Next-generation sequencing technology for the diagnosis of Pneumocystis pneumonia in an immunocompetent female: A case report.

BACKGROUND: Pneumocystis pneumonia (PCP) is a serious fungal infection usually seen in patients with human immunodeficiency virus, and it is more frequently found and has a high fatality rate in immunocompromised people. Surprisingly, it rarely occurs in immunocompetent patients. However, the clinical diagnosis of this pathogen is made more difficult by the difficulty of obtaining accurate microbiological evidence with routine tests. This case reports a PCP patient with normal immune function who was diagnosed through next-generation sequencing (NGS). CASE SUMMARY: A 23-year-old female who had no special disease in the past was admitted to the hospital with a persistent fever and cough. Based on the initial examination results, the patient was diagnosed with bipulmonary pneumonia, and empirical broad-spectrum antibiotic therapy was administered. However, due to the undetermined etiology, the patient's condition continued to worsen. She was transferred to the intensive care unit because of acute respiratory failure. After the diagnosis of Pneumocystis jirovecii infection through NGS in bronchoalveolar lavage fluid and treatment with trimethoprim/sulfamethoxazole and caspofungin, the patient gradually recovered and had a good prognosis. CONCLUSION: This case emphasizes that, for patients with normal immune function the possibility of PCP infection, although rare, cannot be ignored. NGS plays an important role in the diagnosis of refractory interstitial pneumonia and acute respiratory failure.

[Clinical Analysis of Elderly Patients with AML/High-Risk MDS].

OBJECTIVE: To analyze the clinical features of acute myeloid leukemia (AML)/high-risk myelodysplastic syndrome (MDS) patients aged over 60 years old. METHODS: The clinical data of 61 elderly newly diagnosed patients with AML and high-risk MDS who submitted to the Department of Hematology/Oncology of the First Affiliated Hospital of Tsinghua University from January 2009 to April 16, 2021 were retrospectively analyzed. These patients were divided into chemotherapy group (45 cases) and supportive treatment group (16 cases). The overall survival (OS) was analyzed by Kaplan-Meier method, and the prognostic factors of survival were analyzed by multivariate Cox regression. RESULTS: After 2 cycles of induction chemotherapy, the complete remission (CR) rate was 37.8% (17/45), and overall response rate was 62.2% (28/45) in the chemotherapy group. The median OS in the chemotherapy group and supportive treatment group was 11.3 (0.07-43) and 1.6 (0.33-7.72) months, respectively (P<0.001). The median OS in patients who reached CR or did not reach after 1 cycle of induction chemotherapy was 19.8 (10-30.63) and 8.17 (0.07-43) months, respectively (P<0.05), while after 2 cycles was 22.7 (4.2-43) and 7.26 (0.07-26) months, respectively (P<0.001). Univariate analysis showed that age > 80 years old, CCI score > 2, PS score > 2 and supportive treatment were the adverse prognostic factors for OS. Further multivariate analysis suggested that chemotherapy was the only independent prognostic factor for OS (HR=0.140, 95%CI: 0.048-0.409, P<0.001). In the chemotherapy group, univariate analysis showed that CCI score > 2 and failure to reach CR after induction chemotherapy were poor prognostic factors. Multivariate analysis showed that CCI score > 2 (HR=0.139, 95%CI: 0.050-0.384, P<0.001) and failure to achieve CR after induction chemotherapy (HR=0.103, 95%CI: 0.041-0.259, P<0.001) were the adverse prognostic factors for OS. The patients were tolerant to side-effect of chemotherapy. CONCLUSION: Appropriate chemotherapy can prolong the survival of elderly patients with AML and high-risk MDS.

[Survival Analysis of 148 Patients with Primary Diffuse Large B Cell Lymphoma].

OBJECTIVE: To investigate the clinical outcome of the patients with primary diffuse large B-cell lymphoma（DLBCL）. METHODS: Clinical data of 148 patients with DLBCL in our hospital and cancer hospital from March 2006 to April 2016 were retrospectively analyzed. Kaplan-Meier analysis was used to estimate progression-free survival（PFS）and overall survival（OS）. RESULTS: 5-year OS and PFS rates were 85% and 69%，respectively. The survival analysis showed that 5-year OS rate of R-CHOP group was significantly higher than that of CHOP alone group（89% vs. 70%，P<0.05）. In univariate analysis，several clinical factors，such as older age（>60 years），poor ECOG score（≥2），advanced stage（Ⅲ-Ⅳ），higher IPI score（≥3），CHOP alone and absence of radiotherapy related with poor survival rate. Furthermore，multivariate analysis showed that age>60 year significantly related with the worse OS. CONCLUSION: Age >60 year is an important independent prognostic factor to predict worse OS or PFS.

[Effects of Lidocaine on the Rho/ROCK Signal Transduction of the Posterior Basilar Artery in Rabbit SAH].

OBJECTIVE: To observe the expression of Rho/Rho-associated kinase (ROCK) signaling pathway in the basilar artery and the effect of lidocaine on this signaling pathway after subarachnoid hemorrhage (SAH) in rabbits. METHODS: 24 New Zealand white rabbits were randomly divided into sham operation (sham) group, SAH group, and occipital cisterna lidocaine (CD) group. There were 8 rabbits in each group. Intracisternal injection of non-anticoagulant autologous arterial blood (1 mL/kg) were applied to SAH group and CD group animals to establish SAH model, sham normal saline group was injected with 37 ℃ physiological saline (1 mL/kg); after 30 min, CD group was injected with 0.3 mL 2% lidocaine cisterna, SAH group and sham group were injected with saline. After 72 h, food intake and neurologic function damage were measured. The expressions of Rho associated kinase 2 (ROCK2) and myosin light chain (MLC) and calmodulin (CaM) protein in the basilar artery were measured by Western blot. The ROCK2 and MLC and CaM gene expressions were measured by using real-time quantitative PCR. RESULTS: Compared with sham group, reduced food intake, various degrees of neurological impairment, increased ROCK2 mRNA and protein expressions in basal artery, and decreased MLC and CaM expressions were observed in SAH group and CD group (P<0.05). Compared with the SAH group, there was no statistically significant difference in diet intake and neurological damage in the CD group (P>0.05); the mRNA and protein expressions of ROCK2 in the basilar artery decreased, and the expressions of MLC and CaM increased (P<0.05). CONCLUSION: Intracisternal injection of lidocaine may inhibit the transmission of Rho/ROCK signal in the basilar artery and reduce the basilar artery smooth muscle contraction after SAH.

[Clinical Analysis of Priming Regimen for Treatment of Acute Myeloid Leukemia (Non-APL) Patients Aged over 70 Years].

OBJECTIVE: To explore the therapeutic efficacy for patients aged over 70 years with acute myeloid leukemia(non-APL). METHODS: Clinical data of 19 acute myeloid leukemia patients aged over 70 years admitted in our hospital from March 2006 to April 2016 years were analyzed retrospectively. Among them 15 patients received priming regimen and 4 patients received best supportive treatment. The overall survival time between patients with priming regimen and patients with best supportine treatment was compared by Kaplan-Meier analysis. The prognostic factors were evaluated by using multivariate analysis. Out of the 19 patients, 9 were males and 10 were females, with median age of 75 years (70-84). RESULTS: After 2 cycles of induction chemotherapy, the complete response rate was (7/15)46.6% and overall response rate was (11/15) 73%. Significant longer median survival time was observed in priming regimen group, compared with that in best supportive treatment group (11 months vs 2 months)(χ2=17.077, P<0.001). The patients were well tolerant to side-effect of chemotherapy. Multivariate analysis showed that only receiving induction chemotherapy or not was the independent prognostic factor of the survival time(P<0.05), while the sex, age, ECOG index, CD34, HLA-DR, WBC count and ratio of blast cells in bone marrow were factor affecting the prognosis. CONCLUSION: The priming regimen may be practicable and effective for the treatment of patients with acute myeloid leukemia aged over 70 years (non-APL) and prolong the survival time of these patients.

[Long Term Curative Effects and Prognostic Analysis of Stem Cell Transplantation for Treating Patients with Peripheral T Cell Lymphoma].

OBJECTIVE: To explore the efficacy and prognostic factors of hematopoietic stem cell transplantation in the treatment of patients with peripheral T cell lymphoma (PTCL). METHODS: The clinical records of 53 patients with PTCL were collected and analyzed retrospectively. The prognostic value of prognostic index PIT were evaluated. RESULTS: Among 53 cases of PTCL, 31 patients underwent hematopoietic autologous stem cell transplantation (auto-HSCT), 22 patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT), their median follow-up time was 62 (1-174) months, from them 1 patients was not engrafted in each group, 4 patients underwent allo-HSCT died within 1 month after transplantation. The 3-year overall survival rates in auto- and allo-HSCT groups were 42.3% (95% CI 24.1-60.5%) and 42.9% (95% CI 21.7-64.1%) respectively, 11 and 3 patients died of relapse after auto-HSCT and allo-HSCT, respectively, the treatment-related mortality (TRM) was 22.6% (95% CI 6.1%-39.0%) and 48.7% (95% CI 26.8%-70.7%), respectively. Univariate analysis showed that bone marrow involvement at diagnosis was associated with poor prognosis. In allo-HSCT group, PIT showed statistically significant difference in both OS and PFS between the patients without or with one risk factor and the patients with 3-4 risk factors. CONCLUSION: The relapse rate of relapse/refractory patients is high after auto-HSCT. The TRM is high after allo-HSCT, but long term survival is promising. The prognostic index for PTCL can reliably assess the prognosis of PTCL.

[Prevention Effect of Thymosin-alpha1 Aganist Early Ventilator-associated Pneumonia in Patients with Mechanical Ventilation].

OBJECTIVE: To investigate the preventive effects of thymosin-alpha1 against early ventilator-associated pneumonia (VAP) in the patients with mechanical ventilation. METHODS: Fifty two patients with expectancy of mechanical ventilation over 48 h were divided into routine therapy group (n=26) and thymosin therapy group (n= 26) in random. The patients in routine therapy group were given intensive care unit (ICU) conventional treatment, and the patients in thymosin therapy group were given thymosin treatment additionally (1.6 mg subcutaneous injection, qd X 7 d). The incidence and occurrence time of VAP were observed, and the time of mechanical ventilation and ICU stay were recorded. The levels of CD3+, CD4+, CD4+ /CD8+ T lymphocyte, CD14+ mononuclear cell human leukocyte antigens-DR (CD14+ HLA-DR) and procalcitonin (PCT) were detected before mechanical ventilation and at the 3d and 7th d after mechanical ventilation. RESULTS: The base line including the level of immunologic function had no difference between the two groups (P>0.05). The incidence of VAP in thymosin therapy group was lower than that in routine therapy group, but it was not significant difference (P>0.05). The durations of machine ventilation and ICU stay in thymosin therapy group were shorter than those in routine therapy group (P<0.05). The occurrence time of VAP in thymosin therapy group was significantly later than that in routine therapy group (P<0.05). At the 3rd and 7th d after mechanical ventilation, thymosin therapy group achived higher levels of CD3+, CD4+, CD4+ /CD8+ T lymphocyte and CD14+ HLA-DR than routine therapy group did (P<0.05). CONCLUSION: Thymosinal may be able to improve immunologic function and prevent the incidence of early VAP in the patients with mechanical ventilation.

[The effect of Xuebijing injection on immune regulation in SIRS patients].

OBJECTIVE: To investgate the effect of Xuebijing Injection on immune regulation in patients with systemic inflammatory response syndrome (SIRS). METHODS: 56 SIRS patients admitted to the ICU of Guizhou Provincial Hospital from January 2013 to December 2013 were included in this study. The patients were randomly divided into a control (C) and a treatment (T) group. Patients in C group received routine treatment; while patients in T group received additional Xuebijing Injection 50 mL Bid. The following indicators were recorded and compared between the two groups before and 4 and 7 days after treatments: CD4+, CD8+, CD4+/CD8+, monocytes CD14+/human leukocyte antigen-DR (HLA-DR), score of acute physiology and chronic health evaluation II (APACHE II), heart rate, white blood cells, body temperature, respiration rate, and MODS (7 d after treatment only). RESULTS: No differences were found between the two groups before treatments (P > 0.05). T Group had higher levels of CD4+, CD8, CD4+/CD8+ and monocytes CD14+/HLA-DR than C group at 4 and 7 d after treatments (P < 0.05). T group also had higher levels of improvement in vital indicators compared with C group (P < 0.05). No significant difference in incidence of multiple organ dysfunction syndrome (MODS) was found between the two groups (P > 0.05). CONCLUSION: Xuebijing Injection can regulate immune function of patients with SIRS.

Multifunctional Co(0.85)Se-Fe(3)O(4) nanocomposites: controlled synthesis and their enhanced performances for efficient hydrogenation of p-nitrophenol and adsorbents.

A new kind of multifunctional Co0.85 Se-Fe3 O4 nanocomposites is synthesized by loading Fe3 O4 nanoparticles (NPs) with a size of about 5 nm on the surface of Co0.85 Se nanosheets under hydrothermal conditions without using any surfactant or structure-directing agents. The Co0.85 Se-Fe3 O4 nanocomposite exhibits remarkable catalytic performance for hydrogenation of p-nitrophenol (4-NP) at room temperature and good adsorption behavior for methylene blue trihydrate in water. This nanocomposite also shows a high specific surface area and magnetic separation capability for recyclable utilization. The enhanced performances both in catalysis and adsorption are better than either individual component of Co0.85 Se nanosheets or Fe3 O4 nanoparticles, demonstrating the possibility for designing new multifunctional nanocomposites with improved performances for catalysis, adsorbents, and other applications.

[Clinical observation of decitabine-treating patients with myelodysplastic syndrome and acute myeloid leukemia].

This study was purposed to investigate the clinical efficiencies and adverse reactions of treating the myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) by using decitabine. The clinical data of 12 MDS and AML patients treated with decitabine were analyzed retrospectively. Among 12 patients there were 1 case of MDS-RA, 2 cases of MDS-RAEB-I, 3 cases of MDS-RAEB-II, 2 cases of AML-M4, 2 cases of AML-M5, 1 case of AML-M6 and 1 case of AML-M0. In decitabine chemotherapy program for 5 days (n = 8), decitabine 20 mg/(m(2)·d) × 5 days was applied, 4 weeks for 1 cycle; in program for 3 days (n = 2), decitabine 15 mg/m(2), once 8 h for 3 days, 6 weeks for 1 cycle; another program (n = 2), decitabine 20 mg/(m(2)·d) every other day for 5 times. For 1 patient achieved complete remission (CR) after treatment with decitabine, ID4 gene methylated level was detected by MS-PCR and ML-PCR before and after treatment. The results showed that 2 cases achieved CR, 1 case partial remission, 5 cases stable disease, 1 case progress of disease and 3 cases died. Disease control rate was 66.67% (8/12), the effective rate 25% (3/12). The average survival time was (11.5 ± 2.1) months. 1-year OS rate was 40%, 2-year OS rate was 16.7%. MS-PCR detection showed that the decitabine could significantly reduce the ID4 gene methylation level. It is concluded that decitabine can stabilize disease status of MDS patients, reduce blood transfusion dependence and improve the life quality of patients, and even some patients who transformed from MDS to leukemia achieved CR after treatment with decitabine. Decitabine can reduce the ID4 gene methylation level. The main adverse reaction of decitabine was myelosuppression, infection and so on. So the blood transfusions, antibiotics and other supportive treatments for these patients are needed. Most of patients well tolerate the adverse effects of decitabine after active symptomatic and supportive treatment. The efficacy and survival rate of patients in this study were similar to that of application of decitabine to treat MDS in other domestic studies.

[Application of multiplex nested RT-PCR to detecting 10 fusion genes related with MLL gene in myelodysplastic syndrome].

This study was aimed to investigate the clinical value of multiplex nested reverse transcription PCR (RT-PCR) in detecting MLL-related fusion genes in myelodysplastic syndrome (MDS). Ten MLL-related genes (dupMLL, MLL-ELL, MLL-ENL, MLL-AF6, MLL-AF9, MLL-AF10, MLL-AF17, MLL-CBP, MLL-AF1P, MLL-AF1Q) in 221 MDS cases were detected by multiplex nested RT-PCR. The results indicated that 20 patients were detected with positive result among 221 patients and the positive rate was 9.05%. The number of the positive cases and positive rates of the above mentioned 10 fusion genes were in order: 7 (3.16%), 2 (0.9%), 1 (0.45%), 1 (0.45%), 2 (0.9%), 2 (0.9%), 1 (0.45%), 2 (0.9%), 1 (0.45%), 1 (10.45%). It is concluded that the multiplex nested RT-PCR has been confirmed to be able to detect 10 fusion genes in MDS patients, which can provide important evidences for assessing diagnosis and treatment, and give related necessary information about minimal residual disease and its prognosis.

[Advances of treatment for extranodal NK/T-cell lymphoma --- review].

Extranodal NK/T-cell lymphoma is a rare pathological type, incidence of which is 2% - 10% of the primary NHL cases, showing a broad morphologic spectrum with frequent necrosis. Extranodal NK/T-cell lymphoma occurs more frequently in Asian population, especially in Southern China and Southeast Asian population. It is reported that the incidence of ENKL among lymphomas (1314 cases) in Asian countries was 4 times as much as the Western countries (22% vs. 5%) by the International Peripheral T-cell Lymphoma Project. Nasal NK/T-cell lymphoma most commonly occurred in the nasal cavity or other parts of the upper aerodigestive tract, and is highly associated with EBV (Epstein-Barr virus) infection, which is highly aggressive and its prognosis generally is poor. The mean survival time is about 12-38 months. It is important to accurately assess the patients prognosis for an optimal treatment. Localized disease (stage I and II) often has a relatively good prognosis through local radiation or combined therapy. There is a paucity of data to guide therapy in advanced disease. Generally, combining therapy is the most commonly selected approach for advanced disease. The conditions of the patients with advanced, relapsed or refractory diseases have been improved by high-intensity chemotherapy combined with radiation, and a regimen containing L-asparaginase L-Asp. Recently some studies have demonstrated promising outcomes in the selected cases by high-dose chemotherapy supplemented with auto-or allo-HSCT. Targeting therapy is also developing quickly. This current review mainly focuses on the advance of treatment for extranodal NK/T-cell lymphoma.

[Therapy and prognosis analysis of 47 patients with extranodal NK/T-cell lymphoma].

This study was purposed to explore the clinical characteristics, therapy and prognosis of patients with extranodal NK/T cell lymphoma (ENKL). 47 patients with ENKL from October 1995 to December 2008 in our hospital were analyzed retrospectively. The survival of patients was analyzed by using Kaplan-Meier methods, the prognosis of patients was evaluated by multivariate analysis using COX regression model. The clinical parameters used included CD56, Ann Arbor stage, international prognostic index (IPI) and B symptom. The results showed that the 2-year and 5-year overall survival (OS) rates were 91%, 71% respectively. Multivariant analysis by COX regression showed the CD56 and Ann Arbor stage were independent prognostic factors. Single factor analysis with staging in CHOP chemotherapy group indicated that more than stage IIIE (including IIIE) was a prognostic factor. Single factor analysis with B symptom showed that B symptom also was a prognostic factor. The cumulative survival rate of patients received radiotherapy alone was higher than that of patients got chemotherapy alone, its difference had a statistical significance, but there was no statistical significance between radiotherapy group and chemoradiotherapy group. It is concluded that CD56, Ann Arbor stage and B symptoms are prognostic factors. Radiotherapy alone is better than chemotherapy alone in therapy of ENKL. Chemotherapy combined with radiotherapy can not improve the survival. Improving the status of patients can make prognosis better.