ObjectiveTo establish a method to induce heterotopic ossification (HO) after spinal cord injury. Methods10 New Zealand White rabbits underwent a complete transection of the thoracic (T10) spinal cord. Just after transection, the right hind limb of every rabbit was immobilized with a plaster support to keep the knee in extension and the hip unrestricted. The plaster was temporarily removed once a day (six times per week) to allow the knees passively mobilized at the maximum range for 5 min. Local swelling, local skin temperature and grade of ossification of the paraplegic limbs were observed. After 5 weeks, the rabbits were sacrificed, and the quadriceps muscles of the right hind limbs were removed for the histological examination. ResultsAfter 5-week immobilization and temporary passive mobilization, HO was successfully induced in all 10 rabbits. There was local skin temperature increase and local swelling in the right hind limbs. The histological changes were similar to those observed in clinical HO. Conclusion5-week immobilization and temporary passive mobilization approach may successfully induce HO, and it may be used to study the pathogenesis of HO after spinal cord injury.

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

OBJECTIVE: To compare the hemostatic effects of local packing of Nasopore combined with hemocoagulase injection and local packing of Nasopore combined with saline injection for postoperative management of functional endoscopic sinus surgery by a double-blind, randomized control clinical trial. METHOD: Sixty-eight cases of chronic sinusitis needed functional endoscopic sinus surgery were randomly divided into the experimental group of 40 cases and control group of 28 cases, respectively. For the experimental group, 1 U of hemocoagulase dissolved in 0.5 ml saline was injected into Nasopore which was packed into the nasal cavity after operation. For the control group, 0.5 ml of saline was injected. The postoperative bleeding of the two groups were scored by visual analogue scale. RESULT: There was statistically significant difference between the bleeding VAS scores assessed 6 hours and the ones assessed 1, 2 and 3 days after the operation in the control group (P < 0.05). There was the statistically significant difference between the bleeding VAS scores of experimental group and control group assessed 6 h after the operation (P < 0.05). CONCLUSION The hemocoagulase may improve the hemostatic effect of Nasopore 6 hours after the operation by combined injection with Nasopore as nasal cavity packing.
Bandages ; Batroxobin ; administration & dosage ; Double-Blind Method ; Endoscopy ; Epistaxis ; therapy ; Female ; Humans ; Injections ; Male ; Nasal Cavity ; surgery ; Treatment Outcome

OBJECTIVETo study the effect and mechanism of Ganbi decoction (GBD) in treating patients with antituberculotic agent caused liver injury (ATB-LI).

METHODSOne hundred and twenty-eight patients with ATB-LI were randomly assigned to the treated group (n = 66) and the control group (n = 62) with the envelop method. Meanwhile, 60 healthy persons were selected as the healthy control group. The treated group was treated by GBD one dose every day with the constituents modified depending on patients' symptoms, and the control group was treated with glucuronolactone tablets and inosine injection. One week was taken as one treatment course. The changes of clinical syndromes, physical signs, T-lymphycyte sub-groups and serum level of nitric oxide (NO) were observed before and after treatment and the recovery time of liver function was recorded. The outcome was compared with that in the healthy control group.

RESULTSIn the treated group, 28 patients (42.4%) were cured, 30 (45.5%) improved and 8 (12.1%) ineffectively cured, the total effective rate being 87.9% (58/66). In the control group, 17 patients (27.4%) were cured, 24 (38.7%) improved, and 21 (33.9%) ineffectively cured, the total effective rate being 66.1% (41/62). The total effective rate in the treated group was significantly higher than that in the control group (P < 0.05). Liver function was improved in both groups, recovery time in the treated group was 12.0 +/- 7.0 days, which was significantly shorter than that in the control group (16.0 +/- 8.0 days), showing significant difference between the two groups (P < 0.05). The levels of CD3, CD4 and CD8 were significantly higher and level of NO significantly lower in the two groups of patients than those in the healthy control group (P < 0.05), but these parameters were improved more significantly in the treated group after treatment, when compared with those before treatment or with those in the control group, all showing significant difference (P < 0.05).

CONCLUSIONGBD could prevent ATB-LI, and its mechanism could be by way of reducing NO production induced by endotoxin of macrophage and stimulating the proliferation of T-lymphycyte to elevate immunity.

Adult ; Aged ; Antitubercular Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucuronates ; therapeutic use ; Humans ; Inosine ; therapeutic use ; Liver Diseases ; drug therapy ; Liver Function Tests ; Male ; Middle Aged ; Nitric Oxide ; blood ; T-Lymphocyte Subsets ; Treatment Outcome

OBJECTIVETo study the anticancer effect in vitro and in vivo and mechanism of DHA compound.

METHODSCervical cancer cell line HeLa cells, glioma cell line U251 cells and mouse hepatoma H(22) tumor were used in this study. Transmission electron microscopy and fluorescence microscopy were used to observe the morphological changes of cell apoptosis. Western blot was used to detect the expression of caspase-3. RT-PCR was used to determine the effect on Bcl-2 and Bax mRNA transcription in U251.

RESULTSAntitumor effect was observed in vivo and in vitro. Typical morphological changes were seen in cancer cells. The level of caspase-3 was significantly increased and the content of Bcl-2 mRNA was decreased significantly, while the content of Bax mRNA was significantly increased in the U251 cells after treatment with DHA compound.

CONCLUSIONDHA compound can inhibit the growth of some types of tumors and the increase of caspase-3 and Bax mRNA and decrease of Bcl-2 mRNA may be involved in its mechanism of action.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Glioma ; pathology ; HeLa Cells ; Humans ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

This paper reviewed the progress in researches on antiviral activity of chemical constituents from plants in recent years, the antiviral activity and mechanism of action of flavonoids, alkaloids, terpenoids, coumarins and polysaccharoses were sammarszed, provided new leading compound for antivirus new drugs from the plares in prospect.
Antiviral Agents ; chemistry ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Molecular Structure ; Plant Extracts ; chemistry ; pharmacology

OBJECTIVETo investigate the anti-tumor effect of dendritic cells (DC) pulsed with G422 glioblastomas RNA in mice bearing intracranially G422 glioblastomas.

METHODSDCs were pulsed in vitro with glioblastomas G422 cell RNA. The tumor-bearing mice were injected intratumorally or subcutaneously with pulsed DCs, PBS, non-pulsed DCs. The survival duration of mice was recorded. Serum levels of cytokine IFN-gamma, IL-2, IL-10, IL-4 were detected. Pathological examination was performed.

RESULTSThe survival duration of mice with DC-based vaccine increased significantly(P<0.01). The serum IFN-gamma level was increased (P<0.01) and IL-10 level was decreased (P<0.05) after treatment. Pathological examination showed necrotic tumor in the treatment mice.

CONCLUSIONDC vaccination can significantly increase survival duration of mice with intratumoral or subcutaneous administration of vaccines.

Animals ; Brain Neoplasms ; immunology ; therapy ; Cancer Vaccines ; immunology ; Dendritic Cells ; immunology ; transplantation ; Glioblastoma ; immunology ; therapy ; Immunotherapy ; methods ; Mice ; RNA, Neoplasm ; immunology ; Random Allocation ; T-Lymphocytes, Cytotoxic ; immunology

The applicator therapy is a unique method to treat infant diarrhea in traditional Chinese medicines and widely applied in clinical practice. Currently, many researchers have proved the rationality of the therapy based on the traditional Chinese medicine mechanism and on the data from clinical practice, but its action mechanism is uncertain at present. In this study, with the assistance of pediatric practitioners, the automated ribosomal intergenic-spacer analysis (ARISA) was adopted to study the effect of the adjuvant therapy with Dingguier umbilical paste on intestinal flora of diarrhea infants, in which Dingguier umbilical paste served as the adjuvant therapy in oral traditional Chinese medicines and fecal samples of infants with different diarrhea symptoms were collected and used as the study materials. The results showed that the adjuvant therapy had a significant effect on the shift of intestinal flora, which was associated with the decrease in the similarity difference to the normal control group and the increase in the number of operational taxonomic units (OTUs) shared with the normal control group. Additionally, adjuvant therapy with Dingguier umbilical paste also showed long action duration and increased OTUs number. These results indicated that Dingguier umbilical paste has the effect in restoring the micro-ecosystem of unbalanced intestinal bacteria. Intestinal flora may be one of major targets for the applicator therapy for the infant diarrhea, but not for the single oral traditional Chinese medicine for infant diarrhea.
Adjuvants, Pharmaceutic ; therapeutic use ; Diarrhea ; drug therapy ; microbiology ; Feces ; microbiology ; Female ; Humans ; Infant ; Intestines ; drug effects ; microbiology ; Male ; Medicine, Chinese Traditional ; methods ; Ointments ; Treatment Outcome ; Umbilicus

ObjectiveTo compare the efficacy of intravenous infusion of dexmedetomidine and midazolam for premedication in children.MethodsNinety-two ASA Ⅰ or Ⅱ children (46 cases aged 1-3 yr and 46 cases aged 4-6 yr) scheduled for elective general or urologic surgeries,were enrolled in this study.The children were randomly divided into 2 groups (n =46 each):midazolam group (group M) and dexmedetomidine group (group D).The children accompanied by their parents were admitted to the anesthesia preparation room at about 20 min before induction of anesthesia,and midazolam 0.1 mg/kg òr dexmedetomidine 1 μg/kg was infused intravenously over 10 min.Anesthesia was induced with proporol-rocuroniume-remifentanil,and maintained with sevoflurane-remifentanil-rocuroniume.Modified Yale Preoperative Anxiety Scale (mYPAS) score,sedation score,HR,mean arterial pressure (MAP),respiratory rate (RR) and SpO2 were recorded before premedication (T1),before separation from their parents (T2) and when entering the operating room (T3).The incidence of sleep (a sedation score of 4) was recorded at T2,3.The end-tidal concentration of sevoflurane,infusion rate of remifentanil,laryngeal air way removal time,emergence time,duration of stay at the recovery room,incidence of delirium during recovery period,the percentage of patients requiring rescue analgesic,and adverse effects were also retorded.ResultsCompared to that at T1,the mYPAS score was significantly decreased at T2,3,and the sedation score was significantly increased at T2,3 in both groups ( P ＜ 0.05),HR at T2 and MAP at T2,3 were significantly decreased in group D,and HR at T3 was significantly increased in group M ( P ＜ 0.05 ).Compared to group M,the sedation scores and the incidence of sleep were significantly increased at T2,3,and the HR was significantly decreased at T2 in group D ( P ＜ 0.05).There was no significant difference in the mYPAS score,RR,MAP,SpO2,end-tidal concentration of sevoflurane,infusion rate of remifentanil,laryngeal air way removal time,emergence time,duration of stay at the recovery room,incidence of delirium during recovery period,the percentage of patients requiting rescue analgesic,and incidence of adverse effects between D and M groups ( P ＞ 0.05).ConclusionThe sedative efficacy of iv dexmedetomidine is superior to that of iv midazolam when infused for premedication in children,but it exerts much influence on hemodynamics,and the changes in hemodynamics should be noted.

For diabetes mellitus, little research has been done on the tissue-based or cell-based drug screening model, which has advantages over traditional animal diabetic model in high specificity, high screening volume, low cost and simple manipulation. Considering that the maintenance of complete islet tissue structure is the prerequisite for islet cells to perform their functions normally, an in vitro islet-based drug screening model for diabetes mellitus was established and evaluated. Pancreatic islets were isolated from 3 weeks old mice of either sex by collagenase digestion and density gradient centrifugation as prescribed by Ramanadham S. The volume of 0.1% (W/V) collagenase IV, 0.1% (W/V) Hyaluroridase and 0.1% (W/V) DNase I were 4 times, 2 times and 1 times that of the islets to be digested. And a 2 hours' cold digestion at 4 degrees C was followed by a 10 minutes' warm digestion at 37 degrees C. Under the optimized digestion condition, the islet recovery could be increased by 10%. The isolated islets could survive 6 weeks in vitro and show stable insulin secretion in the first 10 days after inoculation. The obtained islets were cultured in RPMI-1640 medium at 37 degrees C with 5% CO2. Then a diabetic model was established by selecting streptozotocin (STZ) as the evocator and nitric oxide (NO) as the responding index. After 1 day's inoculation, islets culture was treated with STZ, whose concentration ranged from 0 to 5.0 mmol/L. NO was measured by a colorimetric assay at 540nm based on the Griess reaction for 10 min with 0.1 mL Griess reagent and 0.1 mL culture supernatants. Insulin secretion was assayed by RIA methods. Due to the islets-related inoculation variations, NO release and insulin content were both expressed as a percentage of the value recorded in basal experiment which was in the only presence of Krebs culture medium. It was testified that the amount of NO released from islet itself remained steady at 30-35 mmol/L regardless of the changes of STZ concentration from 0 to 5.0 mmol/L. However the NO content in the supernatants of islets culture had close relationship with STZ concentration. This indicated that in this STZ-induced islet diabetic model, NO mainly comes from STZ when it dissolves in water. On the other hand, when STZ changed from 0 to 5.0 mmol/L, the dose-dependent relationship between NO content and insulin secretion showed that the increase of NO came along with the decrease of insulin secretion, which is an important symbol of islet function. As a kind of oxidative free radical, NO is capable of impair islet cells. Thus, NO is a reliable responding index of the model. The optimal STZ concentration in the model is finally determined to be 5.0 mmol/L, under which condition the NO content and insulin secretion is 10.81 times and 0.43 times that in the medium before STZ is added. So if anything is effective in lowering the NO content in the culture, it could protect islets cells from the oxidative attacks of NO. Finally, as an application of the model, the scavenging effect of KOSCr on NO was studied. In a series of KOSCr with different chromium content, all had shown better NO scavenging effects than KOS itself, which could give us an enlightenment of the influence of chromium ion on oligosaccharide. And 1 g/mL KOSCr with 3.519% chromium content can significantly inhibit the NO formation. This has lain a theoretic basis for the research of KOSCr bioactivity and quality control. These results suggested that the STZ-induced diabetic islet model which is impaired by NO free radical can be used effectively, fast and conveniently when screening potential diabetes drugs.
Animals ; Chromium ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; Disease Models, Animal ; Female ; In Vitro Techniques ; Insulin ; metabolism ; Islets of Langerhans ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Nitric Oxide ; metabolism ; Streptozocin ; pharmacology