Blood molecular markers associated with COVID-19 immunopathology and multi-organ damage.

COVID-19 is characterized by dysregulated immune responses, metabolic dysfunction and adverse effects on the function of multiple organs. To understand host responses to COVID-19 pathophysiology, we combined transcriptomics, proteomics, and metabolomics to identify molecular markers in peripheral blood and plasma samples of 66 COVID-19-infected patients experiencing a range of disease severities and 17 healthy controls. A large number of expressed genes, proteins, metabolites, and extracellular RNAs (exRNAs) exhibit strong associations with various clinical parameters. Multiple sets of tissue-specific proteins and exRNAs varied significantly in both mild and severe patients suggesting a potential impact on tissue function. Chronic activation of neutrophils, IFN-I signaling, and a high level of inflammatory cytokines were observed in patients with severe disease progression. In contrast, COVID-19-infected patients experiencing milder disease symptoms showed robust T-cell responses. Finally, we identified genes, proteins, and exRNAs as potential biomarkers that might assist in predicting the prognosis of SARS-CoV-2 infection. These data refine our understanding of the pathophysiology and clinical progress of COVID-19.

Moderate-dose glucocorticoids as salvage therapy for severe pneumonia in renal transplant recipients: a single-center feasibility study.

This study aimed to assess the effectiveness and safety of moderate-dose glucocorticoids (GCs) with mechanical ventilation as salvage therapy for renal transplant recipients with severe pneumonia, which was non-responsive to conventional treatment. A retrospective study was conducted involving renal transplant recipients diagnosed with severe pneumonia and did not respond to conventional treatment. All immunosuppressants were then completely withdrawn, and the patients were initially administered with methylprednisolone at doses of 2.0-2.5 mg/kg/day once every 12 h. This dosage was continued until oxygenation improved, and the treatment was gradually tapered (by 20 mg every 2-3 days) to the previous maintenance dosage. Ten patients were recruited from year 2008 to 2012. Two patients who underwent emergency endotracheal intubation were intubated on days 3 and 8, respectively, another one died from recurrent pneumothorax. The mean PaO2/FiO2 of the nine survivors was significantly increased by the increasing treatment duration; whereas the lung injury scores (LIS) and the sequential organ failure assessment (SOFA) score were both significantly decreased. The use of moderate-dose GCs may play a role as salvage therapy for renal transplant recipients with severe pneumonia. However, further study with larger trials to is needed.

Combination of caspofungin and low-dose trimethoprim/sulfamethoxazole for the treatment of severe Pneumocystis jirovecii pneumonia in renal transplant recipients.

Pneumocystis jirovecii pneumonia (PJP) is a severe and life-threatening complication in immunocompromised patients. Trimethoprim/sulfamethoxazole (TMP-SMZ) is well known for its effectiveness as prophylaxis of PJP. However, the use of TMP-SMZ is associated with various adverse effects that may not be tolerated by critically ill patients. Caspofungin is recommended for invasive fungal infections, but the treatment of PJP after solid organ transplantation (SOT) is an off-label use of this drug. In this study, three cases of severe PJP in renal transplant recipients treated with a combination of caspofungin and low-dose TMP-SMZ were presented. Initial findings indicated that the combined treatment may be beneficial for the treatment of PJP and decrease the incidence of TMP-SMZ-related adverse effects.

[Expression of inflammation cytokines and network analysis in acute rejection of renal transplantation].

OBJECTIVE: To explore the changes of inflammation cytokines during acute renal transplantation rejection and decipher the functions of their protein-protein interaction network. METHODS: Serum samples were collected from renal transplantation patients with stable renal function or acute rejection (n = 6 each) to measure the expression level of 40 inflammatory factors by APIX protein array. The differentially expressed proteins were selected and their protein-protein interaction networks constructed. And biologic processes were analyzed by the online tools of String and Network Ontology Analysis. RESULTS: There were 8 differentially expressed cytokines in the AR group versus the stable group (M (Q(1)-Q(3)), CCL24: 700 (255 - 1157) vs 330 (100 - 610) ng/L, ICAM-1: 58 737 (8018 - 105 395) vs 22 660 (137 - 68 914) ng/L, IL-10: 120 (20 - 517) vs 298 (81 - 11 609) ng/L, IL-6sR: 11 328 (3357 - 21 251) vs 7665 (370 - 12 455) ng/L, CCL3: 1712(7002 - 32 634) vs 283 (54 - 1915) ng/L, CCL4: 554 (28 - 2355) vs 283(104 - 1915) ng/L, TIMP-1: 15 560 (13 343 - 42 481) vs 11 271 (1207 - 18 228) ng/L, CCL5: 44 547 (38 252 - 78 631) vs 27 765 (12 073 - 46 627) ng/L, all P < 0.05). The analyses of protein-protein association network showed that these proteins were correlated and involved in such biological processes as taxis, chemotaxis, inflammatory reactions, wound responses and leukocytic migration. CONCLUSIONS: Comparing the inter-group differences of inflammatory cytokines and further developing and analyzing the protein-protein interaction network may help us to explore the mechanisms of acute renal transplantation rejection. And the differential cytokines can be used as candidate diagnostic biomarkers and intervention targets.

Radiotherapy may improve overall survival of patients with T3/T4 transitional cell carcinoma of the renal pelvis or ureter and delay bladder tumour relapse.

BACKGROUND: Since transitional cell carcinoma (TCC) of the upper urinary tract is a relatively uncommon malignancy, the role of adjuvant radiotherapy is unknown. METHODS: We treated 133 patients with TCC of the renal pelvis or ureter at our institution between 1998 and 2008. The 67 patients who received external beam radiotherapy (EBRT) following surgery were assigned to the radiation group (RT). The clinical target volume included the renal fossa, the course of the ureter to the entire bladder, and the paracaval and para-aortic lymph nodes, which were at risk of harbouring metastatic disease in 53 patients. The tumour bed or residual tumour was targeted in 14 patients. The median radiation dose administered was 50 Gy. The 66 patients who received intravesical chemotherapy were assigned to the non-radiation group (non-RT). RESULTS: The overall survival rates for the RT and non-RT groups were not significantly different (p = 0.198). However, there was a significant difference between the survival rates for these groups based on patients with T3/T4 stage cancer. A significant difference was observed in the bladder tumour relapse rate between the irradiated and non-irradiated bladder groups (p = 0.004). Multivariate analysis indicated that improved overall survival was associated with age < 60 years, T1 or T2 stage, absence of synchronous LN metastases, and EBRT. Acute gastrointestinal and bladder reactions were the most common symptoms, but mild non-severe (> grade 3) hematologic symptoms also occurred. CONCLUSION: EBRT may improve overall survival for patients with T3/T4 cancer of the renal pelvis or ureter and delay bladder tumour recurrence in all patients.

[Analysis of microchimerism in living renal transplantation between mothers and children relationship].

OBJECTIVE: To compare the microchimerismic and rejection rates in living donor kidney transplant recipients in mother and child relations and other relations. METHODS: This retrospective single-center study enrolled 130 recipients to receive allografts from living related donors from 2004 to 2008 at our hospital. They were followed up for 1 - 5 years. The demographic data of the study population were analyzed by basic statistical methods. A total of 43 recipient blood samples were collected for the detection of microchimerism by the assays of short tandem repeat (STR) and sex-determining region-y gene (SRY) polymerase chain reaction (PCR). RESULTS: The 1-year patient/graft survival rates were 93.8% and 92.3% respectively. And there was no significant differences between mother and child group and other relative group. Forty-six biopsy samples were collected from 46 recipients. Twenty-six (20.0%) cases had the occurrences of acute rejection episodes in different Banff degrees as proven by biopsy. 53.8% (14/26) cases were mother and child renal transplantation, higher than other relative (46.2%, 12/26). The mother donor kidney transplant recipients had about a twice higher rejection rate (30.4% vs 14.3%, P = 0.028) and a twice higher microchimerismic rate (25.0% vs 14.8%) than other relative. CONCLUSION: Compared with other relations, the mother donor kidney recipients tend to have higher rates of microchimerism and acute rejection. And the special immune effect in mothers and children renal transplantation may influence its outcomes.

[Preliminary exploration in proteomic-based mechanistic analysis of patients with acute renal allograft rejection].

OBJECTIVE: To explore the application of proteomics in the mechanistic analysis of acute rejection (AR). METHODS: Quantified proteomics with isobaric tags for relative and absolute quantitation (iTRAQ) labeling was utilized to identify the protein profiling between the transplantation patients with (n = 5) or without AR (n = 8) from 2008 to 2010. RESULTS: Among the 179 identified proteins, 66 proteins in AR patients had at least a 2-fold change as compared with those without AR. The results demonstrated the dominant processes and responses associated with inflammation and complement activation. It was consistent with the underlying immune rejection associated with AR. Moreover, the results also indicated that high-coagulation state existed in AR patients. A number of transcription factors were identified in AR patients, including nuclear factor-κB, signal transducer and activator of transcription 1, signal transducer and activator of transcription 3. The analysis of transcription regulation networks suggested that the cross-talks among these key transcription factors might play an important role in the acute response and activation of coagulation system. CONCLUSION: The application of proteomics provides a new strategy of mechanistic analysis in AR.

[Long-term effect on radiofrequency heat-coagulation endometrial ablation in the treatment of anovulatory dysfunctional uterine bleeding].

OBJECTIVE: To investigate long-term effect on radiofrequency heat-coagulation (RF) endometrial ablation in treatment of anovulatory dysfunctional uterine bleeding (DUB). METHODS: From Jul. 2001 to Nov. 2009, 1196 patients with DUB who were failed by medical treatment (including 127 patients with dysmenorrheal) were enrolled into this study in Jinan Millitary General Hospital. Those patients were divided into two groups according to age: 427 patients at age of or more than 45 years (average age 48 years) in Group A who were treated by RF procedure for amenorrhea;769 patients at age of less than 45 years old (average 37 years) in group B were treated by RF for controlling excessive menstrual bleeding. All the patients had the results of menstrual score (pictorial blood loss assessment chart, PBAC), hemoglobin (Hb), endometrial curettage pathology and hysteroscopy examination immediately after RF procedure; Some patients still had another endometrial curettage pathology and clinical results in 6 months after RF. The mean follow-up time was 72 months (range: 6 to 100 months). The evaluation criterion for RF treatment was to use optimal and significant effect measurements. For group A, the optimal treatment effect (cure) was defined as bleeding cessation and achieving amenorrhea that continued for more than 12 months after treatment. For group B, the optimal treatment effect(cure) was also defined as bleeding cessation and resuming normal menstruation which continued for more than 12 months after treatment. Significant treatment effect was defined as irregular, minor bleeding, but PBAC score less than 100 within 12 months. If patient symptoms and PBAC scores did not change compared with those before treatment, the treatment was defined as failure. For dysmenorrhea, the optimal treatment effect was disappearance for more than 12 months, the significant treatment effect was remission, and treatment failure was not changed from the pre-treatment baseline. The effective rate was the sum of that of the optimal and significant effect. One hundred and twenty-five patients with DUB treated by agents at the same time were chosen as control group. RESULTS: (1) The recent and long-term effective rates for bleeding cessation by RF:the total recent effective rates within 1 months were 94.82% (1134/1196), including 96.5% (412/427) in group A and 93.9% (722/769) in group B. The total curative rates for dysmenorrheal were 82.7% (105/127), including 86.4% (38/44) in group A and 80.7% (67/83) in group B. Pathology examination after hysteroscopy immediately after RF showed a completely and whole destroyed endometrium in group A, and a little rested endometrium in group B. The long-term effect rates for bleeding cessation by RF after 12, 24 and 36 months were 92.55% (969/1047), 93.9% (866/922) and 93.7% (609/650), respectively. PBAC and Hb in group A and group B within 12, 24, 36 and more than 36 months were improved significantly (P < 0.05). (2) COMPLICATIONS: the major complication was irregular minor bleeding in 1 to 2 months after treatment, the rate was 8.03% (96/1196). The second one was menorrhea in 3 months after RF, the rate was 5.18% (62/1196). This condition was corrected by the second RF. No hysterectomy was performed on those patients. CONCLUSION: RF is the safe, efficient and minimal invasive procedure in treatment for DUB. The mechanism of keeping long-term curative effect and preventing recurrence is due to endometrium inactivation and fibrosis by thermocoagulation.

Identifying the Risk of SARS-CoV-2 Infection and Environmental Monitoring in Airborne Infectious Isolation Rooms (AIIRs).

Healthcare workers (HCWs) are at high risk of occupational exposure to the new pandemic human coronavirus, SARS-CoV-2, and are a source of nosocomial transmission in airborne infectious isolation rooms (AIIRs). Here, we performed comprehensive environmental contamination surveillance to evaluate the risk of viral transmission in AIIRs with 115 rooms in three buildings at the Shanghai Public Health Clinical Center, Shanghai, during the treatment of 334 patients infected with SARS-CoV-2. The results showed that the risk of airborne transmission of SARS-CoV-2 in AIIRs was low (1.62%, 25/1544) due to the directional airflow and strong environmental hygiene procedures. However, we detected viral RNA on the surface of foot-operated openers and bathroom sinks in AIIRs (viral load: 55.00-3154.50 copies/mL). This might be a source of contamination to connecting corridors and object surfaces through the footwear and gloves used by HCWs. The risk of infection was eliminated by the use of disposable footwear covers and the application of more effective environmental and personal hygiene measures. With the help of effective infection control procedures, none of 290 HCWs was infected when working in the AIIRs at this hospital. This study has provided information pertinent for infection control in AIIRs during the treatment of COVID-19 patients.

[The clinical characteristic of adrenal metastatic tumor].

OBJECTIVE: To analyze the clinical features of adrenal metastasis. METHODS: From January 1993 to December 2004, 103 cases of adrenal metastasis were reviewed. RESULTS: Lung and hepatocellular carcinoma were the most common primary tumor of adrenal metastatic tumor, which about 36.9% (38/103) and 42.7% (44/103) of all cases, followed by renal carcinoma 6.8% (7/103), colorectal carcinoma 4.9% (5/103), stomach carcinoma 3.9% (4/103), breast cancer 1.9% (2/103), unknown primary tumor 2.9% (3/103). Most of these were low differentiation. The mean diameter of adrenal metastasis was 3.9 cm. The mean interval from detection of primary tumor to adrenal metastasis was 9.5 months. And 79.6% (82/103) were detected as a part of multiorgan metastasis. Only 5 cases (4.9%) were presented with pain in the back. There was little characterization of ultrasonography, CT and MRI, color-Doppler and selective arterial imaging showed little blood supply. All of patients were treated with synthetic methods, 16 cases (15.5%) who had undergone adrenalectomy for metastasis disease had a improved survival compared with those non-adrenalectomy. CONCLUSIONS: There is no particular presentation of clinic and imaging, diagnosis depending on history, follow-up and the pathological presentation of primary tumor. There are no standard treatment guidelines for this group of patients. When the primary tumor could be resected or be well controlled, and there is no other evidence of metastasis, adrenalectomy is recommended. Transarterial chemoembolization (TACE) could not actually be performed.

RETRACTED: Twist2 promotes kidney cancer cell proliferation and invasion via regulating ITGA6 and CD44 expression in the ECM-Receptor-Interaction pathway.

Twist2 is a member of the basic helix-loop-helix (bHLH) family and plays a critical role in tumorigenesis. Growing evidence proves that Twist2 involves in tumor progression; however, the role of Twist2 in human kidney cancer and its underlying mechanisms remain unclear. RT-PCR and Western blot analysis were used to detect the expression of Twist2 in kidney cancer cells and tissues. Cell proliferation, cell cycle, apoptosis, migration and invasion assay was measured by the Cell Count Kit-8 (CCK8), flow cytometry, wound healing and transwell analysis, respectively. Gene set enrichment analysis (GSEA) was used to identify correlation of Twist2 with ECM-Receptor-Interaction pathway. In this report, we show that Twist2 up-regulated in human kidney cancer tissues compared with normal kidney tissues. Twist2 promotes cell proliferation, inhibits cell apoptosis, augments cell migration and invasion in human kidney cancer-derived cell in vitro, and promotes tumor growth in vivo. Moreover, we found that knockdown of Twist2 decreased the levels of ITGA6 and CD44 which contribute to cell migration and invasion correlated with ECM-Receptor-Interaction pathway. This result indicates Twist2 may promote migration and invasion of kidney cancer cells by regulating ITGA6 and CD44 expression. Therefore, our data demonstrated that Twist2 involves in kidney cancer progression. The identification of the role Twist2 on the migration and invasion of kidney cancer provides a potential appropriate treatment after radical nephrectomy to get a better prognosis that reducing recurrence.

Twist2 promotes kidney cancer cell proliferation and invasion by regulating ITGA6 and CD44 expression in the ECM-receptor interaction pathway.

Twist2 is a member of the basic helix-loop-helix (bHLH) family and plays a critical role in tumorigenesis. Growing evidence has proven that Twist2 is involved in tumor progression; however, the role of Twist2 in human kidney cancer and its underlying mechanisms remain unclear. Real-time polymerase chain reaction and Western blot analysis were used to detect the expression of Twist2 in kidney cancer cells and tissues. Cell proliferation, cell cycle, apoptosis, migration, and invasion assay were analyzed using the Cell Count Kit-8, flow cytometry, wound healing, and Transwell analysis, respectively. In this study, we showed that Twist2 was upregulated in human kidney cancer tissues compared with normal kidney tissues. Twist2 promoted cell proliferation, inhibited cell apoptosis, and augmented cell migration and invasion in human kidney-cancer-derived cells in vitro. Twist2 also promoted tumor growth in vivo. Moreover, we found that the knockdown of Twist2 decreased the levels of ITGA6 and CD44 expression. This result indicates that Twist2 may promote migration and invasion of kidney cancer cells by regulating ITGA6 and CD44 expression. Therefore, our data demonstrated that Twist2 is involved in kidney cancer progression. The identification of the role of Twist2 in the migration and invasion of kidney cancer provides a potential appropriate treatment for human kidney cancer.

[Effect of astrgaloside IV on the long-term consequences of renal ischemia-reperfusion injury in rat].

OBJECTIVE: To observe the effect of astrgaloside IV (Astr) on the long-term consequences of renal ischemia-reperfusion injury (IRI) in rat. METHODS: Fifty-four male Sprague-Dawley rats were randomized into 3 equal groups: IRI group, Astr group, and sham operation group. All rats underwent right nephrectomy and isolation of the left renal artery. The left renal arteries of the IRI group and Astr group were gripped by vascular clamp for 60 minutes and that of the sham operation group was only isolated without gripping. Two milliliters of Astr solution (0.1 g/L) was perfused into the stomach of the rats in the Astr group three days before and after the operation respectively. The rats in the IRI and sham operation groups were perfused with normal saline of the same volume. Four, twelve, and twenty-four weeks after the operation 24-hour urine specimens of the rats were collected to detect the urine protein. At each time point 6 rats from each group were anesthetized and blood was collected from the abdominal aorta to measure the level of serum creatinine (Cr), their left kidneys were taken out to undergo pathological examination and extraction of mRNA. Histochemistry was used to detect the expression of tumor growth factor (TGF)-beta1 protein in the renal tissues. RT-PCR was used to detect the expression of TGF-beta1 mRNA. Collagen staining and immunohistochemistry were used to measure the proportion of collagen positive material to the total area. RESULTS: The level of urine protein was increased progressively, those 12 and 24 weeks after the operation in the IRI group were significantly higher than those in the Astr and sham operation groups (all P < 0.05). The serum Cr 4 weeks after the operation was 36 micromol/L +/- 4 micromol/L, significantly higher than those in the Astr and sham operation groups (31 micromol/L +/- 8 micromol/L and 31 micromol/L +/- 5 micromol/L), and the serum Cr levels 4 weeks 12 and 24 weeks after the operation in the IRI group remained significantly higher than those in the Astr and sham operation groups (all P < 0.05). Collagen staining showed that the glomerular basement membrane, tunica adventitia vasorum, and adventitia of renal tubule were remarkably redder in the IRI than in the Astr and sham operation groups. The expression of TGF-beta1 protein was progressively increased since 12 weeks after the operation in the IRI group, significantly stronger in the Astr and sham operation groups. The expression of TGF-beta1 mRNA was progressively increased since 12 weeks after the operation in the IRI and Astr groups, significantly stronger than that in the sham operation group (P < 0.05). However, the expression of TGF-beta1 mRNA 24 weeks after the operation was significantly stronger in the IRI group than in the Astr group (P < 0.05). CONCLUSION: After renal IRI the probability of development of renal fibrosis increases. Astrgaloside IV markedly ameliorates renal injury by downregulating the TGF-beta1 expression.

Antithymocyte globulin-induced acute respiratory distress syndrome after renal transplantation: a case report.

Antithymocyte globulin (ATG) has long been used for immune-induction and anti-rejection treatments for solid organ transplantations. To date, few cases of ATG-induced acute respiratory distress syndrome (ARDS) have been published. Here, we present a case of ARDS caused by a single low-dose of ATG in a renal transplant recipient and the subsequent treatments administered. Although the patient suffered from ARDS and delayed graft function, he was successfully treated. We emphasize that the presence of such complications should be considered when unexplained respiratory distress occurs. Early use of corticosteroids, adjustment of immunosuppressive regimens, and conservative fluid management, as well as empiric antimicrobial therapies, may be effective strategies for the treatment of ARDS caused by ATG.

Prediction of acute cellular renal allograft rejection by urinary metabolomics using MALDI-FTMS.

The present study investigated small molecule analysis of urinary samples as a noninvasive method to detect acute cellular renal allograft rejection. Matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS) was used to analyze 15 urinary samples from transplant patients with different grades of biopsy showing improved clinical acute cellular rejection (ACR) and 24 urinary samples from 8 transplant patients without evidence of rejection. Seven small molecules demonstrated highly successful diagnostic performance (m/z): 278.1 (t = 3.398, p = 0.004), 293.0 (t = 2.169, p = 0.048), 294.1 (t = 2.154, p = 0.05), 382.2 (t = 2.961, p = 0.010), 383.3 (t = 2.270, p = 0.040), 402.2 (t = 2.994, p = 0.010), 424.0 (t = 2.644, p = 0.019). Kidney transplant patients with ACR could be distinguished from those without ACR using four individual small molecules with a specificity of 100%. In conclusion, the combination of MALDI-FTMS technology with a clear definition of patient groups can detect urine small molecule associated with ACR.