It has been found that the hypofunction status of hypothalamus-pituitary-adrenal (HPA) axis exists in patients with Shen-yang deficiency syndrome of TCM, also presents in most asthma patients. Seasonal attack of asthma can be prevented with Shen-tonifying drugs by improving adrenocortical function. Since patients subject to long-term glucocorticoids display hypofunction condition of HPA axis, Shen-tonifying drugs should be helpful to gluocorticoid withdrawal for getting higher success rate. Basic researches also indicated that the activating of adrenocortical stem cells and promoting regeneration of adrenal cortex is one of the mechanisms underlying improvement of adrenocortical function. Series of research showed that hypofunction of adrenocortex is the general pathological change in some diseases, so, Shen-tonifying drugs act a part in unitarily modulating the adrenocortical function, to get the therapeutic effect of both regulating the whole and improving the local.
Adolescent ; Adult ; Asthma ; physiopathology ; therapy ; Child ; Female ; Humans ; Hypothalamo-Hypophyseal System ; physiology ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Nephrotic Syndrome ; physiopathology ; therapy ; Pituitary-Adrenal System ; physiology ; Yang Deficiency ; physiopathology ; Young Adult

This study was purposed to construct three siRNA eukaryotic expression vector specific to mouse Qa-1 gene, to investigate its silencing effect on Qa-1 gene and to select the most efficient siRNA plasmid specific to mouse Qa-1 gene. Three siRNA peptides specific to mouse Qa-1 through siRNA Web design tools of Ambion company were chosed. Jingsai Company helped to complete the siRNA eukaryotic expression vector. The mouse NIH3T3 cells cultured in RPMI 1640 medium with 10% fetal bovine serum were divided into four groups: three groups of the cells were transfected with lipofectamine 2000 reagent and three different siRNA eukaryotic expression vectors, while one group cells were transfected with lipofectamine 2000 reagent alone as negative control. Cells were collected at 24, 48, 72 hours after transfection; the RNA level of Qa-1 was detected by RT-PCR, and the expression position was examined with flow cytometry analysis by using anti-Qa-1 monoclonal antibody. The results indicated that the constructed three siRNA eukaryotic expression vectors were found to be specific to mouse Qa-1 gene. The sequence analysis showed that the sequence was identical to what chosed from web tools. NIH3T3 cells in vitro were adhered in culture that cell shape appeared to change after transfection. RT-PCR and flow cytometry analysis by using anti-Qa-1 monoclonal antibody approved that both Qa-1 RNA and the expression of Qa-1 on cell surface decreased. The decreased levels in the three groups were different. At 24, 48 and 72 hours, the expression of Qa-1 on NIH3T3 cells decreased as in the following: H2-T231: 60.9%, 81.9%, 43.6%; H2-T232: 64.5%, 73.9%, 61.1%; H2-T233: 61.9%, 71.2%, 47.5%. H2-T232 was most efficient one in all three time points. It is concluded that all three siRNA eukaryotic expression vectors selected can successfully suppress the expression of the Qa-1, and from them H2-T232 is most efficient.
Animals ; Base Sequence ; Cells, Cultured ; Gene Silencing ; Genetic Vectors ; Histocompatibility Antigens Class I ; genetics ; Mice ; NIH 3T3 Cells ; Plasmids ; RNA, Small Interfering ; genetics ; Transfection

Objective To explore the therapeutic efficacy of levofloxacin combined with anti-tuberculosis drugs and thoracic catheterization for the treatment of tuberculous pleuritis.Methods Patients who were admitted to Departments of Infectious Diseases of Hanzhong Central Hospital and Ankang Central Hospital between February 2014 and August 2016 for initial treatment of tuberculous pleuritis were included in the study,they were divided into groups A,B,C and D.Group A received 2HRZE + 7HR regimen combined with conventional drainage;group B received 2HRZE+ 7HR regimen combined with thoracic catheterization;group C received 2HRZEV + 7HR regimen combined with thoracic catheterization;group D received 2HRZEV + 10HR regimen combined with thoracic catheterization.groups B,C and D received thoracic catheterization,normal saline 20mL and urokinase 100,000U were given through the drainage tube.Results A total of 172 patients with newly diagnosed tuberctlous pleurisy were received for treatment.There were 45,53,38,and 36 cases in group A,B,C,and D respectively.The total effective rate of therapy for pleural effusion in group A was lower than that in group B(64.44％ vs 90.57％,x2 =9.863,P＜ 0.05);after two month therapy,total effective rate of therapy for pleural effusion in group B was lower than that in group C (18.87％ vs 39.47％,x2 =4.716,P＜0.05);at the end of therapy,total effective rate in group C was lower than that in group D (60.53 ％ vs 83.33 ％,x2 =4.731,P＜0.05).Conclusion For initial treatment of patients with tuberculous pleuritis,2HRZEV + 10HR antituberculosis regimen combined with thoracic catheterization and urokinase infusion can significantly improve the clinical symptoms and recovery rate of tuberculous pleuritis,facilitate drainage of pleural effusion and prevent pleural thickening,adhesion and encapsulation.

BACKGROUND:High ankle sprain is easily missed and leads to ankle dysfunction.Arthroscopy can detect hidden high ankle sprain.Suture-button elastic fixation can restore the biomechanical stability of the distal tibiofibular syndesmosis. OBJECTIVE:To explore the clinical efficacy of Suture-button elastic fixation for high ankle sprain under ankle arthroscopy. METHODS:A retrospective analysis was performed on 40 cases of high ankle sprain patients treated with Suture-button elastic fixation under ankle arthroscopy from August 2019 to August 2021 in the Department of Foot and Ankle Surgery,Wuhan Fourth Hospital.All patients underwent Suture-button elastic fixation.The American Orthopedic Foot and Ankle Society function score,Visual Analog Scale pain score,ankle range of motion,preoperative imaging data,and arthroscopic tibiofibular syndesmosis separation degree were recorded.Meislin criteria were used to evaluate the curative effect and postoperative complications were recorded. RESULTS AND CONCLUSION:(1)40 patients were followed up for 16-48 months after operation.(2)At the last follow-up,American Orthopedic Foot and Ankle Society score was(88.95±6.64 points).Visual Analog Scale score was(1.78±1.23 points).Ankle dorsiflexion range of motion was(33.50±5.79 degrees).Ankle plantarflexion range of motion was(34.50±5.97 degrees).There were statistically significant differences before and after surgery(P<0.05).(3)There was a low positive correlation between the radiographic separation index and the degree of arthroscopic separation(r=0.612,P<0.01).(4)The curative effect was evaluated by Meislin standard,with an excellent and good rate of 95%(38/40).Postoperative ankle joint pain was relieved,and ankle joint activities were significantly improved.(5)During the follow-up period,all patients had no nerve injury or incision infection.In 1 patient,the internal fixation was removed due to skin irritation and squatting sensation after operation.(6)It is concluded that Suture-button elastic fixation for high ankle sprain is effective under ankle arthroscopy in restoring ankle function and maintaining joint stability without the need for secondary removal,and it is worth clinical application.

Objective:To further develop Gaultheria leucocarpa var. yunnanensis,anti-bacteria constituents in it were　screeened. Method:The constituents were extracted by chromatographic process.The anti-bacteria test was made with regulatory method of analysis. Result and conclusion: Anti-bacteria test with extracts of water, acetic ester and n-butanol showed that 3 extracts from 22 samples had anti-Staphylococcus aureus action, and the extracts from root and stem showed the same result. 2 extracts 　could kill Escherichia coli and Pseudomonas aeruginosa. The lower the concentrate, the less the anti-bacteria action was. These results suggested that not only essential oil but other ingredients from 　G.leucocarpa var. 　yunnanensis have anti-bacteria activity . Anti-fungi test of the same extracts didn't indicate remarkable action.

BACKGROUNDThe aim of this study was to investigate the potential relationship between the dynamic expression of Toll-like receptor 2 and 4 (TLR2/4) in peripheral blood mononuclear cells as well as changes in serum concentration of inflammatory factors and acute lung injury (ALI) in patients after orthotopic liver transplantation (OLT).

METHODSThe peripheral blood samples of 27 patients (23 men and 4 women with ASA III to IV) who received OLT were collected for measurement of TLR2/4 at T1 (after induction of anesthesia), T2 (25 minutes after anhepatic phase), T3 (3 hours after graft reperfusion) and T4 (24 hours after graft reperfusion). The expression of TLR2/4 in mononuclear cells was measured by flow cytometry. The serum concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-8 were measured by enzyme-linked immunosorbent assay (ELISA). Twenty-seven patients were assigned to ALI group (n = 9) and non-ALI group (n = 18) according to the diagnostic criteria of ALI. The expression of TLR2/4 in the ALI group or non-ALI group was analyzed.

RESULTSCompared to the non-ALI group, the volumes of blood loss, ascites, total output and transfused red blood cells were higher in the ALI group, and the anhepatic phase lasted longer (P < 0.05, P < 0.01). The expression of TLR2/4 in mononuclear cells increased significantly at T3 and T4, and serum concentrations of TNF-alpha, IL-1beta and IL-8 increased significantly too. There was no significant difference in Child-Turcotte-Pugh (CTP) scores between the ALI group and non-ALI group (P > 0.05). The expression of TLR2/4 in mononuclear cells increased significantly at T3 and T4 in the ALI group (P < 0.05, P < 0.01). A positive correlation was noted between the expression of TLR4 in mononuclear cells and the serum concentrations of TNF-alpha, IL-1beta (P = 0.041, P = 0.046) in the ALI group. In the non-ALI group, statistical results showed that the expression level of TLR2/4 in mononuclear cells was not significantly different during the peri-operative period of OLT (besides TLR4 expression at T4). Compared to the non-ALI group, the increasing amplitude of TLR2/4 expression in mononuclear cells was more significant in the ALI group. The patients whose TLR2/4 expression in mononuclear cells exceeded that at T1 by one time were more likely to suffer from ALI (P = 0.013), with a relative risk of 16.

CONCLUSIONThe expression level of TLR2/4 in mononuclear cells increases significantly in the peri-operative period of OLT, and it may be a high risk factor for occurrence of postoperative ALI.

Acute Lung Injury ; etiology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Liver Transplantation ; adverse effects ; Male ; Postoperative Period ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

This study was aimed to investigate the biological characteristics of osteoblasts and their hematopoietic supportive function by using human fetal osteoblastic cell line 1.19 (hFOBs) as a model. The pluripotency markers (Oct-4, Rex-1, hTERT) of hFOBs were analyzed by RT-PCR, the multilineage differentiation experiments were conducted in vitro. Flow cytometry (FCM) was used to identify the surface markers of hFOBs, and RT-PCR was used to analyze their hematopoietic cytokine expression in comparison with bone marrow mesenchymal stem cell (BM-MSC). The results showed that hFOBs expressed several ESC pluripotency markers including Oct-4 and Rex-1, except hTERT. Moreover, hFOBs could also undergo multilineage differentiation into the mesodermal lineages of adipocytic cell types in addition to its predetermined pathway, the mature osteoblast. Both hFOBs and BM-MSC expressed CD44, CD73 (SH3), CD105 (SH2) and CD90 (Thy1), and lack expression of CD34, CD45, or HLA-DR surface molecules. In addition, both hFOBs and BM-MSC expressed SCF, IL-6, and SDF-1alpha mRNA, but only hFOBs could express GM-CSF and G-CSF. It is concluded that human fetal osteoblastic cell line 1.19 may provide a good model to study the osteoblastic regulation role in hematopoiesis in vitro.
Cell Differentiation ; physiology ; Cell Line ; Fetus ; Hematopoiesis ; physiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology ; Models, Biological ; Osteoblasts ; cytology ; physiology

BACKGROUNDLive attenuated hepatitis A vaccine (H2 strain) is widely applied in prevention of hepatitis A epidemic in China and other countries now. It is essential to observe and confirm the vaccine immune efficacy, population antibody level and its persistent efficacy after mass immunization.

METHODSA total of 220 children with negative anti-HAV antibody (aged 1 - 3 years) were taken for follow-up assay to observe seroconversion and geometric mean titre (GMT) level 2 months, 12 months, 6 years, and 10 years after inoculation. Another survey sampled from subjects of different age groups (3, 6, 9, 15, 18, 25 and 35 years) to compare anti-HA antibody positive rate before and after inoculation performed 10 years previously. Epidemiological observations were taken for 10 years to evaluate the relationship between vaccine coverage and hepatitis A morbidity. Serum antibody to HAV was detected by enzyme linked immunoassay (ELISA, calibrated by WHO international reference) and ABBOTT Axsym HAVAB microparticle enzyme immunoassay.

RESULTSSeroconversion in follow-up assay 2 months and 10 years after inoculation was 98.6% and 80.2% respectively. For children, the vaccination anti-HA antibody positive rates were significantly different before and after 10 years, 7.69% cf 70.45% (aged 3 years) and 52.58% cf 71.78% (aged 18 years). When vaccine coverage rose from 57% to 74%, there were no any HA epidemics. When vaccine coverage reached 85%, there were no any HA cases. With vaccine coverage between 85% and 91%, there were no any HA cases in cohorts from the age of 1 year to 15 years during the 10 years.

CONCLUSIONSLive attenuated hepatitis A vaccine has an obvious long-term effectiveness in prevention and control of HA epidemics through mass vaccination.

Adolescent ; Adult ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Follow-Up Studies ; Hepatitis A ; prevention & control ; Hepatitis A Vaccines ; immunology ; Hepatitis Antibodies ; blood ; Humans ; Immunoglobulin G ; blood ; Mass Vaccination ; Vaccines, Attenuated ; immunology

OBJECTIVETo control the quality of Gaultheria leucocarpa var. yunnanensis the contents of lignans in these plants from different provinces were detected.

METHODHPLC was adopted using Hypersil-18 column (0.4 cm x 25 cm), methanol: acetonitrile: water (30:5:65) as mobile phase.

RESULTThe calibration curves of D1(gaultherinA), D2[(-)-5'-methoxyisolariciresinol-2 alpha-O-beta-D-xylopyranoside)], D4[(+)-lyoniresinol-2 alpha-O-beta-D-glucopyranoside)] were linear in the range of 0.10-0.40 microgram, 0.03-0.13 microgram, 0.02-0.09 microgram, respectively. The average recovery of D1 was 98.9% and RSD was 2.6%.

CONCLUSIONComparing the samples from Jinxiou(158.2 mg.100 g)-1 and Kumming(154.8 mg.100 g)-1, that of Guiyang showed the highest content of LD(208.8 mg.100 g)-1. The content of LD in the tender above-ground part was 1.3 times higher than that in the wilt.

Benzoates ; analysis ; Chromatography, High Pressure Liquid ; Gaultheria ; chemistry ; Glycosides ; analysis ; Lignans ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control

OBJECTIVEHemolytic uremic syndrome (HUS) is a common primary disease that can cause acute renal failure in childhood. Renal disease is the most important long-term complication in patients who survived the acute stage of HUS. Use of angiotensin-converting enzyme inhibitors (ACEI) and a restricted protein intake may be beneficial to the patients. However, it is not established whether such patients should be treated with steroids and immunosuppressors. The present study aimed to probe into the benefit of using steroid and immunosuppressor in patients after acute stage of HUS.

METHODSThe subjects included 17 patients (aged 9 months to 15 years, 12 males, 5 females) with HUS. Thirteen patients recovered from the acute stage of HUS, and underwent continuative treatment and follow-up. All the patients were treated with ACEI and early restriction of protein intake. Additionally, 2 children manifested as glomerulonephritis, one was treated with triperygium glycosides. Other 11 children who manifested as nephrotic syndrome were treated with prednisone, among them 5 children had no response or had incomplete response to prednisone, for these children short-term high dose cyclophosphamide or methylprednisolone pulse treatment were added; in 3 of the children short-term high dose methylprednisolone treatment was applied additionally for membranoproliferative glomerulonephritis and/or focal segmental glomerulosclerosis and crescentic glomerulonephritis.

RESULTSAfter follow-up for 2 months to 8 years, 4 patients with milder disease recovered, their blood pressure, renal function and urinalysis became normal, but 1 patient had recurrence. Among 9 patients with severe disease, 6 maintained normal blood pressure, recovered renal function and urinalysis, the other 3 patients failed to comply with treatment protocol and died during the 3rd, 9th and 13th month. The remainder (4 cases) gave up therapy and died on the 27th to 48th days of the course.

CONCLUSIONThe treatment applied in this study could improve the prognosis of patients after acute phase of HUS evidently by using the steroid and immuno suppressor according to clinical classification and pathological findings. It is recommended that triperygium glycosides is beneficial to children with glomerulonephritis, proteinuria and hematuria after acute stage of HUS. Adjustment of therapeutic schedule based on pathological findings after renal biopsy is helpful. To the patients with progressive renal failure who have no response to the steroid and immunosuppressors, steroid and immunosuppressor should be discontinued and dialysis treatment should be applied. Protocol compliance is also an important factor.

Acute Disease ; Adolescent ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Child ; Child, Preschool ; Combined Modality Therapy ; Diet, Protein-Restricted ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Hemolytic-Uremic Syndrome ; diet therapy ; drug therapy ; physiopathology ; Humans ; Immunosuppressive Agents ; therapeutic use ; Infant ; Male ; Prognosis ; Steroids ; therapeutic use ; Treatment Outcome