0.05), but the level of albumin dropped significantly(P

Abstract: Objective To explore and analyze the diagnostic value of multicolor melting curve analysis (MMCA) for the resistance of five anti-tuberculosis drugs, so as to clarify the clinical value of MMCA in detecting drug resistance of Mycobacterium tuberculosis. Methods From April 2021 to May 2022, 200 patients with positive Mycobacterium tuberculosis admitted to the Fourth People's Hospital of Qinghai Province were selected as research objects, and sputum specimens were taken from the patients. Traditional Mycobacterium tuberculosis drug sensitivity test (modified Löwenstein-Jensen medium method) and MMCA analysis were respectively given to detect the resistance of five anti-tuberculosis drugs, including isoniazid, ethambutol, streptomycin, rifampicin and isoniazid, respectively. Those samples with inconsistent results between the two diagnosis methods were subjected to gene sequencing verification, and the diagnosis efficiency of MMCA for the five anti-tuberculosis drugs was compared. Results Using Mycobacterium tuberculosis drug sensitivity as the gold standard for drug resistance diagnosis, the sensitivity of MMCA for detecting drug resistance of rifampicin, ethambutol, streptomycin, isoniazid and levofloxacin were 95.83% (46/48), 93.75% (15/16), 100.00% (15/15), 100.00% (20/20) and 70.00% (7/10), respectively, with statistical differences between groups (P<0.05). There were no statistically significant differences in the specificity, positive predictive value, negative predictive value and accuracy of MMCA for the five anti-tuberculosis drugs (P>0.05). For the 8 samples with inconsistent results between MMCA and modified Löwenstein-Jensen medium method, gene sequencing was performed and compared with the results of gene sequencing. After comparison with gene sequencing results, it was found that the coincidence rate of MMCA and gene sequencing results was 75.00% (6/8). Conclusions In the detection of drug-resistant mutations in TB patients, multi-color probe fusion curve analysis has high diagnostic efficacy for first-line anti-tuberculosis drugs, but is not sensitive to second-line anti-tuberculosis drug levofloxacin. Therefore, for the detection of first-line anti-tuberculosis drugs, MMCA has a good clinical application prospect.

Objective To learn the status of iodine nutrition and the goiter of children and pregnant woman in Zhejiang Province after the iodine salt concentration adjustment.Methods The proportionate to population size sampling was applied to select 30 surveillance areas in Zhejiang Province,and iodine nutritional status were evaluated by 4 indicators that include goiter rate in 8 -10 years old children,urine iodine,iodized salt and water iodine.Results The iodine content of 1 565 edible salt samples collected from the above children was measured,and the iodine median(P25 -P75 )was 23.30 (21.00 -25.80)mg/kg.The coverage of iodized salt rate was 88.43%,and 95.88% iodized salts was qualified.The qualified iodized -salt ingesting rate was 84.79%.The iodine content of children aged 8 -10 years old and pregnant woman was tested,and iodine median (P25 -P75 )was 171.99 (113.00 -243.89 ),134.14 (82.10 -202.98)g/L respectively.The iodine content of 66 water samples were randomly collected from the surveillance areas to be tested,and iodine median(P25 -P75 )was 2.42(1.17 -6.28)μg/L.The proportion less than 10 g/L was 87.9%.The goiter status of 1 565 children were examined by B -type of ultrasonic,and the prevalence of goiter was 4.98%.Conclusion After the iodine salt concentration adjustment,the status of iodine nutrition in children was in optimum level,but the level of iodine nutrition in pregnant women was less than the state,which should be paid more attention.

OBJECTIVETo observe the effect of Wenyang Huazhuo Tongluo Recipe (WYHZTLR) on the proportion of T helper 17 cells (Thl7)/regulatory T cells (Treg), and serum levels of IL-17 and IL-10 in peripheral blood of systemic sclerosis (SSc) patients with yang qi insufficiency and turbidity induced collaterals blockage syndrome (YQITICBS).

METHODSTotally 82 SSc patients were randomly assigned to the Western medicine group (as the control group) and the integrated Chinese and Western medicine group (as the treatment group), 41 cases in each group. All patients took methotrexate (MTX) tablet and prednisone tablet. Patients in the treatment group additionally took WYHZTLR. The treatment course for all was six consecutive months. Besides, another 70 healthy volunteers were recruited as a healthy control group (as the healthy group). Percentages of Th17 and Treg in peripheral blood were detected by flow cytometry. Serum levels of IL-17, IL-10, von Willebrand factor (vWF), aminoterminal propeptide of type l procollagen (PIIINP), and cross-linked carboxyterminal telopeptide of type I collagen ( I CTP) were measured by double antibody sandwich ELISA. The correlations between Th17/Treg and levels of vWF, PIIINP, I CTP, skin score, and disease activity index were observed by Pearson correlation analysis.

RESULTSThe percentage of Th17 in peripheral blood, ratios of Th17/Treg, and the serum level of IL-17 were significantly higher, but the percentage of Treg and the serum level of IL-10 were significantly lower in SSc patients, when compared with those of the healthy group (P <0. 05, P <0. 01). Compared with the same group before treatment, the percentage of Thl7, ratios of Thl7/Treg, and levels of IL-17, vWF, and PIIINP all decreased in the two groups after treatment (P <0.05, P <0.01), but the percentage of Treg and the IL-10 level increased (P <0. 05, P <0. 01). Meanwhile,the level of I CTP was higher in the treatment group after treatment (P <0. 05). The improvement of all indices except the percentage of Th17 was more obvious in the treatment group than in the control group (P <0. 05, P <0. 01). The ratio of Th17/Treg was positively correlated with levels of vWF, PIIINP, skin score, and disease activity index before and after treatment respectively (P <0. 01), but with no obvious correlation with the level of I CTP (P >0. 05).

CONCLUSIONWYHZTLR could achieve its therapeutic effect on SSc patients by regulating Th17/Treg imbalance, lowering levels of vWF and PIIINP, and elevating the level of I CTP.

Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-10 ; Interleukin-17 ; Methotrexate ; Peptide Fragments ; Procollagen ; Scleroderma, Systemic ; drug therapy ; T-Lymphocytes, Regulatory ; Th17 Cells

OBJECTIVETo investigate a specific and sensitive assay for the diagnosis of autoimmune thrombocytopenic purpura (AITP).

METHODSGlycoprotein specific autoantibodies in platelet eluate and plasma were detected by a modified monoclonal antibody immobilization of platelet antigens assay (MAIPA).

RESULTSThe overall positive rate of specific autoantibodies against platelet GPIIb/IIIa and GPIb/IX in plasma was 38.89% and in eluated platelet membrane was 68.52%. The difference between them was significant (corrected chi(2) = 19.39, P < 0.005). The proportion of positive MAIPA results between primary AITP and secondary AITP was not significantly different. There was a significant inverse correlation between antibody level and platelet count.

CONCLUSIONDetection of eluated GP-specific autoantibodies by MAIPA is highly specific and much more sensitive as compared with the measurement of their plasma counterparts in the diagnosing and therapeutic monitoring of AITP.

Adolescent ; Adult ; Autoantibodies ; blood ; Blood Platelets ; immunology ; Female ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Platelet Glycoprotein GPIb-IX Complex ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; immunology

This study was purposed to investigate the expression of RPL36A (ribosomal protein 36a) in the newly diagnosed acute myeloid leukemia (AML) cells and its mechanism at the molecular level. The RPL36A mRNA expression in the newly diagnosed AML cells, U937 cells and normal MNCs was determined by RT-PCR. Small interfering RNA (siRNA) targeting to RPL36A was transfected into U937 cells by Lipofectamine 2000 system. Proliferation, cell cycle, apoptosis of U937 were observed through MTT assay, flow cytometry, acridine orange/ethidium bromide (AO/EB) double staining, TUNEL and Annexin V/FITC respectively. RPL36A mRNA and protein expression levels were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis respectively. The results showed that RPL36A expression in the newly diagnosed AML cells and U937 cells was significantly upregulated. The average OD value of U937 cells transfected with RPL36A siRNA was significantly lower as compared with 3 control groups. The cell percentage in G2-and S-phase increased, which indicated the inhibition effect of RPL36A siRNA on cell proliferation. Remarkable cell apoptosis in U937 cells treated with RPL36A siRNA was observed by AO/EB, TUNEL analysis and Annexin V/FITC assay; RPL36A mRNA and protein expression level of U937 cells treated with siRNA were significantly declined in a time-dependent manner (r=0.9813 and 0.9537). It is concluded that the RPL36A expression in the AML cells is significantly enhanced and the RPL36A gene may be involved in regulation of cell cycle and cell apoptosis of AML, which promotes proliferation of AML cells and inhibits apoptosis of cells.
Adolescent ; Adult ; Aged ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Female ; Humans ; Leukemia, Myeloid, Acute ; pathology ; Male ; Middle Aged ; RNA Interference ; RNA, Small Interfering ; genetics ; pharmacology ; Ribosomal Proteins ; genetics ; pharmacology ; U937 Cells ; Young Adult

OBJECTIVETo report the operative techniques and clinical results of specially designed sural neurocutaneous vascular flap pedicled on a dominant perforator (the diameter > or = 0.8 mm) of the peroneal artery for coverage of soft tissue defects overlying the Achilles tendon.

METHODSAn approximately rectangular sural neurocutaneous vascular flap pedicled on the lowest dominant perforator arising from the peroneal artery was designed and harvested to repair defects over the Achilles tendon. The pedicle was located at a certain part of the flap, which divided the flap into the distal and the proximal parts. After the tendon was repaired, the flap was rotated 180 degrees based on the perforator and the position of the distal and proximal parts of the flap was changed to cover the defects and part of the donor site respectively. In most cases, skin graft was not needed.

RESULTSThe modified flaps were applied in 15 cases. All flaps (ranged from 13 cm x 15 cm - 18 cm x 9 cm ) were transplanted successfully without necrosis, and no vascular problems occurred. Following up for 10-17 months showed both satisfactory functional and cosmetic results.

CONCLUSIONSThe modified flap has reliable blood supply and the special design provides nearly normal outline of the ankle which favorites shoe wearing. It' s an excellent option for covering defects overlying the Achilles tendon.

Achilles Tendon ; injuries ; surgery ; Adolescent ; Adult ; Arteries ; transplantation ; Child ; Female ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Sural Nerve ; blood supply ; Surgical Flaps ; blood supply ; innervation ; Wound Healing ; Young Adult

The purpose of this study was to evaluate whether the DNA vaccine containing idiotypic gene fragment of human B-cell lymphoma cell line Namalwa could elicit the specific anti-idiotypic immune response in vivo. The candidate gene fragment of the lymphoma cell, variable region of heavy chain (VH) of the membranous immunoglobulin, was amplified using Ig superfamily primers by means of RT-PCR. Also, the intact cDNA of murine monocyte chemoattractant protein (MCP-3) was cloned and used as the adjuvant molecular. The two gene fragments of VH and MCP-3 were fused together by 8aa linker peptide with recombinant PCR. Subsequently, the fusion gene fragment was cloned into eukaryonic expression vector pcDNA3.1 to construct DNA vaccine plasmid. Prior to the immunization, the transient transfection coupled with RT-PCR was performed to prove that the recombinant plasmid could express in eukaryonic cells in right way. Then two groups of mice were immunized by intramuscular injection with DNA vaccine and mock plasmid pcDNA3.1 respectively. Three times of injection were performed with 100 micro g plasmid respectively at the beginning of the experiment and 2, 4 weeks after the first injection for all the groups. FACS analysis was chosen to detect the antibodies recognizing lymphoma cells at different time following vaccination. The results demonstrated that specific anti-idiotypic antibody could be detected in the group of DNA vaccine immunized mice as early as eight weeks after the first immunization. Further test demonstrated that the anti-idiotypic antibody could maintain for at least twenty weeks with high titer. Anti-idiotypic antibodies were elicited in three of five mice of the DNA vaccine immunized group. The Abs of DNA vaccine immunized mice could only recognize Namalwa cell line instead of another unrelated human cell line A549. There is no cellular response detected in the DNA vaccine immunized mice. It is concluded that the DNA vaccine containing fused MCP3-VH sequence could elicit specific anti-idiotypic antibody against B-cell lymphoma in vivo and could be used in further study of DNA vaccine against B-cell lymphoma. The results would provide the basis for further studies and optimization of this therapeutic strategy on patients with B-lymphoproliferative disease.
Animals ; Antibodies, Anti-Idiotypic ; blood ; COS Cells ; Cancer Vaccines ; immunology ; Chemokine CCL7 ; Cytokines ; Humans ; Immunization ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Lymphoma, B-Cell ; immunology ; Mice ; Mice, Inbred BALB C ; Monocyte Chemoattractant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; immunology

OBJECTIVETo investigate the features of circulating plasmacytoid dendritic cells (pDCs) in patients with chronic hepatitis B for a better understanding of the immunopathogenesis of HBV infection.

METHODSFresh blood samples were collected from 20 patients with chronic hepatitis B (CHB) and from 15 healthy individuals who served as controls. pDCs were isolated from peripheral blood mononuclear cells (PBMCs) using immunomagnetic assay and detected by flow cytometry. Fresh PBMCs and isolated pDCs were stimulated in vitro using CpG ODN2216. The supernatants were measured for IFNa production using ELISA.

RESULTSThe peripheral pDCs frequency in CHB patients (0.192%+/-0.110%) was markedly lower than that in the healthy controls (0.287%+/-0.142%). After being pulsed with CpG ODN2216, the isolated pDCs produced lower levels of IFNa and expressed lower levels of CD80 and CD40 in the CHB patients when compared to those of the healthy controls. The level of IFNa was (972.6+/-705.5) pg/ml in the patients and (3 142.9+/-1 292.2) pg/ml in the controls. Moreover, the pDCs frequency was reversely correlated with serum ALT levels in these HBV infected patients.

CONCLUSIONThe reduced number and impaired function of circulating pDCs in patients with CHB may be related to their disease progression.

Adult ; Case-Control Studies ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; Female ; Flow Cytometry ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Interferon-alpha ; biosynthesis ; Male ; Middle Aged ; Oligodeoxyribonucleotides ; pharmacology

AIM To observe the clinical effects and safety of Zibei Zhike Granules (Asteris Radix et Rhizoma,Ardisiae japonicae Herba,Fritiliariae cirrhosae Bulbus,etc.) for acute broncho-bronchitis with remained toxicity lingering lung.METHODS A multi-center,randomized,double-blinded,double-simulation and positive drug parallel controlled trial was adopted.Two hundred and forty cases of patients with the 1 ∶ 1 ratio were assigned to treatment and control group.The treatment group were treated with Zibei Zhike Granules and the control group were treated with Zikebao Tablets (Asteris Radix et Rhizoma,Citri rubrum exocarpium,Platycodi Radix,etc.).The treatment course lasted five days.RESULTS The total effective rate of acute broncho-bronchitis in the treatment group was 73.04％,and 54.78％ in the control group.There was statistical significance between the total effective rate of the two groups (P ＜ 0.01).The total effective rate of traditional Chinese medicine syndrome in the treatment group was 73.91％ and 60.86％ in the control group,and there was statistical significance between the total effective rate of the two groups (P ＜ 0.05).The treatment group showed better clinical effects in improving individual symptoms of cough and spitting sputum.CONCLUSION Zibei Zhike Granules has good clinical effects on acute broncho-bronchitis.