Abstract: Objective　Quantitative assessment of risk factors of clonorchiasis can provide prevention for clonorchiasis. Methods　Articles were retrieved in Chinese and English electronic databases from January 1, 2000 to December 31, 2020, including Wanfang Data, CNKI, PubMed, Web of Science, Embase. All studies were screened based on inclusion and exclusion criteria, and the quality of all enrolled literatures was evaluated. The software RevMan version 5.3 was used for Meta-analysis. The heterogeneity, sensitivity and publication bias of all included studies were analyzed. Results　A total of 95 articles were retrieved, and 6 were included in this Meta-analysis which were case-control studies. There were 5 articles in Chinese and 1 in English. There was no single literature with a large impact on the results, and the results of this study were relatively stable. There were 1 170 cases of clonorchiasis in total and 1 291 cases in control. Most of the cases were from hospital patients and community residents, and the floating population was small. Meta-analysis showed that there were three independent risk factors: raw or semi-raw fish, raw or semi-raw shrimp, mix raw and cooked cutting boards, with a combined OR (95%CI) of 2.32(1.86, 2.88), 3.99(2.42, 6.58), 2.18(1.51, 3.14), respectively, with low heterogeneity consistent with the results of the total sample study: I2 values for risk factors were 30%, 12%, 27%, respectively. The results of bias tests showed no publication bias (P=0.731, 0.725, 0.334, P>0.05). Conclusions　The key risk factors of clonorchiasis are raw or semi-raw fish, raw or semi-raw shrimp, mix raw and cooked cutting boards. Guidance and health education should be strengthened. It is necessary to strengthen the surveillance of clonorchiasis in the floating population, such as traveler, businessman and student.

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.

OBJECTIVETo study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.

METHODSE. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.

RESULTSA total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.

CONCLUSIONCTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.

Bacteremia ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; genetics ; isolation & purification ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Genotype ; Hospitalization ; statistics & numerical data ; Humans ; Liver Cirrhosis ; therapy ; Microbial Sensitivity Tests ; beta-Lactamases ; genetics ; metabolism

OBJECTIVETo evaluate clinical efficacy of suture anchors in treating acute injuries of medial collateral ligament (MCL) of knee at degree III.

METHODSTwenty-seven patients with degree III acute MCL injuries of knee were treated with suture anchors from January 2007 to June 2011. There were 15 males and 12 females, aged from 19 to 56 (averaged 32.6) years old. The time from injury to operation was 3 to 10 days, averaged 6 days. Symptoms and physical signs before and after treatment were observed, Lysholm scoring were used to evluated clinical efficacy.

RESULTSAll patients were followed up from 16 to 30 months with an average of 21.6 months. The stability of knee joints was good in all patients. Abduction stress test was negative when the knee joint was straightened at 0 degrees and flexed at 30 degrees. The average degree of flexed knee (67.00 +/- 5.80) degrees preoperatively was lower than that of postoperatively (136.50 +/- 6.30) degrees at 1 year. According to Lysholm scoring, preoperative scores ranged from 30 to 43 points, averaged 36.46 +/- 1.48; 1 year after operation ranged from 87 to 100 with an average of 91.50 +/- 3.80 and higher than postoperative. Twenty patients got an excellent results, 5 good and 2 fair.

CONCLUSIONSuture anchors in treating acute injuries of medial collateral ligament of knee at degree III has following advantages: small range of tissue dissection, easy to operate, reliable fixation and less complications.

Acute Disease ; Adult ; Collateral Ligaments ; injuries ; surgery ; Female ; Humans ; Knee Injuries ; surgery ; Male ; Middle Aged ; Suture Anchors

Objective To evaluate X-ray,CT and MRI findings of soft tissue infections in AIDS. Methods Three cases of soft tissue infections with AIDS were retrospectively analyzed by comparing the imaging findings with pathological results.All patients were performed MRI,X-ray was in 1 case,CT was in 1 case.Results Cellulitis was in 1 case:MRI showed extended thickening of subcutaneous tissues, ill-defined hypointense areas on T_1WI and hyperintensity on T_2WI,and reticular pattern on GRE. Necrotizing fasciitis was in 1 case:MRI showed obvious thickening of subcutaneous tissues and deep fasciae, abnormally increased signal intensity on T_1 and T_2WI.Fluid collections were within muscles and muscles interval on fat-suppressed T2 WI.Tuberculosis was in 1 case:CT demonstrated multiple low density areas in the subcutaneous tissues and clear peripheral rim enhancement.MRI appeared hypointense on T_1WI and hyperintensity on T_2WI,and peripheral rim enhancement following gadolinium injection.Conclusion Infections of soft tissue are common complication in patients with AIDS,radiology is important in early diagnosis and treatment planning in this population.

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

The study was aimed to evaluate if the modified in situ cryopreservation could affect the biological function of mesenchymal stem cells (MSC) in vitro. Mesenchymal stem cells from human bone marrow were isolated by standard method and characterized with their morphology, cell-surface antigen profile and differentiation repertoire in vitro. The culture-expanded MSC were cryopreserved in situ with culture medium (DMEM-LG) containing 10% D MSO and 30% selected FCS in -70 degrees C. Following recovery of cryopreservation, differentiation to adipocytes, chondrocytes, and osteoblast in vitro and cell cycle analysis were performed to investigate whether the cryopreservation would change the differentiation potential of MSC. The results showed that after recovery of cryopreservation, there was no changes detected as compared with the culture-expanded MSC in both differentiation potency and growth pattern at 12 weeks. In conclusions: this optimized short term in situ cryopreservation at -70 degrees C could retain biological characteristics of human MSC for at least 3 months, and this method may be useful for cryopreservation of hum an bone marrow MSCs.
Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Separation ; Cell Survival ; Cryopreservation ; Humans ; Mesenchymal Stromal Cells ; cytology

As main component of fetal liver hematopoietic microenvironment, different stromal cells may play distinct roles in the regulation of hematopoietic stem cell self-renewal, proliferation and differentiation. It is a unique approach to establish stromal cell lines for analyzing the interaction of hematopoietic cells with the stroma on the clonal level to dissect the function of hematopoietic microenvironment. In this study two immortal stromal cell lines-A4, B3 were established from mouse embryonic day 12.5 fetal liver by transfection of pSV3 neo plasmid. A4 exhibited a fibroblast-like morphology, 25 hours population doubling time as well as high levels of CD29, CD44, UEA-1 and low levels of CD105 expression. In contrast B3 displayed an epithelium-like morphology, 37 hours population doubling time along with high levels of CD105 and low levels of UEA-1 expression. In addition no or low levels of CD31, CD34, CD45 and CD144 expression were found in the two cell lines. These results indicate that A4, B3 are two discriminating cell lines in terms of morphological characters, growth behaviors and surface molecular expression types. Next functional assays using Limited-Diluted Assay(LDA) and Bulk-LTC-IC were done: both stromal cell lines had similar ability to maintain the survival proportion of inoculated mouse bone marrow-derived Long-Term Culture-Initiating Cells (LTC-ICs), and they could also support LTC-ICs expansion up to 4 weeks by co-culture in vitro. More strikingly, B3 could expand the absolute number of LTC-ICs over 13-fold at week 4 than that of week 0, and the ability of B3 to expand absolute number of LTC-ICs was over 8-fold of that of A4. Proportions of LTC-ICs in proliferating cell populations in two long-term culture systems was similar at week 4, no matter with or without extra cytokines. Further study indicated that the ability of LTC-ICs to yield CFCs was held as the number 6( +/- 1.2) after 4 weeks co-culture. Extra cytokines-SCF + IL-3 + IL-6 + Epo had no influence in the maintenance and expansion of LTC-ICs, but expanded the absolute number of CFCs and proliferating cell populations, and maintained similar proportions of LTC-ICs and CFCs in the end in two culture systems at week 4. Take together, these results implicate that B3 may act as an important functional component in embryonic day 12.5 fetal liver microenvironment to effectively expand primitive hematopoietic cells, yield more committed hematopoietic progenitor cells and mature hematopoietic cells to meet the need of quick development especially in the early phase of embryonic development. Alternatively A4 can probably function as being a structural element. Moreover the function of hematopoiesis-associated cytokines employed in this investigation was not to expand LTC-ICs, but to modulate the limited-proliferation and differentiation of CFCs. The maintenance and proliferation of LTC-ICs may depend on the type of the stromal cell as well as the interaction between stromal cells and LTC-ICs. It is suggested that B3 with cytokines including SCF, IL-3, IL-6, EPO in vitro can mimic embryonic day 12.5 hematopoietic microenvironment to investigate the mechanism of interaction between hematopoietic microenvironment and hematopoietic cells in clonal level.
Animals ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cell Culture Techniques ; methods ; Female ; Fetus ; cytology ; Hematopoietic Stem Cells ; cytology ; metabolism ; Hyaluronan Receptors ; metabolism ; Integrin beta1 ; metabolism ; Interleukin-3 ; metabolism ; Interleukin-6 ; metabolism ; Leukocyte Common Antigens ; metabolism ; Liver ; embryology ; Male ; Mice ; Mice, Inbred C57BL ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Stromal Cells ; cytology ; metabolism

Mesenchymal stem cells (MSCs), precursors of diverse stromal cells, can support hematopoiesis in vitro and can promote the implantation of hematopoietic stem cells in vivo when co-transplanted with CD34(+) cells. The aim of this study was to investigate the potential effect of MSCs on the hematopoietic development of embryonic stem cells (ES cells) and the feasibility of a novel system in which ES cells will be co-cultured with MSCs. The murine bone marrow MSCs were isolated and cultured and then their phenotype and differentiation function were identified with FCM and histochemical technique. The CCE cells, murine ES cell line, were co-cultured with the isolated MSCs and the hematopoietic differentiation of CCE cells was observed with hematopoietic clonogenic assay and RT-PCR. The results showed that the morphology of MSCs became gradually homogeneous with the passage culture of cells. After passage 4, the marker of Sca-1, CD29, CD44 and CD105 were highly expressed, however, CD34 and CD45, the specific marker of hematopoietic and endothelial cells, could hardly be identified. The isolated MSCs differentiated into adipocytes and osteoblasts in specific induction culture system. After maintaining culture on mouse embryonic fibroblasts, CCE cells were plated in suspended culture system with only differentiation inductive agents and co-culture system in which MSCs were added. Compared with CCE cell suspended culture, the cells differentiated into embryoid body were obviously enhanced and there were no colony-forming cells in the co-culture system of ES cells and MSCs. In addition, transcription factor Oct-4 in co-cultured CCE cells was expressed and hematopoietic markers, Flk-1, GATA-1 and beta-H1, were negative. The ability of embryoid bodies derived from the co-culture system to produce hematopoietic colonies was markedly higher than that from the suspended culture system. It is concluded that MSCs inhibit the initial differentiation of ESC and enhance hematopoietic differentiation ability of the co-cultured ES cells.
Animals ; Bone Marrow Cells ; physiology ; Cell Differentiation ; Coculture Techniques ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroid-Specific DNA-Binding Factors ; Female ; GATA1 Transcription Factor ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mesenchymal Stromal Cells ; physiology ; Mice ; Mice, Inbred C57BL ; Octamer Transcription Factor-3 ; Transcription Factors ; genetics

OBJECTIVETo study the status of beta-lactamase produced by multiresistant Aeromonas selected from cirrhosis patients to provide reference for treatment and reduce resistance and control spreading.

METHODSFour multiresistant Aeromonas strains isolated from serious liver cirrhosis patients from the No. 302 hospital. The TEM resistant genes were detected by PCR and agarose gel electrophoresis.

RESULTSThree TEM-1 positive strains were detected from four multiresistant Aeromonas isolates consisting of one Aeromonas sobria and three Aeromonas hydrophila isolated from blood and ascites. This was further confirmed by gene sequencing. The multiresistance to antibiotics was higher in four Aeromonas isolates. All strains tested were resistant to ampicillin, cefazolin and cefmetazole.The cirrhosis patients who suffered from Aeromonas infection had poor prognosis and had mortality rate of 3/4.

CONCLUSIONThe beta-lactamase TEM-1 resistant genes was detected in clinical multiresistant Aeromonas strain isolated from serious cirrhosis patients.The results suggested that TEM-1 was the main resistance mechanism of Aeromonas strain and was reduced by adding enzyme inhibitor.

Adult ; Aeromonas ; drug effects ; genetics ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Gram-Negative Bacterial Infections ; microbiology ; Humans ; Liver Cirrhosis ; microbiology ; Male ; Middle Aged ; Prognosis ; beta-Lactamases ; genetics