Determination of suspended erythrocyte volume range and establishment of its internal control standard / 中国输血杂志

【Objective】 To determine the volume range of suspended erythrocyte and establish its internal control standard. 【Methods】 The theoretical value of suspended erythrocyte volume was calculated according to the screening criteria of healthy blood donors and Quality Requirements for Whole Blood and Blood Components. A total of 2 410 bags of 1 U and 2 U suspended erythrocyte were randomly selected and weighed, and the volume range were formulated by ±2S and ±10% respectively and then compared to determine the volume range in line with the actual situation of our center. 【Results】 The theoretical volume range of 1 U and 2 U suspended erythrocyte were 117-160 mL vs 234-320 mL, and the actual volume range were 142-180 mL vs 276-393 mL. The volume range of 1 U and 2 U suspended erythrocyte formulated by ±2S were 145-181 mL vs 298-358 mL, and by ±10% were 147-179 mL vs 295-361 mL. The hematocrit and hemoglobin content of suspended erythrocyte within the actual volume range met the quality requirements. There were fluctuations in the volume of suspended erythrocyte from different regions. 【Conclusion】 Based on the actual situation of our center and the sampling results of suspended erythrocytes in recent two years, 163 mL±10% and 328 mL±10% were determined as the internal control standards of 1 U and 2 U suspended erythrocyte, respectively. Blood centers should establish accurate and feasible standard of suspended erythrocyte according to the actual situation.

Continuous and Intermittent Exercise Upregulates PGC-1α/ Irisin / BDNF Expression to Improve Obesity-induced Cognitive Impairment in Mice / 中国生物化学与分子生物学报

This study aimed to investigate the effects of exercise on prefrontal PGC-1α, Irisin, BDNF, oxidative stress, inflammation, and cognitive function in high-fat diet-induced obese mice, which may provide experimental evidence of exercise rehabilitation methods and target screening for obesity. Three-month-old male C57BL/ 6J wild-type mice were randomly divided into four groups: control, high-fat diet, high-fat diet with moderate intensity continuous training, and high-fat diet with high-intensity interval training group, with 10 mice in each group. The mice in the high-fat diet with moderate intensity continuous training group and the high-fat diet with high-intensity interval training group received 8 weeks of moderate-intensity continuous training or high-intensity interval training after 12-week high-fat feeding. Behavioral results showed that compared with the control group, reaction time of adhesive removal test was significantly increased (P<0. 01), and spontaneous alternation rate in Y-maze test and exploration time in the novel object recognition test were significantly decreased (P < 0. 01) in the high-fat diet group, indicating that high-fat diet led to cognitive dysfunction in mice. Results showed that compared with the control group, Nissl bodies dissolution and apoptosis were significantly increased (P<0. 01), levels of PGC-1α, Irisin, BDNF, IL-10, and T-SOD were significantly deceased (P<0. 05, P<0. 01), levels of IL-1β, TNF-α, NF-κB, cleaved Caspase-3, Bax/ Bcl-2, ROS, and MDA were significantly increased in the prefrontal lobe (P<0. 01), indicating that high-fat diet induced excessive inflammation, oxidative stress, and apoptosis, which were conducive to prefrontal lobe damage. Compared with the high-fat diet group, moderate intensity continuous or high-intensity interval training decresed reaction time of adhesive removal test (P<0. 01), increased spontaneous alternation rate of Y-maze test and exploration time of novel object recognition test (P<0. 05, P<0. 01), indicating that both continuous and interval training improved cognitive function in obese mice; meanwhile, Nissl bodies and levels of PGC-1α, Irisin, BDNF, IL-10, and T-SOD were significantly increased (P<0. 05, P<0. 01), and apoptosis and levels of IL-1β, TNF-α, NF-κB, cleaved Caspase-3, Bax/ Bcl-2, ROS, and MDA were significantly reduced in the prefrontal lobe (P<0. 01), indicating that both continuous and interval training alleviated obesity-induced inflammation, oxidative stress, and apoptosis in the prefrontal lobe. Both continuous and interval training significantly upregulated PGC-1α/ Irisin/ BDNF expression in the prefrontal lobe of obese mice, inhibited oxidative stress and inflammation, reduced apoptosis, and resulted in alleviating obesity-induced prefrontal lobe damage and cognitive dysfunction; morevover, interval training better than continuous.

The effect of platelet infusion on the repair of bone marrow hematopoietic niche damage in mice induced by (60)Co radiation and hematopoietic reconstruction after bone marrow transplantation / 中华血液学杂志

Objective: To observe the effect of platelets on hematopoietic stem cell (HSCs) implantation in mice with radiation-induced bone marrow injury and bone marrow transplantation models. Methods: ①Male C57BL/6 mice were divided into a single irradiation group and a radiation infusion group after receiving (60)Co semimyeloablative irradiation for 18-10 weeks. The irradiation infusion group received 1×10(8) platelets expressing GFP fluorescent protein. ② The allogeneic bone marrow transplantation model was established. The experimental groups included the simple transplantation group (BMT) and the transplantation infusion group (BMT+PLT). The BMT group was infused through the tail vein only 5 × 10(6) bone marrow cells, the BMT+PLT group needs to be infused with bone marrow cells at the same time 1× 10(8) platelets. ③ Test indicators included peripheral blood cell and bone marrow cell counts, flow cytometry to detect the proportion of hematopoietic stem cell (HSC) and hematopoietic progenitor cells, bone marrow cell proliferation and apoptosis, and pathological observation of vascular niche damage and repair. Results: ①On the 3rd, 7th, 14(th), and 21st days after irradiation, the bone marrow cell count of the infusion group was higher than that in the single irradiation group (P<0.05), and the peripheral blood cell count was also higher. A statistically significant difference was found between the white blood cell count on the 21st day and the platelet count on the 7th day (P<0.05). In the observation cycle, the percentage of bone marrow cell proliferation in the infusion group was higher, while the percentage of apoptosis was lower. ② The results of bone tissue immunofluorescence after irradiation showed that the continuity of hematopoietic niche with red fluorescence was better in the irradiation infusion group. ③The chimerism percentage in the BMT+PLT group was always higher than that in the BMT group after transplantation.④ The BMT+PLT group had higher bone marrow cell count and percentage of bone marrow cell proliferation on the 7th and 28th day after transplantation than that in the BMT group, and the percentage of bone marrow cell apoptosis on the 14th day was lower than that in the BMT group (P<0.05). After the 14th day, the percentage of stem progenitor cells in the bone marrow cells of mice was higher than that in the BMT group (P<0.05). ⑤The immunohistochemical results of bone marrow tissue showed that the continuity of vascular endothelium in the BMT+PLT group was better than that in the BMT group. Conclusion: Platelet transfusion can alleviate the injury of vascular niche, promotes HSC homing, and is beneficial to hematopoietic reconstruction.

Diagnosis and management of orbital and cranial complications of pediatric acute rhinosinusitis / 中华耳鼻咽喉头颈外科杂志

Objective: To review the clinical characteristics, to illustrate diagnosis and management experience of orbital and cranial complications of pediatric acute rhinosinusitis. Methods: The clinical data of 24 children with orbital and cranial complications of acute rhinosinusitis who received endoscopic sinus surgery combined with drug treatment in Beijing Children's Hospital from January 2017 to December 2021 were retrospectively reviewed. There were 19 boys and 5 girls. The age varied from 13 to 159 months, with a median 47.5 months. The following diagnoses were obtained: 12 isolated subperiosteal orbital abscess, 2 associated with preseptal abscess, 2 associated with intraorbital abscess, 7 associated with optic neuritis, and 1 associated with septic cavernous sinus thrombosis. Clinical characteristics, organism isolated and outcomes were analyzed through descriptive methods. Results: All 24 patients presented with fever; 9 presented with nasal congestion and purulent discharge. The clinical manifestations of orbital infection included orbital edema, pain, proptosis and displacement of globe in all patients, while visual impairment was recognized in 7 children. Purulent drainage was cultured in 17 patients, among which 12 were positive. All patients underwent nasal endoscopic surgical interventions uneventfully, excluding one patient who required a second surgical procedure. Follow-up period ranged from 5 to 64 months. All patients resolved fully, with the exception of 2 children who got permanent blindness with visual loss preoperative. There was no recurrence or death. Conclusions: Orbital and cranial complications of pediatric acute rhinosinusitis could be severe with an occult onset. For patients with vison impairment, any signs of intracranial complications and a lack of response to conservative management, an urgent endoscopic intervention is needed.

Evaluation of peripheral blood T-lymphocyte subpopulations features in patients with hepatitis B virus-related acute-on-chronic liver failure based on single-cell sequencing technology / 中华肝脏病杂志

Objective: T lymphocyte exhaustion is an important component of immune dysfunction. Therefore, exploring peripheral blood-exhausted T lymphocyte features in patients with hepatitis B virus-related acute-on-chronic liver failure may provide potential therapeutic target molecules for ACLF immune dysfunction. Methods: Six cases with HBV-ACLF and three healthy controls were selected for T-cell heterogeneity detection using the single-cell RNA sequencing method. In addition, exhausted T lymphocyte subpopulations were screened to analyze their gene expression features, and their developmental trajectories quasi-timing. An independent sample t-test was used to compare the samples between the two groups. Results: Peripheral blood T lymphocytes in HBV-ACLF patients had different differentiation trajectories with different features distinct into eight subpopulations. Among them, the CD4(+)TIGIT(+) subsets (P = 0.007) and CD8(+)LAG3(+) (P = 0.010) subsets with highly exhausted genes were significantly higher than those in healthy controls. Quasi-time analysis showed that CD4(+)TIGIT(+) and CD8(+)LAG3(+) subsets appeared in the late stage of T lymphocyte differentiation, suggesting the transition of T lymphocyte from naïve-effector-exhausted during ACLF pathogenesis. Conclusion: There is heterogeneity in peripheral blood T lymphocyte differentiation in patients with HBV-ACLF, and the number of exhausted T cells featured by CD4(+)TIGIT(+)T cell and CD8(+)LAG3(+) T cell subsets increases significantly, suggesting that T lymphocyte immune exhaustion is involved in the immune dysfunction of HBV-ACLF, thereby identifying potential effective target molecules for improving ACLF patients' immune function.

Clinicopathologic characteristics and prognostic analysis of testicular diffuse large B-cell lymphoma / 中华血液学杂志

Objective: To analyze the clinicopathologic characteristics and prognosis of testicular diffuse large B-cell lymphoma (DLBCL) . Methods: A retrospective analysis was performed on 68 patients with testicular DLBCL admitted to Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine from October 2001 to April 2020. The gene mutation profile was evaluated by targeted sequencing (55 lymphoma-related genes) , and prognostic factors were analyzed. Results: A total of 68 patients were included, of whom 45 (66.2% ) had primary testicular DLBCL and 23 (33.8% ) had secondary testicular DLBCL. The proportion of secondary testicular DLBCL patients with Ann Arbor stage Ⅲ-Ⅳ (P<0.001) , elevated LDH (P<0.001) , ECOG score ≥ 2 points (P=0.005) , and IPI score 3-5 points (P<0.001) is higher than that of primary testicular DLBCL patients. Sixty-two (91% ) patients received rituximab in combination with cyclophosphamide, adriamycin, vincristine, and prednisone (R-CHOP) -based first-line regimen, whereas 54 cases (79% ) underwent orchiectomy prior to chemotherapy. Patients with secondary testicular DLBCL had a lower estimated 5-year progression-free survival (PFS) rate (16.5% vs 68.1% , P<0.001) and 5-year overall survival (OS) rate (63.4% vs 74.9% , P=0.008) than those with primary testicular DLBCL, and their complete remission rate (57% vs 91% , P=0.003) was also lower than that of primary testicular DLBCL. The ECOG scores of ≥2 (PFS: P=0.018; OS: P<0.001) , Ann Arbor stages Ⅲ-Ⅳ (PFS: P<0.001; OS: P=0.018) , increased LDH levels (PFS: P=0.015; OS: P=0.006) , and multiple extra-nodal involvements (PFS: P<0.001; OS: P=0.013) were poor prognostic factors in testicular DLBCL. Targeted sequencing data in 20 patients with testicular DLBCL showed that the mutation frequencies of ≥20% were PIM1 (12 cases, 60% ) , MYD88 (11 cases, 55% ) , CD79B (9 cases, 45% ) , CREBBP (5 cases, 25% ) , KMT2D (5 cases, 25% ) , ATM (4 cases, 20% ) , and BTG2 (4 cases, 20% ) . The frequency of mutations in KMT2D in patients with secondary testicular DLBCL was higher than that in patients with primary testicular DLBCL (66.7% vs 7.1% , P=0.014) and was associated with a lower 5-year PFS rate in patients with testicular DLBCL (P=0.019) . Conclusion: Patients with secondary testicular DLBCL had worse PFS and OS than those with primary testicular DLBCL. The ECOG scores of ≥2, Ann Arbor stages Ⅲ-Ⅳ, increased LDH levels, and multiple extra-nodal involvements were poor prognostic factors in testicular DLBCL. PIM1, MYD88, CD79B, CREBBP, KMT2D, ATM, and BTG2 were commonly mutated genes in testicular DLBCL, and the prognosis of patients with KMT2D mutations was poor.

Effect of diosgenin on mTOR/FASN/HIF-1α/VEGFA expression in rats with non-alcoholic fatty liver disease / 中国中药杂志

The present study aimed to investigate the effect of diosgenin on mammalian target of rapamycin(mTOR), fatty acid synthase(FASN), hypoxia inducible factor-1α(HIF-1α), and vascular endothelial growth factor A(VEGFA) expression in liver tissues of rats with non-alcoholic fatty liver disease(NAFLD) and explore the mechanism of diosgenin on lipogenesis and inflammation in NAFLD. Forty male SD rats were divided into a normal group(n=8) fed on the normal diet and an experimental group(n=32) fed on the high-fat diet(HFD) for the induction of the NAFLD model. After modeling, the rats in the experimental group were randomly divided into an HFD group, a low-dose diosgenin group(150 mg·kg~(-1)·d~(-1)), a high-dose diosgenin group(300 mg·kg~(-1)·d~(-1)), and a simvastatin group(4 mg·kg~(-1)·d~(-1)), with eight rats in each group. The drugs were continuously given by gavage for eight weeks. The levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), alanine transaminase(ALT), and aspartate transaminase(AST) in the serum were detected by the biochemical method. The content of TG and TC in the liver was detected by the enzyme method. Enzyme-linked immunosorbent assay(ELISA) was used to measure interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α) in the serum. Lipid accumulation in the liver was detected by oil red O staining. Pathological changes of liver tissues were detected by hematoxylin-eosin(HE) staining. The mRNA and protein expression levels of mTOR, FASN, HIF-1α, and VEGFA in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction(PCR) and Western blot, respectively. Compared with the normal group, the HFD group showed elevated body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1β, and TNF-α(P<0.01), increased lipid accumulation in the liver(P<0.01), obvious liver steatosis, up-regulated mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.01), and increased protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). Compared with the HFD group, the groups with drug treatment showed lowered body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1β, and TNF-α(P<0.05, P<0.01), reduced lipid accumulation in the liver(P<0.01), improved liver steatosis, decreased mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.05, P<0.01), and declining protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). The therapeutic effect of the high-dose diosgenin group was superior to that of the low-dose diosgenin group and the simvastatin group. Diosgenin may reduce liver lipid synthesis and inflammation and potentiate by down-regulating the mTOR, FASN, HIF-1α, and VEGFA expression, playing an active role in preventing and treating NAFLD.

Mechanism of heat-clearing prescriptions in alleviating type 2 diabetes mellitus:a review / 中国中药杂志

Type 2 diabetes mellitus(T2DM), a common chronic metabolic disease, is often accompanied by internal heat syndrome. Heat-clearing prescriptions are widely used to treat different heat syndromes of T2DM from the aspects of clearing stagnant heat, excess heat, damp heat, phlegm heat, and heat toxin, demonstrating remarkable effects. The mechanism of blood sugar-lowering agents has always been a hotspot of research. Recently, the basic studies of heat-clearing prescriptions from different perspectives have been increasing year by year. To clarify the mechanisms of heat-clearing prescriptions and find specific mechanisms, we systematically reviewed the basic studies of heat-clearing prescriptions commonly used for the treatment of T2DM in the past decade, intending to provide a reference for related research.

Diosgenin alleviates NAFLD induced by a high-fat diet in rats via mTOR/SREBP-1c/HSP60/MCAD/SCAD signaling pathway / 中国中药杂志

This study aims to observe the effects of diosgenin on the expression of mammalian target of rapamycin(mTOR), sterol regulatory element-binding protein-1c(SREBP-1c), heat shock protein 60(HSP60), medium-chain acyl-CoA dehydrogenase(MCAD), and short-chain acyl-CoA dehydrogenase(SCAD) in the liver tissue of the rat model of non-alcoholic fatty liver disease(NAFLD) and explore the mechanism of diosgenin in alleviating NAFLD. Forty male SD rats were randomized into five groups: a control group, a model group, low-(150 mg·kg~(-1)·d~(-1)) and high-dose(300 mg·kg~(-1)·d~(-1)) diosgenin groups, and a simvastatin(4 mg·kg~(-1)·d~(-1)) group. The rats in the control group were fed with a normal diet, while those in the other four groups were fed with a high-fat diet. After feeding for 8 weeks, the body weight of rats in the high-fat diet groups increased significantly. After that, the rats were administrated with the corresponding dose of diosgenin or simvastatin by gavage every day for 8 weeks. The levels of triglyceride(TG), total cholesterol(TC), alanine transaminase(ALT), and aspartate transaminase(AST) in the serum were determined by the biochemical method. The levels of TG and TC in the liver were measured by the enzyme method. Oil-red O staining was employed to detect the lipid accumulation, and hematoxylin-eosin(HE) staining to detect the pathological changes in the liver tissue. The mRNA and protein levels of mTOR, SREBP-1c, HSP60, MCAD, and SCAD in the liver tissue of rats were determined by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. Compared with the control group, the model group showed increased body weight, food uptake, liver index, TG, TC, ALT, and AST levels in the serum, TG and TC levels in the liver, lipid deposition in the liver, obvious hepatic steatosis, up-regulated mRNA and protein expression levels of mTOR and SREBP-1c, and down-regulated mRNA and protein expression levels of HSP60, MCAD, and SCAD. Compared with the model group, the rats in each treatment group showed obviously decreased body weight, food uptake, liver index, TG, TC, ALT, and AST levels in the serum, TG and TC levels in the liver, lessened lipid deposition in the liver, ameliorated hepatic steatosis, down-regulated mRNA and protein le-vels of mTOR and SREBP-1c, and up-regulated mRNA and protein levels of HSP60, MCAD, and SCAD. The high-dose diosgenin outperformed the low-dose diosgenin and simvastatin. Diosgenin may prevent and treat NAFLD by inhibiting the expression of mTOR and SREBP-1c and promoting the expression of HSP60, MCAD, and SCAD to reduce lipid synthesis, improving mitochondrial function, and promoting fatty acid β oxidation in the liver.

Re-discussion on the comprehensive treatment strategy of complex ventral hernia from the perspective of intraperitoneal pressure / 中华外科杂志

Complex ventral hernia refers to a large hernia that is complicated by a series of concurrent conditions. Change in intra-abdominal pressure is one of the main pathways through which various factors exert an impact on perioperative risk and postoperative recurrence. Taking abdominal pressure reconstruction as the core, the treatment strategy for complex abdominal hernia can be formulated from three aspects: improving patients' tolerance, expanding abdominal cavity volume, and reducing the volume of abdominal contents. Improving patients' tolerance includes abdominal wall compliance training and progressive preoperative pneumoperitoneum. To expand the volume of the abdominal cavity, implanting hernia repair materials, component separation technique, autologous tissue transplantation, component expend technique, and chemical component separation can be used. Initiative content reduction surgery and temporary abdominal closure may be performed to reduce the volume of abdominal contents. For different cases of complex ventral hernia, personalized treatment measures can be safely and feasibly adopted depending on the condition of the patients and the intra-abdominal pressure situation.

A comparative study of the clinical characteristics of spotted fever and severe fever with thrombocytopenia syndrome / 中国热带医学

@#Abstract: Objective To investigate the differences in epidemiological and clinical characteristics of patients with spotted fever (SF) and severe fever with thrombocytopenia syndrome (SFTS). Methods A total of 86 patients with SF and 113 patients with SFTS who were laboratory-confirmed in the second-level and above hospitals in Lu'an City from January 2017 to January 2022 were selected. The basic data, epidemiological history, clinical data and laboratory test results of the two diseases were retrospectively analyzed for comparison. Results The proportion of male in SF group was 32.56% (28/86), and the proportion of male in SFTS group was 53.98% (61/113), the difference was statistically significant (χ2=9.067, P<0.01). The proportions of abdominal pain and diarrhea in the SF group were (3.49%, 3/83) and (21.24%, 24/113), which were significantly lower than corresponding (6.98%, 6/86) and (46.90%, 53/113) in the SFTS group (χ2=13.121, 37.322, P<0.01). The incidences of rash and eschar in SF group were 95.35% (82/86) and 20.93% (18/86), which were significantly higher than corresponding 1.77% (2/113) and 0.88% (1/113) in SFTS group (χ2=175.311, 22.721, P<0.01). The levels of leukocytes, platelets and C-reactive protein in the SF group were significantly higher than those in the SFTS group, and the levels of transaminase, lactate dehydrogenase and D-dimer were significantly lower than those in the SFTS group, and the differences were statistically significant (all P<0.05). Conclusions The rash and inflammatory reaction are more obvious in SF patients, while the liver function, myocardial function and coagulation function are significantly impaired in SFTS patients.

Clinical characteristics and prognosis of primary and secondary diffuse large B-cell lymphoma of the pancreas / 中华血液学杂志

Objective: To analyze the clinical characteristics and prognosis of primary and secondary pancreatic diffuse large B-cell lymphoma (DLBCL) . Methods: Clinical data of patients with pancreatic DLBCL admitted at Shanghai Rui Jin Hospital affiliated with Shanghai Jiao Tong University School of Medicine from April 2003 to June 2020 were analyzed. Gene mutation profiles were evaluated by targeted sequencing (55 lymphoma-related genes). Univariate and multivariate Cox regression models were used to evaluate the prognostic factors of overall survival (OS) and progression-free survival (PFS) . Results: Overall, 80 patients were included; 12 patients had primary pancreatic DLBCL (PPDLBCL), and 68 patients had secondary pancreatic DLBCL (SPDLBCL). Compared with those with PPDLBCL, patients with SPDLBCL had a higher number of affected extranodal sites (P<0.001) and had higher IPI scores (P=0.013). There was no significant difference in the OS (P=0.120) and PFS (P=0.067) between the two groups. Multivariate analysis indicated that IPI intermediate-high/high risk (P=0.025) and double expressor (DE) (P=0.017) were independent adverse prognostic factors of OS in patients with pancreatic DLBCL. IPI intermediate-high/high risk (P=0.021) was an independent adverse prognostic factor of PFS in patients with pancreatic DLBCL. Targeted sequencing of 29 patients showed that the mutation frequency of PIM1, SGK1, BTG2, FAS, MYC, and MYD88 in patients with pancreatic DLBCL were all >20%. PIM1 (P=0.006 for OS, P=0.032 for PFS) and MYD88 (P=0.001 for OS, P=0.017 for PFS) mutations were associated with poor OS and PFS in patients with SPDLBCL. Conclusion: There was no significant difference in the OS and PFS between patients with PPDLBCL and those with SPDLBCL. IPI intermediate-high/high risk and DE were adverse prognostic factors of pancreatic DLBCL. PIM1, SGK1, BTG2, FAS, MYC, and MYD88 were common mutations in pancreatic DLBCL. PIM1 and MYD88 mutations indicated worse prognosis.

Detection and analysis of neutralizing antibodies against SARS-CoV-2 in convalescent plasma donors / 中国输血杂志

【Objective】 To detect the anti-SARS-CoV-2 neutralizing antibody levels in convalescent plasma (CP) and to evaluate whether it has specific anti-SARS-CoV-2 S antigen effect, so as to provide laboratory data support for clinical use of CP. 【Methods】 Nine CP donors who have recovered from COVID-19 were studied, and 4 volunteers who completed the vaccination and 3 asymptomatic infected blood donors were compared. Anti-SARS-CoV-2 antibodies including total antibody, IgM and IgG were measured by chemiluminescence microparticle immunoassays (CMIA) test in three groups. The VSV pseudovirus-based neutralization assay for evaluating neutralizing antibodies against SARS-CoV-2 was carried out in all samples. 【Results】 All samples were tested positive by the total antibody and IgG CMIA in COVID-19 CP donors and recipients of inactivated COVID-19 vaccine. High titers of IgG were observed in CP donors and vaccine recipients compared with asymptomatic blood donors. All vaccine recipients and 8 of 9 CP donors tested positive by SARS-CoV-2 pseudovirus-based neutralization test, whereas all asymptomatic blood donors tested negative. 【Conclusion】 The levels and characteristics of neutralizing antibodies among COVID-19 CP donors, vaccine recipients and asymptomatic blood donors were different. When unable to implement the pseudovirus assay to measure neutralizing antibodies, the detection of total antibody can be considered instead.

Determination of methylene blue release in plasma virus inactivation bag by fluorescence energy transfer method / 中国输血杂志

【Objective】 To establish a simple, economical and rapid method for the determination of methylene blue (MB) release in virus inactivation bag. 【Methods】 Based on the fluorescence energy transfer between MB and BSA-stabilized gold nanoclusters (BSA-AuNCs), the standard curve of MB determination was established by measuring the fluorescence quenching degree of MB to BSA-AuNCs in different concentrations to conduct the determination of MB release in virus inactivation bag. 【Results】 There was a good linear relationship between the MB concentration (cMB) and the fluorescence quenching degree of BSA-AuNCs[ (I0-I)/I0=0.018cMB+ 0.021(r=0.996)] when the fluorescence emission wavelength was about 620 nm and the cMB was in the range of (0.9-36) μmoL/L. The recovery of MB was 98.00% -101.95 % when applied to determine MB at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.73%, 0.81% and 0.77% respectively, and the obtained inter-day variation coefficients were 3.92%, 3.81%, and 4.73% respectively. There was no significant difference between the results measured by this method and those measured by combination of solid-phase extraction and spectrophotometry(P>0.05). 【Conclusion】 The fluorescence energy transfer method could achieve simple and rapid determination of MB release in virus inactivation bag with accurate and reliable results.

Determination of fibrinogen content in cryoprecipitated antihemophilic factor by fluorescence quenching method / 中国输血杂志

【Objective】 To establish a new method for the determination of fibrinogen content in cryoprecipitated antihemophilic factor. 【Methods】 Fibrinogen (Fib) could bind with sheep anti-human fibrinogen (anti-Fib) specifically and further form antigen-antibody complex. When the Fib was present in the solution, the fluorescence of fluorescein isothiocyanate (FITC) labeled on the anti-Fib (FITC-anti-Fib) was quenched due to the formation of immune complex. The fluorescence quenching degree of FITC-anti-Fib was positively correlated with Fib concentration (cFib) in a certain concentration range. 【Results】 The linear relationship between fluorescence quenching degree [(I0-I)/I0] of FITC-anti-Fib and ln(cFib) was (I0-I)/I0=15.53ln(cFib)+ 80.79 (R2=0.99) when the cFib was in the range of (0.007 8-0.560 0) g/L. The recovery of Fib was (96.77-102.43) %. When the method was applied to determine Fib at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.31%, 0.56%, and 0.49%, respectively, and the inter-day variation coefficients were 3.81%, 3.06%, and 4.13%, respectively. There was no significant difference between the results measured by fluorescence quenching method and coagulation method (t=-0.075, P>0.05). 【Conclusion】 In this work, a new fluorescence method for the determination of Fib in cryoprecipitated antihemophilic factor was successfully established based on the specific combination of fib and FITC-anti-Fib. The method is simple and rapid. The obtained results were accurate and reliable by using this method to determine Fib.

Screening Novel Drugresistant Mutations in Mycobacterium tuberculosis Strains by Using the Amplicon Sequencing Method / 中国生物化学与分子生物学报

Tuberculosis is one of the seriously public problems. The increasing drug-resistant tuberculosis is the key problem for controlling tuberculosis. Rapid and accurate diagnosis is important for further treatment. In this study‚ a next-generation sequencing method based on amplicon sequencing was constructed to screen the mutations in 17 drug-resistant genes of five first-line anti-tuberculosis drugs. A total of 65 mutations were identified in 26 clinic drug-resistant tuberculosis strains‚ including 33 hotspot mutations‚ 9 rare mutations‚ and 23 novel mutations. The pathogenesis‚ conservation‚ and partial structures caused by 18 novel missense mutations were predicted. The results showed that 14 novel mutations showed high conservation in nine species. All these 14 mutations could change the partial structure of protein. According to the detection and analysis results of this study‚ it is speculated that these newly discovered mutations may be potential drug-resistant mutations. It is a rapid‚ accurate and comprehensive method for the detection of drug-resistant mutations in first-line drug-resistant Mycobacterium tuberculosis‚ which could identify hotspot and rare mutations together with novel mutations. The detection method may be used for clinical diagnosis and basic research.

The protective effect of fluoxetine hydrochloride on brain tissues of rats with high altitude cerebral edema / 中国药理学通报

Aim To study the protective effect of fluoxetine hydrochloride on brain tissues of rats with simulated high altitude cerebral edema(HACE)and its mechanism.Methods The optimal dosage and time of fluoxetine hydrochloride were determined by the hypoxia tolerance test of mice under normal pressure.The rat model of brain edema at high altitude was established by large-scale low-pressure oxygen chamber.HE staining was used to observe the pathological changes of brain tissues in rats.Microplate reader was used to detect the corresponding indexes of oxidative stress such as malondialdehyde(MDA)level and superoxide dismutase(SOD)activity.The expressions of hypoxia-related proteins HIF-1α,VEGF,MMP-9,AQP4 and SERT were detected by Western blot.Results Compared with the hypoxia model group,after the intervention of fluoxetine hydrochloride,the survival time of mice was prolonged,and the middle dose of fluoxetine(14 mg·kg-1)had the best effect,with an extension rate of 17.78%.The pathological damage of brain was improved,the water content of brain decreased,and the permeability of blood-brain barrier decreased.MDA content in rat brain decreased and SOD activity increased.Western blot results showed that HIF-1α,VEGF,MMP-9,AQP4,SERT protein were significantly down-regulated.Conclusions Fluoxetine has protective effect on rats with brain edema at high altitude,and its mechanism may be related to improving oxidative stress,activating HIF-1α/VEGF/MMP-9 signaling pathway and affecting the expression of SERT protein.SERT may be a potential target for treating brain edema at high altitude.

Study on the changes of cellular immune function after endoscopic radiofrequency ablation in extrahepatic cholangiocarcinoma / 肿瘤研究与临床

Objective:To investigate the effect of radiofrequency ablation treatment of extrahepatic cholangiocarcinoma under endoscopic retrograde cholangiopancreatography on the cellular immune function of patients with extrahepatic cholangiocarcinoma.Methods:A prospective cohort study was conducted on 78 patients with inoperable extrahepatic cholangiocarcinoma who visited Shanxi Provincial People's Hospital from October 2019 to January 2022. All of 78 patients underwent endoscopic radiofrequency ablation. Peripheral venous blood was collected from all patients at 1 day before treatment, 1 week after treatment and 1 month after treatment. Lymphocyte subsets were detected by flow cytometry, and levels of cytokines were detected by enzyme-linked immunosorbent assay. Seventy-eight healthy subjects with the same age and gender as the patients were selected as healthy controls, and the same indicators were tested.Results:The patient's peripheral blood total bilirubin was (193±83) μmol/L at 1 day before treatment, and decreased to (55±49) μmol/L at 1 week after treatment ( t = 16.70, P < 0.001); γ-glutamyltransferase was (242±108) U/L at 1 day before treatment, and decreased to (92±70) U/L at 1 week after treatment ( t = 19.12, P < 0.001). The proportion of natural killer (NK) cells, CD3 + T cells, CD4 + T cells, ratio of CD4 + T cells to CD8 + T cells (CD4/CD8) and level of interferon γ (IFN-γ) of patients at 1 day before treatment were lower than those of healthy control group (all P < 0.05), the proportion of CD8 + T cells and regulatory T cells was higher than those of healthy control group (both P < 0.001), and there was no significant difference in the levels of interleukin 4 (IL-4) between the patients at 1 day before treatment and the healthy control group ( P = 0.545). At 1 week and 1 month after treatment, the proportion of NK cells, CD3 + T cells, CD4 + T cells, CD4/CD8 and IFN-γ level in patients increased compared with 1 day before treatment (all P < 0.001), and decreased at 1 month after treatment compared with 1 week after treatment (all P < 0.001). At 1 week and 1 month after treatment, the proportion of CD8 + T cells and IL-4 level in patients decreased compared with 1 day before treatment (both P < 0.001), and increased at 1 month after treatment compared with 1 week after treatment (both P < 0.001). At 1 week and 1 month after treatment, the proportion of regulatory T cells decreased compared with 1 day before treatment (both P < 0.001), and decreased at 1 month after treatment compared with 1 week after treatment ( P < 0.001). Conclusions:Patients with extrahepatic cholangiocarcinoma are at a low level of immunity. Endoscopic radiofrequency ablation treatment can alleviate the patient's immunosuppression, enhance the patient's cellular immunity.

Prognostic factors of grade Ⅲ- Ⅳ mixed hemorrhoids with anorectal resting hypertension and the therapeutic effect of MMH / 国际外科学杂志

Objective:To investigate the effect of external dissection and internal ligation (MMH) in the treatment of grade Ⅲ to Ⅳ mixed hemorrhoids with anorectal resting hypertension and its prognostic risk factors.Methods:The clinical data of 48 patients with grade Ⅲ-Ⅳ mixed hemorrhoids complicated with rectal and anal resting hypertension treated by MMH in Nanjing Jiangbei people′s Hospital Affiliated to Nantong University from February 2018 to February 2020 were analyzed retrospectively (observation group), including 24 males and 24 females; The age ranged from 22 to 55 (41.87±7.52) years. Another 48 patients with grade Ⅲ-Ⅳ mixed hemorrhoids complicated with anorectal resting hypertension treated by PPH were selected as the control group, including 20 males and 28 females; The average age was (42.68±7.14) years. The clinical effective rates , pain score at 6, 24 and 72 h after operation, postoperative complications were compared between the two groups. Multiple logistic regression was used to analyze the related risk factors affecting the prognosis.Results:There was no significant difference in VAS score between the two groups at 6 h after operation ( t=0.25, P=0.807); the VAS score of the observation group at 24 h and 72 h after operation was lower than that of the control group ( t=7.044, P<0.001; t=5.307, P<0.001); the total effective rate of the observation group was higher than that of the control group (93.75% vs 77.08%, χ2=5.35, P=0.021); the total incidence of postoperative anal edema, defecation difficulty, bloody stool, anal pain and fecal incontinence in the observation group was lower than that in the control group, the difference was statistically significant (12.50% vs 33.33%, χ2=5.879, P=0.015). Age, course of disease, grading of internal hemorrhoids and treatment methods were related to the prognosis of patients ( P<0.05); logistic regression analysis showed that age (> 45 years), course of disease (>10 years), grade of internal hemorrhoids (grade Ⅳ) and treatment (PPH) were independent risk factors for prognosis of patients ( P<0.05). Conclusions:MMH has less complications, less postoperative pain and satisfactory curative effect in the treatment of Ⅲ to Ⅳ mixed hemorrhoids and anorectal resting hypertension. The older the patients, the longer the course of disease, the higher the degree of internal hemorrhoids and the choice of treatment methods were the risk factors for the prognosis. Early diagnosis and reasonable treatment are helpful to improve the prognosis of patients.

Inhibition of Ciliogenesis Enhances the Cellular Sensitivity to Temozolomide and Ionizing Radiation in Human Glioblastoma Cells / 生物医学与环境科学（英文）

Objective@#To investigate the function of primary cilia in regulating the cellular response to temozolomide (TMZ) and ionizing radiation (IR) in glioblastoma (GBM).@*Methods@#GBM cells were treated with TMZ or X-ray/carbon ion. The primary cilia were examined by immunostaining with Arl13b and γ-tubulin, and the cellular resistance ability was measured by cell viability assay or survival fraction assay. Combining with cilia ablation by IFT88 depletion or chloral hydrate and induction by lithium chloride, the autophagy was measured by acridine orange staining assay. The DNA damage repair ability was estimated by the kinetic curve of γH2AX foci, and the DNA-dependent protein kinase (DNA-PK) activation was detected by immunostaining assay.@*Results@#Primary cilia were frequently preserved in GBM, and the induction of ciliogenesis decreased cell proliferation. TMZ and IR promoted ciliogenesis in dose- and time-dependent manners, and the suppression of ciliogenesis significantly enhanced the cellular sensitivity to TMZ and IR. The inhibition of ciliogenesis elevated the lethal effects of TMZ and IR via the impairment of autophagy and DNA damage repair. The interference of ciliogenesis reduced DNA-PK activation, and the knockdown of DNA-PK led to cilium formation and elongation.@*Conclusion@#Primary cilia play a vital role in regulating the cellular sensitivity to TMZ and IR in GBM cells through mediating autophagy and DNA damage repair.