cells ; chemistry ; Neurons ; cytology ; Rats ; Temperature ; Umbilical Cord ; cytology

Objective To establish the immortalized umbilical cord mesenchymal stem cells(UCMSCs) mediated by human telomerase reverse transcriptase (hTERT) so as to offer enough UCMSCs for in vitro or clinical researches. Methods Human UCMSCs were isolated and cultured to passage 4,and then identified by flow cytometry analysis. The pLXSN-hTERT plasmid was transfected into UCMSCs by liposome to establish the immortalized UCMSCs. Expressions of hTERT mRNA and protein were respectively tested by RT-PCR and immunocytochemistry. The cell cycle kinetics of the passage of hTERT-UCMSCs and UCMSCs were detected with flow cytometry. Results Flow cytometry analysis showed that UCMSCs expressed CD29, CD44 and CD 105 strongly, but CD31, CD34 and CD45 slightly; these were right the features of human UCMSCs. RT-PCR showed that hTERT mRNA was expressed low in UCMSCs, and high in pLXSN-hTERT transfected UCMSCs.Immunocytochemistry revealed that fluorescence intensity of cell nuclei was increased significantly after transfection and that hTERT was expressed all in cell nuclei. Flow cytometry analysis suggested that the number of hTERT-UCMSCs at phase S was increased significantly, and the cells would be able to passage stably (more than 35 passages). Conclusions The immortalized UCMSCs can be established by hTERT transfection, and the number of immortalized UCMSCs can meet the need of in vitro or clinical researches.

OBJECTIVETo study whether antisense oligonucleotides and ultrasonic microbubble intensifier transfection combined with ultrasound irradiation is an effective and directional way in reversing multidrug resistance (MDR) in tumors.

METHODSMdr1, mrp, and lrp genes antisense oligonucleotides on the ultrasound microbubble intensifier were transfected for the human HepG2/ADM cell lines and then the cells were radiated with low intensity ultrasound. The effects of the reversion of carcinoma cells' MDR and the reduction of their malignancy and growth capability in vitro and in vivo were assessed using RT-PCR, Western blot and MTT.

RESULTSThe treatment restrained the multiplication of the human HepG2/AMD cell lines. The levels of their mRNA and protein of cells' mdr1 and mrp genes dropped significantly. Growth of the subcutaneous transplanted tumors in the nude mice decreased.

CONCLUSIONSTransfection of MDR genes antisense oligonucleotides on the ultrasonic microbubble intensifier combined with low intensity ultrasound radiation may serve as a new treatment method for hepatocellular carcinoma.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; Liver Neoplasms ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Microbubbles ; Oligonucleotides, Antisense ; genetics ; Transfection ; Ultrasonics

Cardiovascular Diseases ; Gastrointestinal Microbiome ; Humans ; Methylamines

Objective:To explore the composition characteristics of rhizosphere soil under Rehmannia glutinosa-Zea mays intercropping model，and screen out special signal substances in rhizosphere soil of R. glutinosa under intercropping Z. mays， so as to provide the basis for the study of allelopathic substances in continuous cropping obstacle of R. glutinosa. Method:In this experiment，rhizosphere soils of R. glutinosa under Z. mays intercropping and R. glutinosa single cropping models in July，August，September and October were taken as the research objects， and the volatile organic compounds in ethyl acetate fraction were analyzed by gas chromatography-mass spectrometry （GC-MS）. Principal component analysis （PCA）， hierachical cluster analysis （HCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） analysis were performed on the data by SIMCA 14.1 to screen out potential differences in volatile organic compounds between the two models. Result:The types of volatile organic compounds in intercropping and single cropping models were mainly hydrocarbons， alcohols， esters， ketones， amides， acids and other substances. Specifically， the average relative contents of hydrocarbons，esters and amides in intercropping model were 58.46%，32.15% and 5.42% respectively，while the relative contents of hydrocarbons，esters and amides in single cropping model were 37.27%，36.11% and 21.13%. The results of PCA and HCA showed that the characteristics of volatile organic compounds in the ethyl acetate fraction of rhizosphere soil under intercropping and single cropping models could be clearly divided into two categories，the screening results of potential differential components based on OPLS-DA analysis indicated that various components， such as dibutyl phthalate，（Z）-9-oleamide，β-caryophyllene，dioctyl iso-phthalate， phthalate （2-propylamyl） diester， n-hexadecane，octodecane， n-heneicosane， were screened from rhizosphere soil under the two models. Conclusion:The R. glutinosa-Z. mays intercropping model has certain effects on the volatile organic compounds in the rhizosphere soil of R. glutinosa，and the effect of the selected components on the growth and quality characteristics of R. glutinosa still need to be further studied.

Coal miners are a high-risk occupational group of work-related musculoskeletal disorders(WMSDs). This disease not only causes a reduction in the quality of life of workers, loss work ability, and decline in work efficiency, but also becomes an important factor leading to increase social medical burden. At present, most of the evaluation of WMSDs of coal miners at home and abroad adopts the international general scale. Chinese scholars have developed a WMSDs questionnaire that meets the occupational characteristics of Chinese population based on the international general scale—the Chinese Musculoskeletal Disorders Questionnaire. The questionnaire has good reliability and validity, and is a reliable and effective tool for evaluating WMSDs. The prevalence of WMSDs in coal miners is relatively high. The occupational factors that affect WMSDs in coal miners are mainly physical load, postural load, mental load, work organization, and work environment. The individual factors are mainly age, working experience, and education level. It′s recommended to prevent and control the occurrence of coal miners′ WMSDs through multi-level intervention measures such as strengthening education, changing the working posture, and improving the working conditions of coal miners, and rationally arranging labor organizations.

Objective: To discuss the phenotypic character and the HPLC fingerprints of radial striations from different germplasms Rehmanniae Radix. Method: The changes in the shape and column diameter of the radial striations of Rehmanniae Radix were observed and measured in the whole growth period. Besides,the HPLC fingerprints of the root,radial and un-radial striations were established to sign the chemical quality and analyzed by principal component analysis(PCA)and systematic cluster analysis. Result: There were significantly differences and regularities in the shape and proportion of the radial striations of different germplasms Rehmanniae Radix. The fingerprints showed the consistency between different types of chemical ingredients,and the differences in chemical quality characteristics mainly lay in the content of chemical compositions and theirs relative ratio. The results of PCA indicated that active ingredients, such as acteoside,catalpol,rehmaionoside D,rehmaionoside A and leonuride, were involved in the quality expression of different parts from various germplasms of Rehmanniae Radix,but each ingredient had a distinctive contribution rate to the differential quality expression between different parts from various germplasms of Rehmanniae Radix. However,the other components involved in the differential quality expression had different contribution rates in different germplasms.The systematic cluster analysis indicated that great differences in the chemical quality between the radial striations and un-radial striations of Beijing-1,Qinhuai,Qinhuai Zheng and 1706 germplasms,but with small differences in 85-5 and Baixuan germplasms. Conclusion: There are differences in phenotypic character of the radial striations and HPLC fingerprints between different germplasms Rehmanniae Radix.

OBJECTIVES: To study the transcriptomic changes of astrocytes in the brain of rats exposed to methamphetamine (METH) and its possible mechanism in neurotoxicity. METHODS: The rats were intraperitoneally injected with METH (15 mg/kg) every 12 h for 8 times in total to establish the subacute rat model of METH. After the model was successfully established, the striatum was extracted, and astrocytes were separated by the magnetic bead method. Transcriptome sequencing was performed on selected astrocytes, and the differentially expressed genes were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. RESULTS: A total of 876 differentially expressed genes were obtained by transcriptome sequencing, including 321 up-regulated genes and 555 down-regulated genes. GO analysis revealed that differentially expressed genes were mainly concentrated in cell structure, biological process regulation, extracellular matrix and organelle functions. KEGG pathway enrichment analysis showed that steroids biosynthesis, fatty acid biosynthesis, peroxisome proliferators-activated receptor (PPAR), adenosine 5'-monophosphate-activated protein kinase (AMPK) and other signaling pathways were significantly changed. CONCLUSIONS METH can cause structural changes of astrocytes through multiple targets, among which cellular structure, steroids biosynthesis and fatty acid biosynthesis may play an important role in nerve injury, providing a new idea for forensic identification of METH related death.
Animals ; Astrocytes ; Brain ; Gene Expression Profiling ; Methamphetamine/pharmacology* ; Rats ; Signal Transduction ; Transcriptome

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires