OBJECTIVETo investigate the antibacterial activity of decoction of Radix glycyrrhizae against Streptococcus mutans (S. mutans) in vitro.

METHODSThe decoction of Radix glycyrrhizae was prepared by boiling particles of Radix glycyrrhizae, the diameter was 0.2-3.2 mm. In distilled water and filtered, the filtrate was collected for study. The minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC) of the decoction against S. mutans were detected using double dilution. The effect of decoction on growth and acidogenic profile of S. mutans were investigated by detecting the Abs of bacteria suspension and the pH value of medium at definite time intervals(0, 3, 7, 12, 23, 40 h) during cultured.

RESULTSThe MIC determined for decoction was 50 mg x mL(-1) and there was no bactericidal effect when concentration of decoction lower than 100 mg x mL(-1). The decoction inhibitted multiplication of bacteria significantly and the effects became stronger with concentration increasing. The decoction also inhibitted S. mutans producing acid and the effect became stronger with concentration increasing. The most efficient inhibition were observed when incubated 12 hours.

CONCLUSIONThe decoction of Radix glycyrrhizae can inhibite the growth and acid-production of S. mutans in vitro.

Anti-Bacterial Agents ; Bacteria ; In Vitro Techniques ; Microbial Sensitivity Tests ; Plant Extracts ; Streptococcus mutans

OBJECTIVETo establish an efficient and stable method for protein extraction of Streptococcus mutans.

METHODSThe collected bacteria were treated by freeze-thaw and ultrasonic (method 1), ultrasonic (method 2), boiling (method 3), boiling and ultrasonic (method 4), respectively. The index such as state of bacteria broken, concentration of extracted protein and SDS-PAGE of protein were employed to evaluate the effects of above four methods.

RESULTSBeside the method 3, the other three methods could break the bacteria effectively, of which ultrasonic was the key factor. The pattern of SDS-PAGE which treated by method 1, method 2 and method 4 was almost same, but method 1 resulted in the best abundance. There was significantly difference among the four protein concentration extracted by four methods (P < 0.05). All methods exhibited good stability and reproducibility.

CONCLUSIONMethod of freeze-thaw and ultrasonic resulted in an efficient proteins extraction of Streptococcus mutans which demonstrated good stability and reproducibility and easy to handle.

Bacterial Proteins ; Reproducibility of Results ; Streptococcus mutans

OBJECTIVETo explore the expansion method of high purity NK cells from human peripheral blood and explore the changes in biological functions of NK cells after ex vivo expansion.

METHODSNK cells were isolated from peripheral blood mononuclear cells (PBMNCs) by using miniMACS (Magnetic cell-selection) and NK Cell Isolation Kit II, and cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of interleukin (IL)-2 and/or IL-12, IL-15 for 15 days. Cultures were semi-exchanged with fresh media and cytokines every 3 days. Evaluation for cell expansion, phenotype, perforin and granzyme B mRNA expressions, and IFN-gamma secretion before and after the culture period.

RESULTSCD3(-) CD56(+) cells concentration increased from (11.2 +/- 5.2)% to (94.2 +/- 3.5)%. In group IL-2 + IL-15 and IL-2 + IL-15 + IL-12 group, cells were expanded 50.5 +/- 4.3 and 52.3 +/- 6.7 - fold, respectively, being significantly higher than that in other three groups [(15.4 +/- 1.1 fold in IL-2 group, 19.9 +/- 3.9 fold in IL-2 + IL-12 group, 6.1 +/- 1.0 fold in control group)] (P<0.01), but no significant difference between each other (P>0.05). The purity of CD3(-) CD56(+) NK cells was over 94% in all groups except the control. The perforin and granzyme B mRNA expressions of expanded NK cells in four experimental groups were significantly higher than those of before expansion (P<0.01) and the expressions in IL-2 + IL-15 and in IL-2 + IL-12 + IL-15 group were significant higher than in other three groups (P<0.01) while no significant difference between each other (P>0.05). IFN-gamma levels in the supernatants of four experiment groups were significantly higher than that in control group (P<0.01) and its levels order was IL-2 + IL-15 + IL-12 group > IL-2 + IL-12 group > IL-2 + IL-15 group > IL-2 group (P<0.01).

CONCLUSIONHigh purity NK cells isolated by negative selection using miniMACS can be efficiently expanded with IL-2 + IL-15, and their biological functions were enhanced.

Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; pharmacology ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukins ; pharmacology ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; metabolism ; Perforin ; metabolism

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype soft ware were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient. The results showed that donor chimerism was similar by the two methods. The median number of informative alleles was 6.3 (4 - 9) by Profiler Plus and 4.9 (2 - 6) by Cofiler Plus. The donor alleles appeared in 26 patients on day 28 post transplantation. One patient was not observed to appear donor alleles. 14 patients with 100% donor chimerism (DC) had stable engraftment and they still survive in free leukemia. 9 patients had unstable mixed chimerism (DC: 0% - 90.2%), and 5 of them relapsed after allo-HSCT, 6 patients died. Decrease of donor chimerism appeared prior to graft rejection and disease relapse. The incidence of GVHD was higher in group of full donor chimerism. It is concluded that dynamic monitoring donor chimerism by STR-PCR in combination with all auto-capillary electrophoresis is a valuable tool for predicting graft rejection, disease relapse and occurrence of GVHD, and provides a basis for early clinical intervention in the patients received allo-HSCT.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; prevention & control ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Peripheral Blood Stem Cell Transplantation ; methods ; Polymerase Chain Reaction ; methods ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous

This study was purposed to explore the changes in biological functions of human peripheral blood derived NK Cells after ex vivo expansion with different combinations of interleukin IL2 and/or IL12, IL15. According to different combination of cytokines, cultured NK cells were divided into 4 groups: group IL2, group IL2 + IL12, group IL2 + IL15 and group IL2 + IL15 + IL12. The group in which NK cells were cultured without cytokines was used as control. The cytotoxicity of cultured NK cells to target K562 cells was determined by using cell counting kit-8; the level of IFN-gamma in supernatants of NK cell culture was detected by ELISA; the perforin and granzyme B mRNA expressions were assayed by competitive quantitative RT-PCR. The results showed that the cytotoxicity of expanded NK cells in groups cultured with cytokines at different E:T ratio was significantly higher than that in group without cytokines (p < 0.01), although the cytotoxicity of NK cells in IL2 + IL15 + IL12 group seem to be slightly higher than that in IL2 + IL15 group, but there was no statistic difference (p > 0.05). The IFN-gamma levels in the supernatants of NK cell culture in the presence of cytokines significantly increased, and the IFN-gamma levels in IL2 + IL15 + IL12 group and IL2 + IL12 group were significantly higher than that in others (p < 0.01). The expressions of perforin and granzyme B mRNA of expanded NK cells in groups cultured with cytokines was significantly higher than that in control group (p < 0.01), and was consistent with cytotoxicity of NK cells. It is concluded that there are differences in the functions of NK cells cultured with different cytokines. IL2 and IL15 have synergistic effect on strengthening cytotoxicity of NK cells and promoting cell expansion. However, the main function of IL12 promotes NK cells to secrete IFN-gamma, which plays a role in immunoregulation.
Humans ; Interferon-gamma ; secretion ; Interleukin-12 ; administration & dosage ; pharmacology ; Interleukin-15 ; administration & dosage ; pharmacology ; Interleukin-2 ; administration & dosage ; pharmacology ; K562 Cells ; Killer Cells, Natural ; drug effects ; immunology

Objective To explore the short-term and medium-term efficacy of modified loop plate suspension fixation in the treatment of acromioclavicular joint dislocation.Methods A retrospective analysis was performed in 72 patients with acromioclavicular joint dislocation and treated in Yangzhou Hospital of Traditional Chinese Medicine from December 2020 to December 2022.The patients were divided into modified group(n=37)and traditional group(n=35)according to different surgical methods.The modified group was treated with modified loop plate suspension fixation,and the traditional group was treated with clavicular hook plate fixation.The perioperative indexes,Constant-Murley score,and shoulder joint mobility and pain visual analogue scale(VAS)score before operation,3 months and 6 months after operation,postoperative complications rates,excellent and good rate of shoulder joint function recovery at 6 months after operation were compared between the two groups.Results The incision length and recovery time in the modified group were significantly shorter than those in the traditional group(P＜0.001),and the intraoperative blood loss was significantly less than that in the traditional group(P＜0.001).The Constant-Murley score,shoulder flexion and abduction activity in both groups at 3 months and 6 months after operation were significantly higher than before operation(P＜0.05),and these improvements were greater in the modified group(P＜0.05).The VAS scores of the two groups at 3 months and 6 months after operation were significantly lower than before operation(P＜0.05),and these decreases were greater in the improved group(P＜0.05).There were no significant differences in the excellent and good rate of shoulder joint function recovery and the incidence of postoperative complications between the two groups.Conclusions The modified titanium loop plate suspension fixation in the treatment of acromioclavicular joint dislocation can reduce surgical trauma and postoperative pain,and promote the recovery of shoulder joint function.

To demonstrate the phylogenetic evolution, the molecular characteristics of the motif of HA protein cleavage site and the varieties at the receptor binding sites of the hemagglutinin gene of the duck-origin H9N2 subtype avian influenza viruses, sequence alignment and phylogenetic analysis were performed by MEGA 4.1 Neighbor-Joining method.. The results revealed that the duck-origin H9N2 AIV viruses originated from CK/BJ/1/94-like and North-Ame-like, all the duck-origin H9N2 AIV viruses from mainland China belonged to CK/BJ/1/94-like and formed multiple genotypes through complicated re-assortment, while other duck-origin H9N2 AIV, isolated from other countries in Aisa, American and European such as Korea, Japan, Alberta, Austria, Switzerland, Iran, belonged to the North-Ame-like phylogenetic lineage. The amino acids at positions 183, 190, and 226 of the receptor binding sites of North-Ame-like group isolates had highly conserved H, E and Q respectively. In contrast with duck-origin H9N2 AIV viruses isolates from mainland China, the amino acids had N at positions 183, A, T, or V at 190, L or Q at 226, which was the same as the chicken-origin H9N2 AIV from mainland China. Most newly isolated chicken-origin H9N2 AIV in Fujian Province in Southern China had L at position 226 emphasized the higher risk of cross-infection between the chicken-origin and duck-origin H9N2 AIV in China.
Animals ; China ; Ducks ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; Influenza A Virus, H9N2 Subtype ; chemistry ; classification ; genetics ; isolation & purification ; Influenza A virus ; chemistry ; classification ; genetics ; Influenza in Birds ; virology ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; virology ; Sequence Alignment

OBJECTIVETo investigate the oral health knowledge and habits of freshmen of Sichuan University and promote the prophylactic therapy of dental diseases in the university.

METHODSThe table and the standard on oral health survey authorized by WHO were employed in this study.

RESULTSOnly 13.1% of all the investigated freshmen had periodic oral health examination. 74.4% brush teeth twice per day, only 7.7% brush teeth over 3 minutes, 16.6% insist in fluoridated toothpaste, and only 6% had applied dental floss. 51.9% do not often take snack food and 94.7% are non-smokers; (4) Tests of prophylactic therapy of dental caries and periodontal diseases and oral health knowledge have poor results.

CONCLUSIONThe freshmen have limited knowledge of oral health and lack serious attitude toward oral cleaning behaviors.

Health Knowledge, Attitudes, Practice ; Humans ; Oral Health ; statistics & numerical data ; Oral Hygiene ; statistics & numerical data ; Students ; Universities

OBJECTIVETo detect the expression of B7-H1 gene in bone marrow mononuclear cells (BMMNCs) from leukemia patients and explore its clinical implications.

METHODSThe B7-H1 mRNA expression levels of BMMNCs from 74 newly diagnosed leukemia patients and 10 normal volunteers were detected by real-time quantitative PCR. At the same time, BMMNCs from 12 patients in complete remission (CR) after chemotherapy and 5 in relapse were followed up. The correlation between the clinical features of 74 de novo leukemia patients and the expression level of B7-H1 gene was analyzed.

RESULTSThe mRNA expression level of B7-H1 gene in BMMNCs from de novo leukemia patients (RQ = 0.125) was lower than that from normal control (RQ=1). When patients achieved CR the gene expression level (RQ = 69.07) was significantly higher than that before CR (P = 0.001). After relapsed, its level (RQ=4) was still higher than that before CR (P > 0.05). No clinical parameters such as gender, age, peripheral white blood count, blast cells ratio in BM, CD34 positive cells were significantly correlated with the expression level of B7-H1 except the response to therapy. The initial expression level of B7-H1 gene in non CR patients after therapy was significantly higher than that in CR patients (RQ = 26. 91, P = 0.005).

CONCLUSIONThe mRNA expression level of B7-H1 gene in newly diagnosed leukemia patients is lower than that in normal controls, and is higher in CR patients than in newly diagnosed patients. There is a correlation between the gene expression level and responsiveness to therapy.

Adult ; Antigens, CD ; metabolism ; B7-H1 Antigen ; Female ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

The aim of this study is to investigate the effect of echinacoside (ECH) on cholinergic neurotransmitter extracellular of hippocampus and striatum and its possible mechanisms of neuro-protective effect against vascular dementia rats. In this study brain microdialysis technique combined with HPLC-IMER-ECD (high-performance liquid chromatography-immobilized enzyme reactor-electrochemical detector) was used. The bilateral common carotid arteries occluded in two times operation at 72 h interval for vascular dementia model rats were used and the successful vascular dementia model rats were examined by Morris water maze. The content of acetylcholine (ACh) and choline (Ch) of microdialysate extracellular of hippocampus and striatum was determined by HPLC-IMER-ECD and the AChE activity in the hippocampus was measured. The results showed that the success rate of vascular dementia model was 83.08% after six weeks; the results also showed that echinacoside and galantamine could increase the content of ACh and reduce the content of Ch extracellular of hippocampus and striatum significantly and the AChE activity increased significantly compared with that of the model group. The results suggested that echinacoside could promote the recovery of cholinergic neurotransmitter levels in vascular dementia rats' brain, which may be one of the mechanisms of neuro-protection.
Acetylcholine ; metabolism ; Animals ; Choline ; metabolism ; Corpus Striatum ; metabolism ; Dementia, Vascular ; metabolism ; Glycosides ; pharmacology ; Hippocampus ; metabolism ; Male ; Neuroprotective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley