The clinical value of combined detection of serum angiopoietin 2 and Clara cell protein 16 in the early diagnosis of acute respiratory distress syndrome / 中华急诊医学杂志

Objective@#To investigate the clinical value of combined detection of serum angiopoietin 2 (Ang-2) and Clara cell protein 16 (CC16) in the early diagnosis of acute respiratory distress syndrome (ARDS).@*Methods@#Two hundred critical patients, treated at the Department of Critical Care Medicine, Bao'an District People's Hospital, Shenzhen during March 2015 and September 2016,were included in the study. According to the Berlin standard, patients were divided into two groups (n=100 each group): the ARDS group and non-ARDS group.The serum levels of Ang-2 and CC16 were measured by enzyme-linked immunosorbent assay (ELISA) on the first and second day of admission for each patient, in addition to completing APACHEⅡ score, medical history, vital signs collection and other biochemical indicators detection. Finally, paired-samples t test was used to analyze the data. The multiple ROC curve was used to calculate the area under the curve of the reference index of the serum levels of Ang-2 and CC16 on the first and second day.@*Results@#On the first and second day, the serum levels of Ang-2 and CC16 were significantly higher in ARDS patients than those in non-ARDS patients, and there was a correlation between the serum levels of Ang-2 and CC16 in ARDS patients. The ROC curve showed that the combined detection of Ang-2 and CC16 on the first day achieved a highest sensitivity of 75.9% and detection of CC16 on the first day achieved a highest specificity of 70.2%.@*Conclusion@#Single-detection of serum levels of Ang-2 and CC16 could be used for early diagnosis of ARDS, and the combined detection of both has a higher sensitivity than single detection.

Preliminary experience in guiding individualized targeted therapy of advanced metastatic renal cell carcinoma with gene detection technology / 中华泌尿外科杂志

Objective To explore the efficacy and tolerance of adverse reactions of gene detection technique in guiding individualized targeted therapy for advanced metastatic renal cell carcinoma.Methods Retrospective analysis was performed on the clinical data of 62 patients with advanced metastatic renal cell carcinoma before and after receiving targeted drug treatment in our department from October 2015 to October 2017.Among the 62 patients,there were 36 males and 26 females,with an average age of (54 ± 13) years old.16 patients were treated with sunitinib,20 patients were treated with sorafenib and 26 patients were treated with pazopanib.A total of 28 patients (individualized group) were selected to receive targeted drug according to the results of gene detection,and 34 patients were treated with targeted drug empirically (empirical group).In individualized group,there were 17 males and 11 females with the average age of (51.3 ± 15.6) years old.20 patients accepted the operation.The distant metastasis included bone metastasis in 21 cases,lung metastasis in 7 cases,liver metastasis in 16 cases,epidermal metastasis in 4 cases and lymphatic metastasis in 14 cases.According to risk of MSKCC,the case number of low risk,moderate risk and high risk were 15,7,6,respectively.7 patients were treated with sunitinib,8 patients were treated with sorafenib and 13 patients were treated with pazopanib.In empirical group,there were 19 males and 15 females with the average age of (56.3 ± 10.1) years old.22 patients accepted the operation.The distant metastasis included bone metastasis in 20 cases,lung metastasis in 5 cases,liver metastasis in 13 cases,epidermal metastasis in 3 cases and lymphatic metastasis in 15 cases.According to risk of MSKCC,the case number of low risk,moderate risk and high risk were 20,g,6,respectively.9 patients were treated with sunitinib,12 patients were treated with sorafenib and 13 patients were treated with pazopanib.The baseline characteristics of the two groups of patients,including gender,age,whether operation was performed,site of metastasis,and risk of MSKCC,didn't show significant difference.Patients in both groups received the standard treatment regimen and the follow-up duration was 4-26 months to observe the efficacy,progression-free survival and tolerance to adverse reactions of the targeted therapy.Results After 12 months of treatment,15 patients in the individualized group was recorded objective remission.7 patients in the empirical group was recorded objective remission,as well.The tumor control efficacy of the individualized group was significantly better than that of the empirical group (46.4％ vs.20.6％,P =0.03).Meanwhile,the median progression-free survival time (15.2 months,3.7-24.2 months) in the individualized group was significantly longer than that in the empirical group (12.1 months,2.8-22.1 months) (P =0.009).Compared with the empirical group,the higher incidence of targeted treatment-related adverse reactions occurred in the individualized group,including thrombocytopenia (46.4％ vs.17.6％ P =0.014),leukopenia (46.4％ vs.17.6％ P =0.005),hypertension (71.4％ vs.44.1％,P =0.031) and hypothyroidism(60.7％ vs.29.4％,P=0.013).Conclusions Compared with the patients with empirical drugs,the application of gene detection technique to select individualized targeted drugs for the treatment of advanced metastatic renal cancer is obvious curatively effective,and to a certain extent extends the progression-free survival time of patients.

Clinical application of 3D printing combined with 3D laparoscopy in partial nephron-sparing surgery for partial endogenous renal cell carcinoma / 中华泌尿外科杂志

Objective To investigate the clinical feasibility and effectiveness of 3-D printing (3DP) combined with 3-D laparoscopic nephron-sparing surgery (LNSS) for partial endogenous renal cell carcinoma.Methods A retrospective analysis was made of the clinical data of 79 patients with partial endogenous renal cell carcinoma who were admitted to our department from July 2015 to October 2018.There were 46 males and 33 females.Their average age was (50.9 ± 7.9) years old,ranged from 33 to 68 years old.Tumor stages were T1aN0M0 in 53 cases and T1bN0M0 in 26 cases.The preoperative serum creatinine ranged from 40 to 107 μmol/L,with an average of (72.4 ± 14.2) μmol/L.The preoperative GFR ranged from 19 to 54 ml/min,with an average of (40.2 ± 6.2) ml/min.Thirty-four patients underwent 2-D laparoscopic nephron-sparing surgery (2DLNSS) based on preoperative enhanced CT scans.Forty-five patients underwent 3-D printing (3DP) based on three-dimensional reconstruction of renal CT scans.Seventeen patients underwent 2-D laparoscopic nephron-sparing surgery guided by 3-D printing model(3DP-2DLNSS),and 28 patients underwent 3-D laparoscopic nephron-sparing surgery guided by 3-D printing (3DP-3DLNSS).Serum creatinine levels ranged from 42 to 122 μmol/L with an average of (86.3 ± 14.8) μmol/L,and creatinine levels ranged from 8 to 66 μmol/L with an average of (19.1 ± 14.1) μmol/L.Six months after operation,the GFR of the kidney was 9-36 ml/min with an average of (21.4 ± 6.4)ml/min,and the fluctuation range was 6-40 ml/min with an average of (19.2 ± 8.8) ml/min.There was no statistical difference in the incidence of complications and pathological types after operation.Results There was no statistical difference in general data of preoperative patients.In intraoperative and post-operative statistics,the time of exploring renal artery was shorter than that of 2DLNSS (33.7 ± 7.5) min in 3DP-2DLNSS (28.3 ± 8.2,P =0.015) min and 3DP-3DLNSS (27.8 ± 6.5,P =0.002) min.In tumor detection time,3 DP-2DLNSS was shorter than 2DLNSS group (41.2 ± 6.6 vs.46.5 ± 6.9 min,P =0.012),and 3 DP-3DLNSS was shorter than 3DP-2DLNSS (35.4 ± 7.3 vs.41.2 ± 6.6 min,P =0.009).In warm ischemia time,3DP-2DLNSS min was shorter than 2DLNSS (23.5 ±9.7 vs.33.9 ±7.5 min P ＜0.001),and 3DP-3DLNSS was shorter than 3DP-2DLNSS (18.3 ± 4.6 vs.23.5 ± 9.7,P =0.023).In surgical time,3DP-2DLNSS (115.7 ± 23.0) min and 3DP-3DLNSS (103.3 ± 22.8) min were shorter than 2DLNSS (132.4 ± 28.9) min (P =0.031,P ＜ 0.001).In intraoperative bleeding volume,3 DP-3 DLNSS was less than 2DLNSS (117.9 ± 17.9 vs.130.6 ± 16.8,P =0.009) ml.Fasting for 1 to 4 days after operation,with an average of (1.7 ± 0.8) days.The indwelling catheterization ranged from 1 to 8 days after operation,with an average of (3.9 ± 1.3) days.Negative pressure drainage was removed 2-9 days after operation,with an average of (4.9 ± 1.4) days.And the hospitalization 5-11 days after operation,with an average of (7.3 ± 1.5) days.Conclusions Preoperative 3D printing combined with intraoperative 3D laparoscopic nephron sparing surgery for partial endogenous renal tumors is safe and effective,which is superior to the previous CT scan alone and intraoperative 2D laparoscopic treatment.

The clinical value of combined detection of serum angiopoietin 2 and Clara cell protein 16 in the early diagnosis of acute respiratory distress syndrome / 中华急诊医学杂志

Objective To investigate the clinical value of combined detection of serum angiopoietin 2 (Ang-2) and Clara cell protein 16 (CC16) in the early diagnosis of acute respiratory distress syndrome (ARDS).Methods Two hundred critical patients, treated at the Department of Critical Care Medicine, Bao'an District People's Hospital, Shenzhen during March 2015 and September 2016,were included in the study. According to the Berlin standard, patients were divided into two groups (n=100 each group): the ARDS group and non-ARDS group.The serum levels of Ang-2 and CC16 were measured by enzyme-linked immunosorbent assay (ELISA) on the first and second day of admission for each patient, in addition to completing APACHEⅡ score, medical history, vital signs collection and other biochemical indicators detection. Finally, paired-samplest test was used to analyze the data. The multiple ROC curve was used to calculate the area under the curve of the reference index of the serum levels of Ang-2 and CC16 on the first and second day.Results On the first and second day, the serum levels of Ang-2 and CC16 were significantly higher in ARDS patients than those in non-ARDS patients, and there was a correlation between the serum levels of Ang-2 and CC16 in ARDS patients. The ROC curve showed that the combined detection of Ang-2 and CC16 on the first day achieved a highest sensitivity of 75.9％ and detection of CC16 on the first day achieved a highest specificity of 70.2％. Conclusion Single-detection of serum levels of Ang-2 and CC16 could be used for early diagnosis of ARDS, and the combined detection of both has a higher sensitivity than single detection.

Application of early sequential unclamping method in laparoscopic partial nephrectomy for patients with T1b renal tumor / 中华泌尿外科杂志

Objective To analyze the value of early sequential unclamping method in laparoscopic partial nephrectomy.Methods From April 2017 to October 2017,a total of 8 cases of renal tumor patients by early sequential unclamping method of laparoscopic partial nephrectomy (LPN) were reviewed,with 5 males and 3 females and average age of 56.4 years (43-70 years).Three cases of renal tumor were located on the left side,5 cases on the right side.The mean tumor diameter was 5.6 (4.6-6.4) cm.The preoperativeR.E.N.A.L.score was 8.8 (7-10),and the mean ASA score was 1.4 (1-2).Preoperative serum creatinine level was 89.5 (72.1-104.2) μmol/L,and the GFR level of the kidney with tumor before operation was 55.5 (40.4-62.3) ml/min.The early sequential unclamping method was used for retroperitoneal laparoscopic partial nephrectomy:according to the preoperative CTA results,the main branches and branches of the renal artery were routinely separated.Before the tumor resection,the branches of renal artery and the main renal artery were sequentially blocked.After removal of the tumor,the first layer of bare kidney wound blood vessels and collection system were sutured and repaired.Then released the main renal artery occlusion clamp,restored most of the blood supply to the kidney,but kept the tumor-specific segmental renal artery blocked.Continuous suture of the kidney created a rough combination of the renal wound.After second layers of suture completed,unclamped the segmental renal artery and sutured the renal wound again,made the third layers of suture intersecting with the second seam suture to strengthen the hemostatic effect.Results All the 8 patients were performed LPN with early sequential unclamping method successfully.The average operative time was 132.5 (90-180) min,the intraoperative blood loss was 142.5 (100-200) ml,the completely warm ischemia time was 15.5 (12.0-20.0) min,and no blood transfusion was performed intraoperatively and postoperatively.The operative margin was negative.The postoperative pathology showed that 7 cases were clear cell carcinoma and 1 cases of papillary cell carcinoma.Postoperative complications such as urinary leakage,incision infection and fever were not found.Drainage tube removal time was 3.5 (3-5) days and the time of postoperative hospitalization was 4.8 (4-6) days.At 1 months after operation,the serum creatinine level was 94.0 (83.6-101.2) μmol/L and the GFR level of one side kidney with tumor was 52.3 (43.2-59.6) ml/min.After 2-9 months of follow-up,there was no recurrence of the tumor.Conclusions Early sequential unclamping method could shorten the warm ischemia time and reduce the risk of bleeding during the operation.It also maintains a clear operative field,which could reduce the difficulty of laparoscopic partial nephrectomy and make a more accurate tumor resection in the complex renal tumor patients.

Differential expression profile of long non-coding RNA in the lipopolysaecharide-induced inflammation of monocyte-derived macrophages / 中华危重病急救医学

Objective To analyze the expression profile of long non-coding RNA (lncRNA) in the lipopolysaecharide (LPS)-induced inflammation of monocyte-derived macrophages.Methods Peripheral blood mononuclear cells were derived from healthy donor and induced into macrophages. The macrophages were divided into blank control group and LPS (1 mg/L) stimulated 12 hours group. Culture supernatants and cell pellets were harvested in each group, enzyme linked immunosorbent assay (ELISA) was used to assay the production changes of interleukins (IL-1β and IL-6), and tumor necrosis factor-α (TNF-α) in the supernatant. The technique of lncRNA microarray was used to test the lncRNA expression profile in LPS-induced inflammation of macrophages and control macrophages. The raw data of lncRNA were pretreated for normalization. Five lncRNA expressions were validated by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Furthermore, qRT-PCR was used to detect the expression of NR_028034 in macrophages after LPS-induced inflammation.Results ① The contents of IL-1β (ng/L:562.93±61.17 vs. 59.74±15.68), IL-6 (ng/L: 702.46±92.31 vs. 71.66±18.25) and TNF-α (ng/L: 794.50±63.89 vs. 85.12±22.07) in the LPS group were significantly higher than those in the blank control group (allP < 0.01). These results indicated that the inflammatory model of human macrophages was constructed successfully. ② Compared with blank control group, and 1479 lncRNA which have more than 2 folds variation and significant difference (P < 0.05) by statistical analysis was defined as lncRNA with differential expression. Among these lncRNA, LPS group showed 953 up- regulated and 526 down- regulated genes by 2 folds and 49 up- regulated and 35 down- regulated genes by 5 folds. ③ qRT-PCR results were generally consistent with the microarray data. ④ The expression of NR_028034 was increased by (4.41±0.65), (11.56±2.04), (18.58±1.36) folds compared with blank control group at 3, 6, 12 hours after LPS stimulation (allP < 0.01).Conclusions These data show a significantly altered lncRNA expression profile in the LPS-induced inflammation of monocyte-derived macrophages, suggesting that lncRNA may be involved in regulation of macrophages inflammatory response.

Application of 3D printing technique combined with intra-operative ultrasound for completely endophytic renal tumor in laparoscopic surgery / 第二军医大学学报

Objective To explore the feasibility of 3D printing technique combined with intra-operative ultrasound for locating completely endophytic renal tumor in lapamscopic nephron-sparing surgery.Methods Fifteen patients with completely endophytic renal tumor,who undervwent 3D printing technique combined with intra-operative ultrasound assisted localization of laparoscopic partial nephrectomy from Mar.2014 to Mar.2016,received CT image 3D reconstruction and 3D printing kidney model using Fommlab Form1 + 3D printer before operation.Among 15 patients aging (55.7±10.5) years,11 patients were male and 4 were female;the tumor diameter was (2.8±1.0) cm;and 3 cases were ventral and 12 were dorsal,all had solitary tumors.The clinical data,including intra-operative blood loss,warm ischemia time,post-operative pathology and surgical margins,and post-operative renal function,were statistically analyzed in this study.Resuits Laparoscopic partial nephrectomy was successfully carried out in all cases:the average operation time was (105.0± 20.6) min,the average warm ischemia time was (22.8 ± 3.5) min,and the mean intra-operative blood loss was (87.3±15.8) mL.No case received blood transfusion during or after operation,and the average post-operative hospital stay was (6.7 ± 1.0) days.No obvious complication occurred after operation.The surgical margins were all negative.Post-operative pathology confirmed that 13 patients were with clear cell renal cell carcinoma,and 2 with papillary renal cell carcinoma.Patients were followed up for (23.7± 11.8) months,and nocontinuous deterioration of renal function or tumor recurrence was found.Conclusion Pre-operative 3D printing technique for patients with completely endophytic renal tumor can help to determine the tumor location and adjacent relationship,reducing the risk of surgery by guiding operation scheme.Meanwhile,propaganda and education using 3Dprinting kidney model can improve patients ' cognition to surgery and simplify pre-operative conversation process.Furthermore,utilizing intra-operative ultrasound to optimize tumor resection scheme can reduce the damage to the renalvessels and collection system,maximizing the clinical benefit by ensuring negative margin and renal function reservation.

Effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages / 中华微生物学和免疫学杂志

Objective To investigate the effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages by analyzing the expression of lincRNA-cox2 in RAW264. 7 macrophages of M1 and M2 phenotypes. Methods Murine RAW264. 7 cells were induced by IFN-γand LPS to polarize to M1 phenotype, and were induced by IL-4 to polarize to M2 phenotype. The expression of lincRNA-cox2 in M1 and M2 macrophages were analyzed by real-time quantitative PCR ( RT-PCR) . We designed and synthesized siRNA oligo for lincRNA-cox2 and unrelated sequences. Then the siRNA oligo and NC oligo were transfected into RAW264. 7 cells by LipofectmineTM 2000. The transfected RAW264. 7 cells were induced by IFN-γand LPS or by IL-4 to polarize to M1 or M2 macrophages. Enzyme linked immunosorbent assay ( ELISA) was performed to measure the secretion of IL-10 and IL-12 induced in different conditions. The expression of in-ducible nitric oxide synthase ( iNOS ) , TNF-α, arginase 1 ( Arg-1 ) and found in inflammatory zone 1 (Fizz1) at mRNA level were detected by RT-PCR. The M1 macrophages were transfected with siRNAs to knock down the expression of lincRNA-cox2 for analyzing the biological effects of lincRNA-cox2 on the polar-ization of macrophages. Results The relative expression of lincRNA-cox2 in M1 macrophages was signifi-cantly higher than that in RAW264. 7 cells and M2 macrophages. Compared with the control group, the RAW264. 7 cells transfected with lincRNA-cox2-siRNA showed decreased secretion of IL-12 and inhibited expression of iNOS and TNF-αat mRNA level after IFN-γand LPS induction, but increased secretion of IL-10 and enhanced expression of Arg1 and Fizz1 at mRNA level after IL-4 induction. Transfecting the M1 mac-rophages with lincRNA-cox2-siRNA inhibited the secretion of IL-12, but promoted the secretion of IL-10. Conclusion This study indicated that lincRNA-cox2 was involved in the regulation of macrophage pheno-types by promoting the polarization to M1 macrophages and inhibiting the polarization to M2 macrophages.

Lentivirus-mediated RNA interference targeting TACO gene increases the intracellular killing of Mycobacterium tuberculosis by promoting the fusion of bacteria-containing phagosomes and lysosomes / 中华微生物学和免疫学杂志

Objective To construct a lentiviral vector-based short hairpin RNA (shRNA) targeting the gene encoding tryptophan-aspartate containing coat protein ( TACO) and to evaluate its inhibitory effect on the expression of TACO , and to further elucidate its effects on the phagocytosing and intracellular killing of My-cobacterium tuberculosis (M.tb) by macrophages and the possible mechanisms.Methods Three shRNA frag-ments targeting TACO gene and a scrambling control shRNA fragments were designed and cloned into the lentivi -ral vector pSicoR .The recombinant lentiviral vectors were identified by sequencing analysis and then packed in 293T cells.Real-time RT-PCR and Western blot assay were performed to evaluate the gene-silencing efficiency of the recombinant lentiviral vectors among RAW 264.7 cells transfected with the concentrated lentivirus .The most effective lentivirus was screened out to transfect the RAW 264.7 cells for 48 hours, followed by infection those cells with M.tb strains.The entry and intracellular survival of M .tb strains in RAW264.7 cells were de-termined by bacterial culture at indicated time points .The colocalization of M .tb and lysosomes was detected by immunofluorescence staining .The cyto-ID autophagy kit was used to detect the cellular autophagy and the auto-phagy-associated protein LC 3 was determined by Western blot assay .Results The recombinant lentiviral vec-tors were successfully constructed and confirmed by sequencing analysis .Decreased expression of TACO in RAW264 .7 cells was detected after transfecting the cells with the lentiviral vector-based shRNA vectors targeting TACO gene for 48 hours.The most effective lentivirus , LV-pSRT1, decreased the expression of TACO by 85.24%and 69.00%at the mRNA and protein levels, respectively.The bacterial loads in LV-pSRT1 trans-fected RAW264.7 cells were significantly decreased at the time point of 0 h after M.tb infection as compared with those in the control lentivirus treated cells (5.50×104 vs 8.1 ×104, P<0.05).Compared with the RAW264 .7 cells transfected with control lentivirus , the survival rate of intracellular M .tb strains in LV-pSRT1 transfected cells was significantly decreased at the time point of 48 h (134.54% vs 213.58%, P<0.05) and 72 h (148.18%vs 262.96%, P<0.05) considering the bacterial load at the time point of 0 h as the standard. The immunofluorescence staining demonstrated that the colocalization of M .tb strains with lysosomes was signifi-cantly enhanced in LV-pSRT1 transfected cells as compared with that in control lentivirus treated cells (75.67%vs 10.66%, P<0.05).Moreover, significantly enhanced autophagy and relative expression of LC 3Ⅱ protein were observed in RAW264.7 cells with TACO gene knockdown (16.20%vs 8.50%, P<0.05;0.51 vs 0.34, P<0.05).Conclusion The lentiviral vector-based shRNA targeting TACO gene could effectively knockdown the expression of TACO protein , decrease the entry and increase the intracellular killing of M .tb strains in mac-rophages.The enhanced intracellular killing of M .tb strains by macrophages was associated with the increased fusion of M.tb-containing phagosome and lysosome .

Clinical Efficacy of Integrative Medicine in Treatment of Nine HIV/AIDS-associated Non-infectious Skin Lesion Cases / 世界科学技术-中医药现代化

This study was aimed to observe the clinical efficacy of Highly Active Antiretroviral Therapy (HAART) combined withPing-Ai(PA) Mixture in the treatment of Human Immunodeficiency Virus / Acquired Immunodeficiency Syndrome (HIV/AIDS) patients with non-infectious skin lesions, in order to study the advantages of integrative medicine in AIDS prevention and treatment. A total of 9 cases, who met the inclusion criteria of AIDS-associated non-infectious skin lesion with the pattern ofqi-yin deficiency and internal obstruction of phlegm-stasis, were selected. The combination of HAART and number 1 and 4 prescription of PA Mixture were used in the treatment for 3 months. The observations were made on changes of clinical symptoms, body signs, Karovsky integral and CD4+ T cells of patients before and after treatment. Stata 12.0 was used in the statistical analysis. The results showed that after integrative therapy, the single integral of clinical symptoms and body signs, such as skin spot papula, subcutaneous nodules, skin itching, fatigue was obviously reduced compared to that of the pretreatment. The total score decreased than that of the pretreatment. The patients’ quality of life (QOL) significantly increased. The Karovsky integral increased compared with that of pretreatment. The CD4+ T cells values were higher than that of the pretreatment. It was concluded that the integrative therapy had a certain effect in the treatment of AIDS-associated non-infectious skin lesions, which can improve the clinical symptoms, body signs, obviously increase the patients’ QOL and immune function.

Advances on enzymes and enzyme inhibitors research based on microfluidic devices / 药学学报

With the continuous development in microfluidic fabrication technology, microfluidic analysis has evolved from a concept to one of research frontiers in last twenty years. The research of enzymes and enzyme inhibitors based on microfluidic devices has also made great progress. Microfluidic technology improved greatly the analytical performance of the research of enzymes and enzyme inhibitors by reducing the consumption of reagents, decreasing the analysis time, and developing automation. This review focuses on the development and classification of enzymes and enzyme inhibitors research based on microfluidic devices.