Volume 99 ·  Issue 48 ·  2019

To analyze the copy number variation of CYP21A2 gene in 21-hydroxylase deficiency (21-OHD) patients, and identify the three copy repetition, single copy deletion of CYP21A2 gene and the type and proportion of CYP21A1P/CYP21A2 fused gene in 21-OHD patients.

A total of 424 patients (140 males and 284 females) with 21-OHD who visited Peking Union Medical College Hospital from January 2015 to January 2018 were enrolled and the average age was (17.1±12.4) years. All clinical and biochemical data were collected. DNAs were extracted from peripheral blood leukocytes, and CYP21A2 gene mutation and copy number variation were detected by Sanger sequencing and multiple ligation probe amplification (MLPA).

Of 424 21-OHD patients, 287 (67.7%) had two copies of CYP21A2 gene, 137 (32.3%) had copy number variation, of which 1 patients (0.2%) had 3 copies of CYP21A2 gene and 136 (32.1%) were carriers of large deletion/rearrangement mutation of CYP21A2 gene. Three pathogenic mutations including a truncated Q319X protein mutation were detected in the patient with 3 copies of CYP21A2 gene. Of 136 patients with large deletion/rearrangement mutation of CYP21A2 gene, 82 (60.3%) carried fused CYP21A1P/CYP21A2 gene, and the remaining 54 harbored the one allele deletion of CYP21A2. The most common types of fused CYP21A1P/CYP21A2 gene were CH-5, CH-1 and CH-2, with the frequency being 31.7% (26 cases), 26.8% (22 cases) and 19.5% (16 cases), respectively, and followed by CH-4 and CH-7, with the incidence being 8.5% (7 cases) and 4.9% (4 cases), respectively. In addition, two cases of CH-3, CH-6 and CH-8 and one case of CH-9 were detected.

This is the first study to detect the occurrence of CYP21A2 gene copy number variation and fused CYP21A1P/CYP21A2 gene in a large cohort of 21-OHD patients. The number of CYP21A2 gene copies in 21-OHD patients includes 2 copies, 1 copy deletion and 3 copies duplication. One copy deletion of CYP21A2 includes one allele deletion of CYP21A2 gene and fused CYP21A1P/CYP21A2 gene. In patients with 3 copies of CYP21A2 gene, pathogenic mutations should be verified in all 3 copies of CYP21A2 gene to make the precise diagnosis. Therefore, the accurate molecular diagnosis of 21-OHD patients should take both genotype and copy number variation of CYP21A2 into account.

To investigate the safety and myocardial protective effect of del Nido cardioplegia in coronary artery bypass grafting combined with heart valve replacement in adults.

From April 2018 to April 2019, 84 patients undergoing coronary artery bypass grafting combined with heart valve replacement under cardiopulmonary bypass (CPB) in the Second Affiliated Hospital of Nantong University and Shanghai Chest Hospital were selected, and divided into del Nido cardioplegia (n=44) group and Thomas cardioplegia (n=40) group using a random number table. The concentration of troponin was measured before anesthesia induction and 2, 12 and 24 hours after the termination of CPB, respectively.

The preoperative characteristics of each group were similar, including age, underlying diseases, and body surface area (all P>0.05). The frequency of intraoperative cardioplegia infusion in the del Nido group was significantly lower than that in the Thomas group (P=0.03). Compared with the del Nido group, troponin concentration in the Thomas group increased significantly at 12 hours [(4.59±0.76) μg/L vs (2.20±0.53) μg/L, t=-16.653, P<0.001] and 24 hours [(6.70±0.97) μg/L vs (1.96±0.58) μg/L, t=-26.975, P<0.001] after CPB. No statistically significant differences were found in the postoperative recovery of cardiac autonomic rhythm (P=0.887), cardiac defibrillation (P=0.880), positive inotropic drugs (P=0.163) and left ventricular ejection fraction (P=0.338) between the two groups.

It is safe to use del Nido cardioplegia in coronary artery bypass grafting combined with heart valve replacement in adults, which can reduce the frequency of cardioplegia infusion and simplify the surgical procedure. The trend of troponin concentration suggests that del Nido cardioplegia has a better myocardial protective effect.

To investigate the value of rapid on-site evaluation (ROSE) of bronchoscopy in the diagnosis of pulmonary complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

A retrospective analysis was conducted on the diagnosis and treatment process before and after ROSE examination of 12 patients with pulmonary complications after allo-HSCT who were admitted to the Department of hematology, Tianjin First Central Hospital from January 2017 to June 2019. The diagnostic accuracy of the ROSE was evaluated by comparing the initial diagnosis of ROSE with the final clinical diagnosis. At the same time, the safety of ROSE examination was evaluated and two typical cases were shared.

In the 12 transbronchial lung biopsies, there were 5 cases of organizing pneumonia, 3 cases of bronchiolitis obliterans with organizing pneumonia, 1 case of pulmonary fibrosis, 1 case of acute fibrinous and organizing pneumonia, 1 case of pseudomembranous tracheobronchial aspergillosis and 1 case of uncertain diagnosis evaluated by ROSE. Compared with the final clinical diagnosis, there were 10 cases of accurate diagnosis made by ROSE (10/12). All patients were well tolerant to the operation of bronchoscopy. There was only a small amount of bleeding observed during the operation, without pneumothorax and hemoptysis. And no complications related to ROSE occurred. According to the initial diagnosis of ROSE, 10 cases of non-infectious pulmonary complications were treated with methylprednisolone or other immunosuppressive agents and 1 case of Aspergillus infection was given antifungal therapy. Seven patients with non-infectious pulmonary complications improved after treatment. One patient obtained uncertain diagnosis by ROSE was later diagnosed with virus infection by next generation sequencing technology and improved after treatment with foscarnet sodium and immunoglobulin. As of June 30, 2019, 7 patients improved and 5 died.

ROSE has the advantages of diagnostic accuracy and rapidity, and is very suitable for patients with critical illness after hematopoietic stem cell transplantation, who are in urgent need of definite diagnosis and prompt treatment, which is beneficial to improve the prognosis of patients.

To investigate the efficacy, safety and compliance of dual therapy for the first-line eradication of Helicobacter pylori infection through a prospective, single-center and open-label cohort study.

From March 2014 to September 2018, 200 naïve patients with Helicobacter pylori infection and dyspepsia received 14-day dual therapy (rabeprazole 10mg and amoxicillin 500 mg, four times daily orally). Safety and compliance were assessed 1-3 days after eradication. The therapeutic outcome was determined by ¹³C-urea breath test 4-8 weeks after eradication. Some patients underwent strain culture, antibiotic sensitivity testing and CYP2C19 polymorphism assay.

The eradication rates of dual therapy: intention-to-treat analysis 87.5% (95% confidence interval 82.5%-91.5%), modified intention-to-treat analysis 90.2% (86.1%-94.3%) and per-protocol analysis 91.0% (86.3%-94.7). 21.2% of patients had adverse reactions, the majority were mild to moderate, and only 1.5% of patients discontinued medication because of intolerance to adverse reactions. Patients with good compliance accounted for 96.0%. Variate analyses showed that poor compliance and amoxicillin resistance were the independent risk factors for eradication failure.

14-day dual therapy with rabeprazole and amoxicillin (four times daily) achieved good efficacy, safety and compliance for the first-line eradication of Helicobacter pylori infection.

To evaluate the clinical outcomes in patients with relapsed or refractory peripheral T-cell lymphoma (PTCL) who had undergone allogeneic hematological stem cell transplantation (allo-HSCT).

From June 2007 to June 2017, the clinical data of PTCL patients who underwent HSCT from eight hospitals were assessed retrospectively.

There were 23 patients diagnosed as relapsed or refractory PTCL with chemoresistance who underwent allo-HSCT. Among these patients, 18 were identified as progressive disease (PD) status and 5 patients as stable disease (SD) status before allo-HSCT. Seventeen patients received allo-HSCT from matched sibling donor (MSD),2 patients from matched unrelated donor and 4 patients from related haplo-identical donor (HD). After a median follow-up of 29 months, 21 patients survived longer than 28 days after allo-HSCT. Hematopoietic reconstitution was achieved in 20 of the 21 patients. The median time of myeloid and platelet engraftment were+13 (9-22) d and+16(10-38) d, respectively. The 100-d treatment-related mortality rate was 13.1%. Acute GVHD occurred in 11(47.8%) patients at a median time of 22(6-82) d after transplantation. Grade Ⅱ~Ⅳ aGVHD occurred in 6 patients. Chronic GVHD occurred in 10 patients at a median of 7.9 (3.5-27) months. After a median follow-up of 29 months, 13 patients died after HSCT. Four of them died of complications associated with allo-HSCT, and other 9 patients died of the primary lymphoma. The 3-years cumulative overall survival (OS) and progress-free survival (PFS) were 43.03% (95%CI: 29.79-69.16) and 39.13% (95%CI: 23.50-65.14), respectively. No significant difference was found in the 3-year PFS between patients with PD status and SD status before allo-HSCT (P=0.133).

Allo-HSCT can be a promising treatment for relapsed or refractory PTCL with chemoresistance.

To analyze the clinical characteristics and prognostic factors of patients with Hodgkin′s lymphoma(HL).

The clinical data of HL patients who were newly treated in Guizhou Cancer Hospital from August 2007 to March 2019 was retrospectively collected, and the efficacy and prognostic factors were analyzed.

The clinical data of a total of 222 patients were analyzed in this study. The 5-year progression-free survival (PFS) and overall survival (OS)rate of them were 82.8% and 81.3%, respectively. The 5-year PFS rate and OS rate of early (stage Ⅰ-Ⅱ) HL were 87.3% and 86.1%, respectively, and the 5-year PFS rate and OS rate of progressive (stage Ⅲ-Ⅳ) HL were 77.9% and 76.3%, respectively. Among the 118 patients with early Hodgkin′s lymphoma, the complete remission(CR) rate of chemotherapy alone was 55.6%(15/27), and chemotherapy plus radiotherapy was 86.8% (79/91), the difference between which was statistically significant (P<0.05). Compared with chemotherapy plus radiotherapy in early stage patients, 5-year PFS (93.0%) and 5-year OS (92.0%) rate in patients with chemotherapy plus radiotherapy were better than those with chemotherapy alone which were 63.7% and 62.1%, respectively. Multivariate analysis showed that age, LDH, ABVD cycle number and chemoradiotherapy were independent prognostic factors for 5-year OS and PFS rate in HL patients. The adverse reactions were increased level Ⅰ-Ⅱ aminotransferase with an incidence of 47.7% (106/222), decreased level Ⅰ-Ⅱ neutrophils with an absolute value of 54.1% (120/222), and decreased level Ⅲ-Ⅳ neutrophils with an absolute value of 45.9% (102/222). No adverse cardiac and pulmonary reactions or secondary tumors associated with chemotherapy was found in all patients.

HL is a type of malignant tumor with good prognosis, and the short-term and long-term efficacy of chemotherapy combined with radiotherapy in early patients is better than that of chemotherapy alone. Age, LDH, ABVD cycle number and chemoradiotherapy are associated with prognosis in patients with Hodgkin′s lymphoma. Adverse reactions can be tolerated.

To explore the clinical efficacy of damage control orthopaedics in treatment of patients with severe traumatic brain injury combined with limb fracture.

Total 149 patients with sTBI combined with limb fracture treated in Emergency Surgery Department of Shandong University Qilu Hospital from January, 2011 to December, 2018 were collected. Patients that were treated with immediate definitive fixation before March 31st, 2013 were included into the routine treatment group (group A, 47 cases), and that were treated with damage control orthopaedics (DCO) after April 1st, 2013 were included into the DCO group (group B, 102 cases). The clinical data during hospitalization and prognosis 3 months after injury was analyzed retrospectively. Then according to the risk of postoperative ICP increase, the two groups were divided into low risk subgroup and high risk subgroup. The postoperative ICP, length of stay and GOS score were statistically analyzed.

There were no statistically significant differences in sex, age, preoperative GCS score, imaging type of lesion and initial intraoperative ICP between the two groups. The postoperative ICP and incidence of coagulation dysfunction were significantly higher in group A [(17.1±4.6) mmHg, 29.8%] than that in group B[(15.0±4.3) mmHg, 13.7%] separately(P<0.05), and there was no significant difference of length of stay and GOS score between the two groups (P=0.475 and 0.097, respectively). As for the subgroup with low risk of postoperative ICP increase, there was no significant difference in postoperative ICP and GOS score between group A and B, and the length of stay of group B was significantly longer than that of group A (P<0.05). As for the subgroup with high risk of postoperative ICP increase, there was no significant difference in the length of stay between group A and B (P=0.667), and for group A the postoperative ICP was higher and GOS score was lower than that of group B (P<0.05).

For patients with sTBI combined with limb fracture, the application of DCO should be based on the severity of traumatic brain injury. For patients with high risk of postoperative ICP increase, DCO can significantly improve the prognosis of patients.

To analyze the causes of complications after endoscopic transsphenoidal pituitary adenoma resection and summarize the corresponding prevention and treatment measures.

The pathological data of 120 patients with pituitary adenoma in hospital were analyzed retrospectively. All patients were treated surgically, five kinds of common complications were collected, the risk factors of complications were statistically analyzed, the causes of postoperative complications were determined, and then the specific prevention and treatment strategies were summarized.

Long operation time (P<0.05), hypopituitarism before operation (P<0.01) and large tumor (P<0.01) were the main inducing factors of CSF rhinorrhea after operation; long operation time (OR=1.394, P<0.01), lumbar cistern puncture (OR=1.531, P<0.01) and CSF rhinorrhea after operation (OR=3.861, P<0.01) were the independent risk factors of infection; Age, large tumor and preoperative hypophysis are the main factors of postoperative hypophysis; Postoperative hypophysis and intraoperative cerebrospinal fluid rhinorrhea are the main factors of postoperative diabetes insipidus; Postoperative hypophysis and giant adenoma are the main factors of postoperative hyponatremia.

Endoscopic transnasal transsphenoidal resection of pituitary adenoma is the most minimally invasive method at present. There are still risk factors for postoperative complications. Targeted prevention of risk factors for complications and corresponding interventions can effectively reduce the incidence of postoperative complications of pituitary adenoma

To investigate the effect of silencing fatty acid binding protein 3 (FABP3) gene on lipopolysaccharide (LPS)-induced apoptosis and endoplasmic reticulum stress in alveolar epithelial cells A549.

According to the processing method, A549 cells were divided into control group(A549 cells cultured for 24 h), LPS group (10 mg/L LPS treated A549 cells for 24 h), LPS+si-con group (10 mg/L LPS was used to treat A549 cells transfected with si-con for 24 h) and LPS+si-FABP3 group (10 mg/L LPS was used to treat A549 cells transfected with si-FABP3 for 24 h). Then quantitative real-time PCR was used to detect the level of FABP3, methylthiazoletrazolium was used to detect the cell proliferation, flow cytometry was used to detect the apoptosis, and Western Blot was used to detect the levels of FABP3, CyclinD1, cleaved-caspase-3, GRP78, ATF4, CHOP, cleaved-caspase-12 and p-Akt and PI3Kp110α protein expression. Enzyme-linked immunosorbent assay was used to detect the levels of IL-6, IL-8 and TNF-αlevels.

In the LPS group, FABP3 protein level (1.00±0.09) and mRNA (2.15±0.22), apoptosis rate [(26.1±2.6)%], inflammatory factor IL-6 [(554.4±55.4) ng/L], IL-8 [(389.3±38.5) ng/L] and TNF-α [(601.3±60.0) ng/L], cleaved-caspase-3 (1.00±0.11), GRP78 (1.05±0.11), ATF4 (1.20±0.12)), CHOP (1.05±0.10), cleaved-caspase-12 (1.10±0.11), p-Akt (0.88±0.08) and PI3Kp110α (0.75±0.08) protein levels were significantly higher than the control group [(0.53±0.05), (1.00±0.10), (4.5±0.5)%, (75.4±7.5) ng/L, (25.2±2.5) ng/L, (66.5±6.7) ng/L, (0.34±0.05), (0.35±0.05), (0.43±0.05), (0.37±0.04), (0.45±0.05), (0.16±0.04), (0.35±0.05)] (allP<0.05). Cell viability [(50.1±5.4)%] and CyclinD1 protein level (0.40±0.05) in LPS group were significantly lower than those in the control group [(100.1±12.4)%, (1.25±0.12)] (bothP<0.05). Cell viability [(89.1±8.5)%] and CyclinD1 protein level (1.15±0.11) in LPS+si-FABP3 group were significantly higher than those in LPS+si-con group [(53.1±5.4)%, (0.42±0.05)] (bothP<0.05). Apoptosis rate [(10.5±1.1)%], IL-6[(301.3±30.0) ng/L], IL-8[(189.4±19.0) ng/L], TNF-α [(400.1±40.1) ng/L], cleaved-caspase-3 (0.45±0.05), GRP78 (0.48±0.05), ATF4 (0.60±0.06), CHOP (0.55±0.05), cleaved-caspase-12 (0.60±0.06), p-Akt (0.50±0.05) and PI3Kp110α(0.45±0.05) in LPS+si-FABP3 group were significantly lower than those in LPS+si-con group [(28.1±2.8)%, (536.3±53.6) ng/L, (400.2±40.2) ng/L, (623.1±62.3) ng/L, (0.96±0.10), (1.02±0.10), (1.15±0.12), (1.10±0.11), (1.15±0.12), (0.90±0.09), (0.72±0.07)] (allP<0.05).

Silencing FABP3 gene can inhibit LPS-induced alveolar epithelial cell apoptosis and endoplasmic reticulum stress, which may act by inhibiting the PI3K/Akt signaling pathway.

To investigate the effects of miR-106a-5p on the proliferation and migration of vascular endothelial cells, and the possible target gene of miR-106a-5p in endothelial cells.

Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and the expression of mir-106a-5p in HUVECs was detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). Cell counting kit-8 (CCK-8) and wound healing assays were used to detected the proliferation and migration of HUVECs respectively.Dual luciferase repoter assay was used to identify the possible target gene of miR-106a-5p——STAT3.

mir-106a-5p inhibited the proliferation function of HUVECs (F=13.62, P<0.01). And mir-106a-5p expressed the migration of HUVEC,the closure rate in the mimic group was reduced after 12 h and 24 h of scratching (49.93/31.31) (χ²=8.240, P<0.05), (78.87/44.80) (χ²=10.50, P<0.01). The direct target gene of mir-106a-5p was STAT3, and miR-106a-5p regulated the expression of STAT3 through post-transcriptionally controlling.

mir-106a-5p could inhibit proliferation and migration of HUVEC, and the possible target gene was STAT3.

To evaluate the prognostic impact of neuromuscular blocking agents (NMBA) on patients with acute respiratory distress syndrome (ARDS).

Online search of MEDLINE, Embase, Web of Science, CNKI, CBM and other Chinese databases for randomized controlled trials (RCTs) of NMBA in patients with ARDS from January 1994 to June 2019 was done, and literature was selected according to inclusion and exclusion criteria. The patients were divided into NMBA group and non-NMBA group according to whether NMBA was adopted or not. The prognostic indicators (ICU mortality, 28 d mortality, 90 d mortality) and NMBA-related complications (ICU acquired muscle weakness, barometric injury, pneumothorax) of the patients in the two groups were mainly analyzed. Meta-analysis of the data was performed using RevMan 5.0 software.

A total of 6 RCTs were included, and 1 502 patients were enrolled, including 761 in the NMBA group and 741 in the no-NMBA group. The 90-day mortality in the NMBA group and no-NMBA group were 38.8% and 42.6%, OR=0.87 (95%CI: 0.70-1.07, P=0.190); the 28-day mortality rates were 32.5% and 36.5%, OR=0.71 (95%CI: 0.45-1.11, P=0.130); ICU mortality rates were 31.8% and 43.8%, OR=0.60 (95%CI: 0.41-0.88, P=0.009).

NMBA can reduce the ICU mortality of moderate to severe ARDS patients, but not reduce 28-day and 90-day mortality.