Objective:To analyze the correlation between variants in the start codon of the α-globin gene and phenotypes of thalassemia, so as to provide a basis for the diagnosis and prevention of α-thalassemia.Methods:A retrospective study was conducted on 7 patients diagnosed by Yangjiang People′s Hospital and Guangzhou Hybribio Co. Ltd., from June 2019 to October 2022. Routine blood tests and hemoglobin electrophoresis were carried out. Potential variants were identified through polymerase chain reaction (PCR) combined with Reverse dot blotting (RDB), Gap-PCR, and Sanger sequencing. This study has been approved by the Medical Ethics Committee of People′s Hospital of Yangjiang (Ethics No: 20240001).Results:For the 7 patients, results of blood routine test of one case was unknown, and that of another was normal. The remaining 5 cases had presented with microcytic hypochromic anemia. The results of hemoglobin electrophoresis showed that one case had normal Hb A and slightly lower Hb A 2, whilst another had significantly decreased Hb A and Hb A 2, in addition with the appearance of a Hb H band. The content of Hb Bart′s in four neonates was ≥0.4%. The remaining one case had no result. Genetic testing has identified 4 rare start codon mutations, namely HBA2: c. 2delT, HBA2: c. 1A>G, HBA2: c. 1A>T, and HBA1: c. 2T>C. Among these, Patient 1 had harbored compound heterozygous variants of HBA2: c. 427T>C (Hb CS) and HBA2: c. 2delT. Patient 4 had harbored compound heterozygous variants of HBA2: c. 1A>G and Southeast Asian type deletion. Conclusion:Heterozygotes with HBA start codon variants usually present as silent or mild thalassemia, and the symptoms of anemia may deteriorate when combined with other α-thalassemia variant. The HBA2: c. 1A>T start codon variant was unreported previously in China. The detection of start codon variants has helped to clarify the causes of anemia, genetic counseling, and guidance for reproduction.

OBJECTIVE: To analyze the correlation between variants in the start codon of the α-globin gene and phenotypes of thalassemia, so as to provide a basis for the diagnosis and prevention of α-thalassemia. METHODS: A retrospective study was conducted on 7 patients diagnosed by Yangjiang People's Hospital and Guangzhou Hybribio Co. Ltd., from June 2019 to October 2022. Routine blood tests and hemoglobin electrophoresis were carried out. Potential variants were identified through polymerase chain reaction (PCR) combined with Reverse dot blotting (RDB), Gap-PCR, and Sanger sequencing. This study has been approved by the Medical Ethics Committee of People's Hospital of Yangjiang (Ethics No: 20240001). RESULTS: For the 7 patients, results of blood routine test of one case was unknown, and that of another was normal. The remaining 5 cases had presented with microcytic hypochromic anemia. The results of hemoglobin electrophoresis showed that one case had normal Hb A and slightly lower Hb A2, whilst another had significantly decreased Hb A and Hb A2, in addition with the appearance of a Hb H band. The content of Hb Bart's in four neonates was ≥ 0.4%. The remaining one case had no result. Genetic testing has identified 4 rare start codon mutations, namely HBA2: c.2delT, HBA2: c.1A>G, HBA2: c.1A>T, and HBA1: c.2T>C. Among these, Patient 1 had harbored compound heterozygous variants of HBA2: c.427T>C (Hb CS) and HBA2: c.2delT. Patient 4 had harbored compound heterozygous variants of HBA2: c.1A>G and Southeast Asian type deletion. CONCLUSION Heterozygotes with HBA start codon variants usually present as silent or mild thalassemia, and the symptoms of anemia may deteriorate when combined with other α-thalassemia variant. The HBA2: c.1A>T start codon variant was unreported previously in China. The detection of start codon variants has helped to clarify the causes of anemia, genetic counseling, and guidance for reproduction.
Humans ; Phenotype ; Codon, Initiator/genetics* ; Female ; Male ; Retrospective Studies ; alpha-Globins/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobin A/genetics* ; Adult ; Mutation

Objective:To analyze the correlation between variants in the start codon of the α-globin gene and phenotypes of thalassemia, so as to provide a basis for the diagnosis and prevention of α-thalassemia.Methods:A retrospective study was conducted on 7 patients diagnosed by Yangjiang People′s Hospital and Guangzhou Hybribio Co. Ltd., from June 2019 to October 2022. Routine blood tests and hemoglobin electrophoresis were carried out. Potential variants were identified through polymerase chain reaction (PCR) combined with Reverse dot blotting (RDB), Gap-PCR, and Sanger sequencing. This study has been approved by the Medical Ethics Committee of People′s Hospital of Yangjiang (Ethics No: 20240001).Results:For the 7 patients, results of blood routine test of one case was unknown, and that of another was normal. The remaining 5 cases had presented with microcytic hypochromic anemia. The results of hemoglobin electrophoresis showed that one case had normal Hb A and slightly lower Hb A 2, whilst another had significantly decreased Hb A and Hb A 2, in addition with the appearance of a Hb H band. The content of Hb Bart′s in four neonates was ≥0.4%. The remaining one case had no result. Genetic testing has identified 4 rare start codon mutations, namely HBA2: c. 2delT, HBA2: c. 1A>G, HBA2: c. 1A>T, and HBA1: c. 2T>C. Among these, Patient 1 had harbored compound heterozygous variants of HBA2: c. 427T>C (Hb CS) and HBA2: c. 2delT. Patient 4 had harbored compound heterozygous variants of HBA2: c. 1A>G and Southeast Asian type deletion. Conclusion:Heterozygotes with HBA start codon variants usually present as silent or mild thalassemia, and the symptoms of anemia may deteriorate when combined with other α-thalassemia variant. The HBA2: c. 1A>T start codon variant was unreported previously in China. The detection of start codon variants has helped to clarify the causes of anemia, genetic counseling, and guidance for reproduction.

OBJECTIVE To investigate the effect of Guanxinning tablets(GXN) on early heart failure model rats, and to explore the protective mechanism of GXN on heart failure rats from the perspective of intestinal flora. METHODS Six rats who underwent sham operation were set as sham operation group. Took 80 SD rats to undergo aortic arch stenosis and established a heart failure rat model. The surviving rats were divided into 4 groups, namely the model control group, the positive control group(captopril tablets 12.5 mg·kg–1), high-dose and low-dose of GXN group(600, 1 200 mg·kg–1). The 4 groups were administered continuously for 8 weeks. Cardiac ultrasonography was performed every 4 week. Serum NT-proBNP, hs-CRP, IL-6, TNF-α, SOD and MDA levels were measured. The effects of GXN on the structure and function of intestinal flora were observed based on the high-throughput sequencing technology and bioinformatics analysis of 16S gut microbiome. RESULTS Compared to the model control group, after giving different doses of GXN, the survival rate of rats increased, and the thickness of the ventricular wall decreased to varying degrees. The weight of the heart and coefficient of the heart were all reduced. GXN could also reduce the level of inflammatory factors, inhibit the level increase of NT-proBNP in rats, and increase the activity of serum SOD. In addition, GXN intervention could significantly improve the intestinal flora diversity of rats with heart failure, the possible target genera of GXN were Akkermansia genera, Phascolarctobacterium genera and Oxalobacter genera. The effect of GXN on intestinal function in rats with heart failure might be concentrated in non-homologous end-joining, influenza A, carotenoid synthesis, indole alkaloids biosynthesis, betalain biosynthesis, renin-angiotensin system and other biological pathways. CONCLUSION The protective effect of GXN on early heart failure rats may be related to the regulation of intestinal flora pathway.

Objective:To explore the hematological phenotype and genotypic characteristics of five Chinese individuals with a rare thalassemia mutation HBB: c. 93-21G>A. Methods:A retrospective study was carried out on five individuals identified by the People′s Hospital of Yangjiang and Guangzhou Hybribio Co., Ltd. from May 2018 to September 2022. Routine blood test and hemoglobin electrophoresis were performed, and the genotypes of five subjects were determined by using PCR combined with reverse dot blotting (RDB), nested PCR, Gap-PCR and Sanger sequencing. This study was approved by Medical Ethics Cornmittee of the People′s Hospital of Yangjiang (Ethics No. 20240001).Results:Among the five individuals, hematological data of one was unavailable, and the remaining four had presented with microcytosis and hypochromia. The results of hemoglobin electrophoresis indicated that all of them had a HbA 2 level of ≥4.7%. Genetic analysis showed that one case had harbored compound heterozygous mutations of ααα anti3.7 triplet and HBB: c. 93-21G>A, one had compound heterozygous mutations of -α 3.7 and HBB: c. 93-21G>A, whilst the remaining three were heterozygous for the HBB: c. 93-21G>A mutation. Conclusion:The hematological phenotype of β-thalassemia carriers ( HBB: c. 93-21G>A) is similar to that of other β + thalassemia heterozygotes with mild β-thalassemia characteristics.

OBJECTIVE：Compare the fingerprint difference of Vaccariae Semen before and after processed （stir-fried），and to determine the contents of erythrine and vaccarin before and after stir-fried. METHODS ：UPLC method was adopted. The determination was performed on YMC Trait C 18 column with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 219 nm，and the column temperature was 35 ℃. The sample size was 1 μL. Using vaccarin as reference，the fingerprints of Vaccariae Semen crude product and its processed product （each of 17 batches，S1-S17，CS18-CS34） were drawn. The similarity evaluation and common peak identification were carried out by Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）；cluster analysis ，principle component analysis （PCA）and factor analysis were performed by using SPSS 20.0 software. The contents of erythrine and vaccarin in Vaccariae Semen crude product and its processed product were determined by UPLC. RESULTS ：There were 5 common peaks in UPLC fingerprints of 17 batches of Vaccariae Semen crude product and its processed product. The similarities were all higher than 0.99. Among them ， 2 common peaks were identified ，i.e. erythrine ，vaccarin. Results of cluster analysis showed that S 1-S17 were clustered into one category and CS 18-CS34 were clustered into one category. Results of PCA and factor analysis showed that variance contribution rate of the first principle component was 76.418％；erythrine and vaccarin had higher loading on the first principal component （eigenvalues were 0.976 and 0.966，respectively）. The linear ranges of above 2 components were 6.437-321.832 μg/mL and 7.729-386.437 μg/mL，respectively（r＞0.999）. The limits of detection and quantitation were 0.085，0.284 ng （crude product） and 0.739， 2.465 ng （processed product ）， respectively. RSDs of precision ，reproducibility，stability（12 h）and durability tests were all lower than 3％（n＝6 or n＝5）. E-mail：1083656123@qq.com Average recoveries were 96.42％（RSD＝0.85％，n＝6）and 99.13％（RSD＝1.74％，n＝6）. The contents of the two components were 0.11％-0.20％，0.42％-0.63％（crude product ）and 0.08％-0.11％，0.34％-0.50％（processed product ）. CONCLUSIONS ：UPLC fingerprint of Vaccariae Semen crude product and its processed product are established successfully. Although the chemical constituents in Vaccariae Semen are consistent before and after stir-fried ，the contents of erythrine and vaccarin are all decreased after stir-fried.

Objective To investigate the detection and distribution of hospitalized specimens from a tertiary hospital over 5 years. Methods Specimens of sputum, urine, blood, secretions and puncture fluid were collected from patients admitted to the Harrison International Peace Hospital from November 2013 to November 2018. The origin of specimens, the distribution of departments and the distribution of pathogenic bacteria isolated were analyzed retrospectively. Results A total of 61 286 specimens were sent for examination during the 5 years. The top 5 specimens were sputum culture (n = 18 302, 29.9%), sputum smear (n = 11 253, 18.4%), blood culture (n = 9 713, 15.8%), urine culture (n = 6 448, 10.5%) and secretion culture (n = 6 133, 10.0%), accounting for 84.6% (51 849/61 286). Sputum specimens accounted for 48.2% (29 555/61 286) with the largest proportion. The number of specimens from medical wards was much higher than that from surgical wards (specimens: 25 468 vs. 10 521), respiratory medicine, department of critical care medicine and emergency intensive care unit (EICU) were important sources of pathogenic specimens in the hospital, accounting for 29.8% (18 243/61 286) in total. The average positive rate of all specimens was 23.5% (14 424/61 286). The positive rates of sputum culture and urine culture were 29.7% (5 428/18 302) and 35.4% (2 281/6 448), respectively, while the positive rate of blood culture was only 6.6% (643/9 713). Escherichia coli was the most common pathogen in all specimens except for sputum culture and fecal culture. Escherichia coli [40.6% (926/2 281)], Klebsiella pneumoniae [9.2% (210/2 281)], Pseudomonas aeruginosa [8.2% (188/2 281)], Enterococcus faecalis (group D) [6.6% (151/2 281)] and Candida albicans [3.2% (73/2 281)] were the most common pathogens in urine culture. Klebsiella pneumoniae [24.1% (1 309/5 428)], Acinetobacter baumannii [21.3% (1 154/5 428)], Pseudomonas aeruginosa [15.1% (818/5 428)], Escherichia coli [6.5% (351/5 428)] and Maltose oligotrophomonas maltose [5.8% (316/5 428)] were the most common pathogens in sputum culture. Escherichia coli [36.5% (235/643)], Klebsiella pneumoniae [10.9% (70/643)], Pseudomonas aeruginosa [4.8% (31/643)], Staphylococcus epidermidis [3.4% (22/643)] and Staphylococcus humanis [3.3% (21/643)] were the most common pathogens in blood culture. Conclusion Specimens sent for examination by inpatients are mainly from internal medicine wards, mainly from sputum, blood and urine, and the detected pathogens are mainly Gram-negative bacteria.

BACKGROUND: Mesenchymal stem cells in Wharton's Jelly of the human umbilical cord can induce differentiation into islet-like cells.OBJECTIVE: To verify the possibility of human umbilical cord derived mesenchymal stem cells co-cultured with rat pancreatic cells differentiate into islet-like cells, and to observe the effects of transplantation of islet-like cells on blood glucose of diabetic rats.METHODS: Mesenchymal stem cells in Wharton's Jelly of the human umbilical cord was separated, induced, passaged, and co-cultured with pancreatic cells to induce differentiation into islet-like clusters. Rats were divided into the normal control, model and experimental groups. Rats in the model group were prepared for diabetic models, and those in the experimental group were transplanted islet-like cells after model preparation. RESULTS AND CONCLUSION: There were cells crawled out of cultured Wharton's Jelly of the human umbilical cord, and morphology of adhered cells turned into fusiform shape at 7 days. The isolated cells are characterized by expressing specific surface markers of mesenchymal stem cells, such as CD44, CD29, CD105, but not expressing CD34, CD45 or CD14. The cells were strongly stained by PDX-1 and human insulin at 7 and 10 days. Compared with the simple culture group, the expression of human insulin and concentration of C-peptide were obviously increased; PDX-1 and human insulin mRNA expressions were highly expressed at 7 and 10 days after induction. Compared with the model group, the streptozotocin test of rats in the experimental group was obvious decreased (P < 0.01), but extremely higher than that of the normal control group at 1 week after transplantation (P < 0.01). Brdu positive nuclei and insulin positive kytoplasms could be seen in the experimental group at 8 weeks after transplantation. The results demonstrated that, umbilical cord derived mesenchymal stem cells existed in Wharton's Jelly. The co-cultured cells promote mesenchymal stem cells differentiating into islet-like cells, which can dramatically decrease blood glucose in diabetic rats.

A novel method based on pH value was proposed to simplify the substrate feeding method for glutamic acid fermentation. The linear relationship between the consumption amounts of ammonia (x) and that of glucose (y) was established (y = 7.4744x, R2 = 0.9989) which could be used as the ratio of the amount of ammonia and that of glucose in the feeding broth. Thus the concentration of glucose could be controlled through the adjustment of pH automatically. In the glutamic acid fermentation using the pH feedback-controlled glucose feeding method, the glucose concentration in fermentation broth was maintained between 12 and 21 g/L. Compare with the constant glucose concentration feeding method, the glucose conversion rate and glutamic acid productivity increased by 9.06% and 17.5% respectively, when the pH feedback-controlled glucose feeding method was employed, and fermentation period was shorten above 2 h.
Bioreactors ; microbiology ; Corynebacterium glutamicum ; growth & development ; metabolism ; Culture Techniques ; Feedback ; Fermentation ; Glucose ; metabolism ; Glutamic Acid ; biosynthesis ; metabolism ; Hydrogen-Ion Concentration ; Quaternary Ammonium Compounds ; metabolism

Endotoxin removal is essential for the safety of biological products. To remove endotoxin efficiently, we used polymyxin B (PMB) affinity adsorbent to remove endotoxin from protein solutions by static adsorption. We studied the effects of spacer length and ligand density of the affinity adsorbent, pH, salt type and concentration, protein type and concentration, endotoxin concentration, and additive on endotoxin removal and protein recovery. Endotoxin content and protein concentration were determined by test and Lowry assay respectively. The results showed that PMB affinity adsorbent had high capacity, high adsorption speed, high removal efficiency and good reusability. In addition, ligand density, pH, salt concentration and the isoelectric point and hydrophobicity of protein all had remarkable influence on the endotoxin removal. Under the optimal conditions, the recoveries of hemoglobin, human serum albumin and lysozyme were 87.2%, 73.4% and 97.3%, respectively, and the corresponding endotoxin removal rates 99.8%, 97.9% and 99.7%, respectively. This study illustrated the effects of solution conditions on the efficiency of endotoxin removal and protein recovery, and would provide useful reference for the efficient removal of endotoxin from biological products.
Adsorption ; Chromatography, Affinity ; methods ; Drug Contamination ; prevention & control ; Endotoxins ; isolation & purification ; Hemoglobins ; isolation & purification ; Polymyxin B ; chemistry ; Proteins ; isolation & purification ; Pyrogens ; isolation & purification ; Serum Albumin ; isolation & purification ; Solutions