To developing a simple, rapid and sensitive immunoaffinity method for extracting DNA before amplification by the polymerase chain reaction(PCR). Using monoclonal anti-DNA bound to colloidal-gold beads(which called immunocolloidal-gold ) and extracting DNA from the specimen before amplification by PCR. The immunoaffinity method we use is extremely efficient in extracting DNA at low concentration, also, the beads carrying the DNA/anti-DNA complexes can be added directly to a PCR mixture without elution, and the presence of PCR inhibitors in specimens has no effect on amplification. When applied to serial dilution bacteria suspension, the detection of limit can be 100 CFU/mL, and the sensitivity of detecting artificial soil sample and milk sample were 101 and 100 CFU/mL. The immunoaffinity method described here can sever as a sensitive and practicable method for extracting DNA , particularly where samples have low concentrations of DNA or where the material is in poor condition.

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism

OBJECTIVETo study the integrin beta1 mRNA changes after orthodontic tooth movement in normal teeth and periodontitis teeth of rats.

METHODSThe OD of positively stained osteoclasts for integrin beta1 mRNA using in situ hybridzation was detected after orthodontic tooth movement in normal teeth and periodontitis teeth groups.

RESULTSIntegrin beta1 mRNA expression were detected on all osteoclasts in tooth movement samples of normal and periodontitis teeth. There were stronger positive signals after given orthodontic force in both of the two groups. But no differences were found after 0.5, 1, 2, 3, 5, 7, 10 days since orthodontic tooth movement. The integrin beta1 mRNA signals in normal tooth movement group were not different from that in periodontitis group.

CONCLUSIONThe integrin beta1 of osteoclasts may play a role in the stability and remodeling of periodontal ligament in orthodontic tooth movement. There were no difference in the OD of integrin beta1 mRNA staining in orthodontic tooth movement between normal teeth group and periodontitis teeth group.

Animals ; Integrin beta1 ; metabolism ; Osteoclasts ; Periodontal Ligament ; Periodontitis ; physiopathology ; RNA, Messenger ; metabolism ; Rats ; Tooth Movement Techniques

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

Liver is one of the most important organs in the human body. Liver diseases are also a major threat to human health and longevity. Hepatic decompensation treatment is quite difficult due to multiple reasons. Extracorporeal liver support devices are unable to solve this problem, and there is a severe shortage of orthotopic liver transplant donors. Study of pluripotent stem cell-derived hepatocytes and organoids can determine not only hepatocyte fate, but also liver development, regeneration mechanisms, and pathophysiology. Furthermore, it can be used for drug screening in order to provide a stable source of functional hepatocytes for future transplantation therapy. Culture of pluripotent stem cell-derived hepatocytes and organoids has a self-organizing process similar to liver development, i.e., starting with changes in several key factors, and eventually forming functionally complex cells/organs. This paper introduces the main methods and progress of pluripotent stem cell-derived hepatocytes and organoids, with hope to provide clues for future research.
Cell Differentiation ; Hepatocytes ; Humans ; Induced Pluripotent Stem Cells ; Liver ; Organoids ; Pluripotent Stem Cells

OBJECTIVETo study the integrin beta3 mRNA changes after orthodontic treatment on normal teeth and periodontitis teeth in rats.

METHODS96 adult SD rats of 10 weeks old were randomly divided into normal tooth move-ment group and periodontitis tooth movement group. The rats in the two groups were sacrificed after 0 d, 12 h, 1 d, 3 d, 5 d and 7 d of tooth movement. The alveolar specimens were prepared. The integrin beta3 mRNA were detected using in situ hybridization in the specimens. The OD index of positively stained osteoclasts for integrin beta3 mRNA after orthodontic tooth movement in the two groups were measured and compared.

RESULTSThere were weak positive signals on the cytoplasm of osteoclasts in periodontum in both groups after 12 hours and 3 days force activation. No positive signals were detected in the rest samples. There was no difference in the OD of positive stained osteoclasts between normal and periodontitis groups. Strong expressions were present on cells with one or two nuclei in the alveolar marrow.

CONCLUSIONIt is suggested that integrin beta3 mRNA is related with osteoclasts maturation and migration in orthodontic tooth movement.

Animals ; In Situ Hybridization ; Integrins ; Osteoclasts ; Periodontitis ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Tooth Movement Techniques

OBJECTIVETo initially evaluate the application of artificial vertebra of n-HA/PA66 in anterior reconstruction of lower cervical spine fracture and dislocation.

METHODSIn this study, 84 patients with lower cervical spine fracture and dislocation received anterior cervical discectomy, spinal canal decompression or subtotal corpectomy, spinal canal decompression and reconstruction by n-HA/PA66 composite artificial vertebral body combined with plate instrumentation. Neurological function was followed up by improvement rate of Frankel and situations of the supporting body was observed by X ray and 3D-CT in 3, 12, 24 months postoperatively. The intervertebral height, physical arc (reflected by Cobb angle) and the locations and fusion rate of the supporting body were assessed in order to evaluate the stability of the cervical spine and alignment improvements.

RESULTSAll the patients underwent operation successfully and were followed up for 6 to 24 months with an average of 12 months. The preoperative symptoms were improved to varying degrees. Imaging studies showed that in all cases graft fusion were achieved, and cervical alignments, intervertebral height, cervical spine stability and the locations of the artificial vertebral body were well maintained. No displacement and subsidence of the artificial vertebral body occurred. Postoperative immediate intervertebral height (2.4 ± 0.2) cm, preoperative intervertebral height (1.9 ± 0.1) cm, comparisons of the two groups was statistically significant (q = 2.48, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group intervertebral height was not statistically significant (P > 0.05). Preoperative Cobb angle was 9.8° ± 1.2°, postoperative immediate Cobb angle was 16.6° ± 1.2°, comparisons of the two groups was statistically significant (q = 14.25, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group Cobb angle was not statistically significant (P > 0.05).

CONCLUSIONSn-HA/PA66 artificial vertebral body can provide early cervical spine support and stability and effectively maintain the biological alignment and cervical intervertebral height. It has high rate of graft fusion and is convenient to observe by X-ray. Therefore, n-HA/PA66 can be taken as an ideal graft for anterior lower cervical spine fracture and dislocation operation, but further follow-up study is still required to evaluate the long-term effects.

Adolescent ; Adult ; Aged ; Bone Substitutes ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Hydroxyapatites ; Joint Dislocations ; complications ; surgery ; Male ; Middle Aged ; Nanostructures ; Nylons ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; instrumentation ; Young Adult

OBJECTIVETo evaluate the effect of Taohong Siwu Tang and its fractions on hotplate-induced pain in mice, acetic acid-induced writhing response, dysmenorrheal model and isolated uterine contraction in vitro in mice, and discuss material basis of effect sites.

METHODThe samples of fractions were prepared by macroporous adsorptive resins (TH-1-TH-15). In the whole animal experiment, the hotplate-induced pain mice model was established to observe the effect of the samples on pain threshold in mice; the acetic acid-induced writhing response mice model was built to observe the effect of the samples on the writhing response in mice; the mice dysmenorrheal model was established to observe the effect of the samples on the writhing response, and calcium ion (Ca2+) and nitric oxide (NO) levels in uterine tissue of mice. In the isolated uterus contraction experiment, the oxytocin-induced isolated uterus contraction mice model was established to observe the effect of the samples on the isolated uterus contraction index. HPLC-DAD method was adopted for the content determination of effect sites.

RESULTAccording to the evaluation of the integration geological effect, beside TH-2 and TH-4, other three active fractions (TH-9, TH-10 and TH-11) extracted from Taohong Siwu Tang are the main effect sites. Their chemical components were analyzed and identified as monoterpene glycosides, phthalides, organic acids, etc.

CONCLUSIONThe effect sites of Taohong Siwu Tang on dysmenorrhea are TH-9, TH-10 and TH-11, which are 30% - 50% active fractions obtained from water-soluble small-molecular fractions by gradient elution using ethanol through macroporous absorption resin. Compared with TH-10 and TH-11, TH-9 shows stronger effect, which may be related to the type and content of chemical components it contains.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; therapeutic use ; Dysmenorrhea ; drug therapy ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred ICR

Objective To develop a knowlodge-attitude-practice questionnaire (KAPQ) for abdominal surgery patients,in order to provide an available tool to evaluate the KAP status of patients' postoperative pain.Methods The KAPQ was established on the basis of clinical experience,literature review and expert consultation.Ninety patients with abdominal surgery in a three level hospital in Jiangsu province were chosen by convenient sampling and SPSS was used for the scale reliability and validity.Results The internal consistency reliabilities of the KAPQ and its three subscales were respectively 0.899,0.933,0.903 and 0.888.The copy of reliability of three subscales were respectively 0.886,0.763 and 0.821.The content coefficients of three subscales were respectively 0.946,0.921 and 0.949.Factor analysis showed that the questionnaire had good construct validity with a common explaining variance more than 0.4.Conclusions The KAPQ has good reliability and validity,thus it can be used as an effective tool to evaluate the health-related knowledge-attitudepractice of the abdominal surgery patients' postoperative pain.

Objective To detect human parechovirus (HPeV) from stool samples of hospitalized children for acute gastroonteritis of undetectable etiology. Methods We conducted a real-time PCR to detect HPeV. Result The results showed that 24 of 99 (24%) children with gastroenteritis of undetectable etiology were detected with HPeV. Four known HPeV types (HPeV1,3,4,6) were detected in the present study. HPeV1 (50%) was frequently identified as the predominant strain and follow by HPeV3 (25 % ), HPeV 4 (8.3 % ) and HPeV 6 (4.2 % ). We were unable to type 3 samples. Conclusion HPeV was prevalent in hospitalized children for acute gastreenteritis of undetectable etiology in China. Further study is needed for clarifying the role of HPeV in gastroenteritis.