Objective:To validate the usefulness of copy number variation sequencing (CNV-seq) in detecting the deletion/duplication of the DMD gene in Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) patients. Methods:One hundred and seventy-seven cases who visited the Department of Medical Genetics, Affiliated Hospital of Kunming University of Science and Technology/the First People′s Hospital of Yunnan Province from April 2018 to November 2023 were collected. All patients had previously accepted multiplex ligation-dependent probe amplification (MLPA) to detect the deletion/duplication of the DMD gene, including 90 cases of normal control with a negative result of MLPA and 87 cases with the deletion or duplication of the DMD gene (61 cases of DMD and 26 cases of BMD). CNV-seq was performed in a single-blind manner to detect DMD gene deletion or duplication for all of 177 cases to obtain the detection efficiency of CNV-seq in comparison with MLPA. Results:Comparing to MLPA, CNV-seq had a coincidence rate of 88.7% (157/177) for detecting DMD gene deletion/duplication, with a sensitivity of 77.0% (67/87), a specificity and a positive predictive value of both 100.0% (90/90 and 67/67, respectively), a negative predictive value of 81.8% (90/110), and a Kappa value of 0.773. Of the 87 patients with the deletion or duplication of the DMD gene, CNV-seq detected 67 cases with DMD gene deletion/duplication, including 62 cases with deletion and 5 cases with duplication, with fragment ranging from 150 to 750 kb. While CNV-seq missed 23.0% (20/87) of positive cases, mainly due to the involved fragments spanning only 1 to 4 exons, and with a variation size less than 50 kb, below the resolution (100 kb) of CNV-seq. The detection rate of CNV-seq in BMD cases (84.6%, 22/26) was a little higher than that in DMD cases (73.8%, 45/61), but there was no significant difference between 2 subgroups ( χ2=1.211, P=0.271). The results of CNV-seq in normal controls were all negative, and consistent with the results of MLPA. Conclusion:CNV-seq can detect 77.0% (67/87) of deletion/duplication of the DMD gene in patients with DMD/BMD, while the deletion/duplication less than 100 kb may be inevitably unidentified, therefore it is recommended as an assistant screening technique in prenatal diagnosis for DMD gene deletion or duplication.

Objective:To validate the usefulness of copy number variation sequencing (CNV-seq) in detecting the deletion/duplication of the DMD gene in Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) patients. Methods:One hundred and seventy-seven cases who visited the Department of Medical Genetics, Affiliated Hospital of Kunming University of Science and Technology/the First People′s Hospital of Yunnan Province from April 2018 to November 2023 were collected. All patients had previously accepted multiplex ligation-dependent probe amplification (MLPA) to detect the deletion/duplication of the DMD gene, including 90 cases of normal control with a negative result of MLPA and 87 cases with the deletion or duplication of the DMD gene (61 cases of DMD and 26 cases of BMD). CNV-seq was performed in a single-blind manner to detect DMD gene deletion or duplication for all of 177 cases to obtain the detection efficiency of CNV-seq in comparison with MLPA. Results:Comparing to MLPA, CNV-seq had a coincidence rate of 88.7% (157/177) for detecting DMD gene deletion/duplication, with a sensitivity of 77.0% (67/87), a specificity and a positive predictive value of both 100.0% (90/90 and 67/67, respectively), a negative predictive value of 81.8% (90/110), and a Kappa value of 0.773. Of the 87 patients with the deletion or duplication of the DMD gene, CNV-seq detected 67 cases with DMD gene deletion/duplication, including 62 cases with deletion and 5 cases with duplication, with fragment ranging from 150 to 750 kb. While CNV-seq missed 23.0% (20/87) of positive cases, mainly due to the involved fragments spanning only 1 to 4 exons, and with a variation size less than 50 kb, below the resolution (100 kb) of CNV-seq. The detection rate of CNV-seq in BMD cases (84.6%, 22/26) was a little higher than that in DMD cases (73.8%, 45/61), but there was no significant difference between 2 subgroups ( χ2=1.211, P=0.271). The results of CNV-seq in normal controls were all negative, and consistent with the results of MLPA. Conclusion:CNV-seq can detect 77.0% (67/87) of deletion/duplication of the DMD gene in patients with DMD/BMD, while the deletion/duplication less than 100 kb may be inevitably unidentified, therefore it is recommended as an assistant screening technique in prenatal diagnosis for DMD gene deletion or duplication.

Objective To investigate the severity of allogenic rejection after partial liver transplantation (PLTx) with different graft weight to recipient liver weight (GW/RLW).Methods The full-size liver transplantation (group A),GW/RLW ＞33％ PLTx (group B) and GW/RLW ＜ 30％ PLTx (group C) were set up using BN rats and Lewis rats as donors and recipients,respectively.All recipients were observed for 28 days.The Banff RAI grading,survival rate,jaundice and body weight recovery were evaluated to determine the severity of acute allogeneic rejection.Two PLTx groups,group B1 (GW/RLW＞33％) and group C1 (GW/RLW＜30％),were established to assess the mRNA level of IL-2,GranzymeB,Perforin and CD3 48 h and 7 days postoperatively.Additionally,the mRNA level of B7-H1,the ratio of Ki67 + hepatocytes and the liver enzymes were also assessed 7 days postoperatively.Results All recipients in group C died within 22 days postoperatively,presenting with severe lymphocytic infiltration and vascular endothelialitis.All recipients in group A and group B survived until the end of observation time.All recipients in group A survived and presented with a mild lymphocytic infiltration and rare vascular endothelialitis.Group B presented with moderate lymphocytic infiltration and moderate vascular endothelialitis.The Banff RAI grading in group C was significantly higher than that in group A and group B (P ＜ 0.05).In accordance with the result of histology and survival rate,group B and group C presented with earlier jaundice and lower body weight recovery than that of group A (P＜0.05).As compared with group B1,group C1 presented with higher mRNA levels of Perforin,GranzymeB,IL-2 and CD3,higher level of liver enzymes and heavier liver graft weights.Besides,the mRNA level of immunosuppresive molecule B7-H1 in group C1 was lower than that of group B1.However,there was no significant difference in the ratio of Ki67 + hepatocytes between group B1 and group C1.Conclusion The allogenic liver rejection may be enhanced by reducing the GW/RLW.

Objective To investigate the effectiveness of Varenicline combined with MEDCHART application software for health education and follow-up in patients at smoking cessation clinic.Methods A total of 104 cases who had smoking cessation counseling and intervention at smoking cessation clinic from February 2014 to January 2015 were divided randomly into MEDCHART APP group (n=51) and conventional group (n=53).Both groups were intervened by brief admonishment and given Varenicline.The 9-12 weeks and 26th week protracted continuous abstinence rates were observed.Results There were no significant difference in 9-12 week protracted withdrawal rate between 26th week APP and conventional groups (56.9％ vs.45.3％),the 26th week abstinence rates in group MEDCHART APP (41.2％) were significantly higher than the conventional group (20.8％)(x2=5.088,P＜0.05).There was significant difference in full course completion rates between the two groups;15 cases (29.4％) completed the course in group MEDCHART APP,whereas 5 cases (9.4％) did so in conventional group (x2=4.405,P＜0.05).Conclusion MEDCHART APP can strengthen health education and follow-up,which helps extend the time for Varenicline,increase 26th week smoking cessation withdrawal rate.It is a useful tool for smoking cessation education and follow-up.

Objective To study antibacterial effect of formulas, borneol, honey, gentamicin in wet honey ice bedsores.Methods The third-stage infective bedsore model in rabbit were estabished with Staphylococcus aureus ( S.aureus) , Escherichia coli ( E.coli) , and Pseudomonas aeruginosa ( P. aeruginosa).The rabbits with bedsore were respectively compressed with gauze of honey+borneol, honey+gentamicin,borneol+gentamicin,borneol+gentamicin+honey, vaseline as control group.The strain identification and colony counts were observed before and after treatment.ResuIts The colony count of S.aureus, E.coli and P.aeruginosa in honey+borneol, honey+gentamicin, borneol+gentamicin, borneol+gentamicin+honey post-treatment was significantly higher than that of pre-treatment, respectively (P<0.05).The colony count of three strains in above four formulas post-treatment was significantly lower than that in vaseline group, respectively (P <0.05).The colony count of three strains in borneol +gentamicin +honey (original formula) post-treatment was significantly lower than that in the other formulas, respectively (P<0.05).ConcIusion The antibacterial effect of borneol+gentamicin+honey (original formula) is the best.

Objective To study the antibacterial effect of borneol, honey, gentamicin in Ruchuang Bingmi hydropathic compress agent.Method Staphylococcus aureus(S.aureus), Escherichia coli(E.coli), Pseudomonas aeruginosa(P.aeruginosa) were used to produce the third phase of infection decubitus animal models, respectively.Vaseline as the control group, divided into 12 groups:Vaseline-S.aureus, Vaseline-E.coli, Vaseline-P.aeruginosa;Honey-S.aureus,Honey-E.coli,Honey-P.aeruginosa;Borneol -S.aureus;Borneol -E.coli;Borneol-P.aeruginosa;Gentamicin-S.aureus;Gentamicin-E.coli;Gentamicin-P.aeruginosa;6 rabbits in each group.Honey, borneol, gentamicin was made into a gauze in treatment for decubitus .Organizations do strain identification and colony counts was observed before and after taking the treatment.ResuIts Borneol group ( F =11.059,P<0.01).,there was differences of each groups count cultured by borneol;Time ×Strains(F=11.281,P=0.009),there was no significant interaction between the two groups;gentamicin(F=7.99,P=0.000),gentamicin culture showed a difference in each group count;Time ×Strains(F=12.531, P<0.07),interaction between the two groups showed significant.Borneol has no antibacterial effects on P.aeruginosa , had a certain antibacterial activityon S.aureus and E coli;gentamicin had good antibacterial effect on the three kinds of bacteria, and against P.aeruginosa was particularly significant.ConcIusion The antibacterial activity of gentamicin is better than single herbs borneol in Ruchuang Bingmi hydropathic compress agent, honey has no antibacterial effect.

OBJECTIVETo detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.

METHODSRT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.

RESULTSSignificant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).

CONCLUSIONCXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokines, CXC ; metabolism ; Humans ; Osteosarcoma ; metabolism ; pathology ; Prognosis

Objective To investigate treatment and prognosis of patients with osteosarcoma of the extremities.Methods A total of 311 patients with osteosarcoma of the extremities,who had undergone treatment in our institute from 1998 to 2008,were enrolled in this retrospective study.Kaplan-Meier survival curve and Cox regression model were used to analyze the correlation between survival rate and variables including patients' demographics,chemotherapy,surgery,complications,and tumor metastasis.Results Among 311 patients,there were 206 males and 105 females,aged from 5 to 56 years (average,18.6 years).A total of 282 patients underwent aggressive or radical surgery,including 149 cases of limb salvage surgery and 133 cases of amputation surgery.One hundred and five patients underwent standard chemotherapy and 206 patients underwent non-standard chemotherapy.The 5-year survival rate was 57.4％ in patients treated with standard chemotherapy,36.3％ in patients treated with non-standard chemotherapy,16.8％ in patients with lung metastasis,50.7％ in patients without lung metastasis,56.6％ in patients who underwent limb salvage surgery,31.8％ in patients who underwent amputation surgery,44.6％ in patients with Enneking stage Ⅱ B and 33.1％ in patients with Enneking stage Ⅲ.For patients treated by amputation surgery,because non-standard chemotherapy which was performed in most of them and other confounding factors,the 5-year survival rate of them was lower.The Cox regression analysis showed that lung metastasis and non-standard chemotherapy were associated with inferior outcomes.Conclusion Neoadjuvant chemotherapy combined with aggressive or radical surgery could cure about 60％ of patients with osteosarcoma of the extremities.Lung metastasis and non-standard chemotherapy are risk factors that severely affect prognosis.

Taking the national key scientific and technological project in 11th Five-year and 10th Five-year Plan as its main clue,the innovative Chinese materia medica(CMM)research course has been reviewed in this paper.Based on the enlightenment from the first herbal medicine VeregenTM,which has been approved by FDA,together with the study and comprehension of many great achievements of new drug research,the strategy of innovative CMM research and development(R&D)and its practical route is going to be put forward in this article.With the innovative Compound Prescription of CMM research as the main innovative approach,innovation can be realized by the following three aspects:New substances,including new compound,new effective fraction/group or new components and their combinations;new indication of known compounds or extracts;the secondary development of high-quality Chinese patent medicine.Besides,in order to support the R & D of original drugs with Chinese characteristics,the international experiences in target-dependent screening and evaluation and optimization of leading compounds based on clinical application should be referenced,and the mechanism and targets of traditional Chinese medicine by genetic and molecular level be clarified.