Genotype and Phenotype of α-Thalassemia Fusion Gene in Huadu District of Guangzhou, Guangdong Province of China / 中国实验血液学杂志

OBJECTIVE@#To explore the carrier rate, genotype and phenotype of α-thalassemia fusion gene in Huadu district of Guangzhou, Guangdong province of China, and provide data reference for the prevention and control of thalassemia.@*METHODS@#A total of 10 769 samples who were screened for thalassemia in Maternal and Child Health Hospital of Huadu District from July 2019 to November 2020 were analyzed retrospectively. Blood cell analysis and hemoglobin (Hb) electrophoresis were performed. Thalassemia genes were analyzed by gap-PCR and PCR-reverse dot blot hybridization (PCR-RDB).@*RESULTS@#A total of 9 cases with α-thalassemia fusion gene were detected in 10 769 samples (0.08%). There were 7 cases with fusion gene heterozygote, 1 case with compound of α-thalassemia fusion gene and Hb G-Honolulu, 1 case with compound of α-thalassemia fusion gene and Hb QS. The MCV results of 4 samples of blood cell analysis were within the reference range, the Hb A2 value of 1 case was decreased, and there were no other abnormalities found.@*CONCLUSION@#The α-thalassemia fusion gene is common in Huadu district of Guangzhou, and heterozygotes are more common, and current screening methods easily lead to misdiagnosis.

The Recent HIV-1 Infections in Blood Donors of Fuzhou Zone / 中国实验血液学杂志

OBJECTIVE@#To investigate the recent HIV-1 infections of the blood donors in Fuzhou zone.@*METHODS@#The positive HIV antibody confirmatory samples in Fuzhou zone from 2012 to 2016 were collected and tested by LAg-Avidity EIA, and HIV long-term infections or recent infections were determined.@*RESULTS@#405 371 cases of blood donors were tested in the period from 2012 to 2016, and 94 HIV confirmatory positive samples were collected. 35 cases were recent infections determined by LAg-Avidity EIA, the annual HIV-1 incidences were 1.326‰, 0.845‰, 0.694‰, 1.148‰ and 0.364‰, the average incidences were 0.863‰. Among 94 cases of HIV confirmatory positive donors，58 cases were first donors and 36 cases were repeated donors, 17(29.3%) and 18 (50.0%) cases were recent infections respectively, which showed statistical significance（χ@*CONCLUSION@#The HIV-1 incidences were stable among blood donors in Fuzhou zone. The percentage of HIV-1 recent infections in repeated donors were more higher than that in first donors.

Correlations between ultrasonographic and subsequent radiographic findings of developmental dysplasia of the hips

Purpose@#The study aimed to investigate the utility of ultrasonographic (US) findings in predicting the subsequent radiographic parameters of developmental dysplasia of the hips. @*Methods@#In this 12-year retrospective cohort study, all new-born infants with a positive clinical examination or risk factors were included. They were scheduled for hip ultrasonography in the first 3 months, and subsequent radiographs at 1 year of life. The US images were evaluated using the Graf classification, Harcke’s dynamic screening method, and Terjesen’s femoral head coverage method. The radiographic images were evaluated using the acetabular index and femoral head position. The overall US or radiographic findings were considered abnormal if they were classified as abnormal for any of their respective parameters. The overall US and radiographic parameters were correlated. @*Results@#A total of 160 patients were included. The overall US and radiographic parameters showed no statistically significant difference (P=0.050). The sensitivity, specificity, and accuracy of the overall US parameters were 57.1%, 84.9%, and 81.3%, respectively. All three individual US parameters showed no statistically significant differences, with the overall radiographic findings and acetabular index (P>0.05). However, they showed a statistically significant difference, with the position of the femoral head (P<0.001), with the US parameters having an excellent negative predictive value of 100% for identifying an abnormal femoral head position. @*Conclusion@#The current study suggests that US findings evaluated in the first 3 months of life showed no statistically significant difference with radiographic findings evaluated at 1 year of life. The US parameters showed an excellent negative predictive value for abnormal femoral head position on radiographs.

The earliest timing of ultrasound in screening for developmental dysplasia of the hips

PURPOSE: The current study aimed to evaluate the results of ultrasound screening for developmental dysplasia of the hips (DDH) done at various weeks of life, to determine the earliest time that ultrasound screening can be performed reliably. METHODS: In this 17-year cohort study, all neonates who underwent ultrasound screening prior to the 12th week of life with subsequent follow-up radiography done at 1 year of life were included. The ultrasound images were evaluated according to the Graf classification, Harcke’s dynamic ultrasound screening method, and Terjesen’s femoral head coverage method. The radiographic images were evaluated according to the acetabular index and the femoral head position. The accuracy and correlation between the ultrasound findings from various weeks of life with the radiographic findings at 1 year of life were evaluated. RESULTS: A total of 348 neonates were included in the study, of whom 92 had abnormal ultrasound findings and 42 had abnormal radiographic findings at 1 year. Significant differences were identified between the findings of ultrasound screening examinations performed prior to the fourth week of life (day 21 and before) and the radiographic findings at 1 year of life (P0.05). The accuracy of ultrasound screening was 79.2% or higher when performed during or after the fourth week of life (day 22 and beyond). CONCLUSION: The earliest that ultrasound screening for DDH can be performed reliably is during the fourth week of life (day 22 and beyond).

Exosomes from human umbilical cord mesenchymal stem cells at passage 2 and 5:a comparative study on microRNA profiles / 中国组织工程研究

BACKGROUND: Exosomes have the function of some mesenchymal stem cells. Understanding the substance composition that plays a representative role in mesenchymal stem cell exosomes will provide clues for further exploration of synthetic exosome analogues. OBJECTIVE: To investigate the difference of microRNA expression profiles in exosomes derived from passage 2 and 5 human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS: Exosomes in the supernatant of passage 2 and 5 hUC-MSCs were extracted by ultra-high speed centrifugation. The established library was sequenced by using high-throughput sequencing technology. Then we analyzed the sequence results so as to understand the microRNA expression between different groups, and finally did a cluster analysis. RESULTS AND CONCLUSION: 427 657 kinds of microRNAs were detected in the exosomes from passage 2 hUC-MSCs, accounting for 68.93% of the total microRNAs detected; and 119283 microRNAs were detected in the exosomes from passage 5 hUC-MSCs, accounting for 19.22% of the total microRNAs detected. There were 73 526 microRNAs shared between the exosomes from passage 2 and passage 5 hUC-MSCs, accounting for 11.85% of the total microRNAs detected. Bioinformatics analysis (cluster analysis) results showed that these miRNAs were likely to be involved in 161 biological processes, including cell repair, immune and anti-aging. The microRNAs in exosomes from passage 2 to passage 5 hUC-MSCs were largely different. Partial miRNAs exhibited significantly reduced copy numbers. The top five microRNAs with a higher amount, including has-miR-146a-5p, has-miR-191-5p, has-miR-493-3p, has-miR-423-5p, and has-miR-134-5p, have the potential to be the component of synthetic exosome analogues.

A comparative study on serum-free and serum culture methods of human umbilical cord mecenchymal stem cells / 中国组织工程研究

BACKGROUND: Studies have shown increasing risks and problems in the serum culture system, such as immune rejection, batch differences and virus risk. In addition, with the discovery and application of exosomes, the serum-free culture system is becoming an increasing concern. OBJECTIVE: To compare the similarities and differences between the serum-free culture system and the traditional serum culture system, which lays the foundation for the clinical transformation of human umbilical cord mesenchymal stem cells (hUCMSCs) and provides experimental data. METHODS: Umbilical cord was collected from term infants of cesarean section under aseptic condition, and hUCMSCs were isolated and cultured by explant tissue technique. hUCMSCs was cultured with 10% fetal bovine serum (FBS) and 15% serum substitutes (AGS) from the original generation. Then an inverted microscope was used to observe cell morphological changes. Flow cytometry was used to detect cell surface markers. Cell counting kit-8 was used to detect cell proliferation. Induced differentiation experiment was used to detect cell differentiation potential. Western Blot was used to detect the protein levels of oct4, nanog and sox2. RESULTS AND CONCLUSION: Under the inverted microscope, hUCMSCs cultured with AGS showed more uniform vortex-like growth, and those cultured with FBS gradually appeared with cell differentiation or aging with the increase of cell generations. hUCMSCs cultured by both methods expressed CD73,CD90 and CD105 but lowly expressed CD34 and CD45, and there was no significant difference between the two culture methods. FBS method was superior to AGS method in proliferation ability. Results from the induced differentiation experiments showed that hUCMSCs cultured by both methods had adipogenic, osteogenic and chondrogenic abilities, and there was no significant difference between the two culture methods. hUCMSC cultured by both methods expressed oct4 and nanog but showed no significant difference in level, while the expression of sox2 was significantly higher in the hUCMSCs cultured by AGS than by FBS (P < 0.05). To conclude, the hUCMSCs cultured with AGS are in accordance with the international standards of mesenchymal stem cells. The AGS method as an alternative to the FBS method can become a preferred method for hUCMSCs culture.

Clinical outcomes of below knee amputations in diabetic foot patients

<p><b>INTRODUCTION</b>This study aims to evaluate the predictive factors affecting the clinical outcome of Below Knee Amputations (BKA) performed in diabetic foot patients admitted to National University Hospital (NUH) Multi-Disciplinary Diabetic Foot Team.</p><p><b>MATERIALS AND METHODS</b>This is a prospective cohort study of 151 patients admitted to the Department of Orthopaedic Surgery, NUH, for Diabetic Foot Problems (DFP) from January 2006 to January 2010. All had undergone BKA performed by NUH Multi-Disciplinary Diabetic Foot Team. Statistical analyses (univariate and multivariate analysis with logistic regression) were carried out using SPSS version 18.0, for factors such as demographic data, diabetic duration and control, clinical findings and investigations, indications for surgery, preoperative investigations and evaluation, microbiological cultures, and these were compared to the clinical outcome of the patient. A good clinical outcome is defined as one not requiring proximal re-amputation and whose stump healed well within 6 months. The ability to ambulate with successful use of a prosthesis after 1 year was documented. Statistical significance was set at P <0.050.</p><p><b>RESULTS</b>Mean age of study population was 55.2 years with a male to female ratio of about 3:2. Mean follow up duration was 36 months. Of BKAs, 73.5% gave a good outcome. Univariate analysis showed that smoking, previous limb surgery secondary to diabetes, high Total White Count (TW), Erythrocyte Sedimentation Rate (ESR), C-Reactive Protein (CRP), Urea, Creatinine (Cr), Neutrophils, absence of posterior tibial and popliteal pulses, low Ankle Brachial Index (ABI) and Toe Brachial Index (TBI) were associated with poor clinical outcome. Multivariate analysis showed that high CRP, ESR, Neutrophils, absence of popliteal pulse and low ABI were associated with poor clinical outcome. Of patients, 50.3% attained mobility with prosthesis after 1 year. Mortality rate was 21.2% within 6 months of operation, with sepsis being the most significant cause of death.</p><p><b>CONCLUSION</b>Success rate of BKA was 73.5%, with mortality rate being 21.2% within 6 months. In this cohort, 50.3% were able to attain eventual mobility with prosthesis after 1 year. Sepsis was the most significant cause of death. Markers of infection such as high CRP, ESR, neutrophils; and indicators of poor vascularity such as absence of popliteal pulse and low ABI were significantly associated with poor clinical outcome.</p>

Short-term effects of citrate on markers of bone metabolism in Chinese blood donor volunteers / 中国实验血液学杂志

This study was purposed to investigate the short-term effects of citrate administration on bone metabolism in the healthy blood donor volunteers. A crossover, placebo-controlled trial were conducted on 22 healthy blood donor volunteers. The volunteers received either a standardized infusion of citrate at 1.5 mg/(kg.min) or the equal volume of placebo normal saline, were washout for 2-3 weeks. During washout serial blood samples were collected and analyzed for bone biochemical markers and electrolytes, such as bone formation marker osteocalcin (OC), bone resorption marker carboxyterminal telopeptide of type I collagen (CTX), intact parathyroid hormone ((i)PTH), ionized calcium ((i)Ca(2+)) and phosphorus (P(i)). Serial urine samples were collected and analyzed for Ca(2+), P(i) and creatinine concentration. The results showed that compared with placebo group, infusion of citrate increased serum levels of OC and CTX (p < 0.0001). The greatest increase of OC and CTX levels occurred at the completion of the intervention. The increment of CTX was higher than OC (p = 0.02), and the OC/CTX ratio decreased (p < 0.01). Infusion of citrate also induced profound increase in serum (i)PTH level (p < 0.0001) and urinary calcium excretion (p < 0.0001), and decrease in serum (i)Ca(2+) (p < 0.0001) and P(i) (p < 0.01) levels. The decrease of (i)Ca(2+) level in female was higher than that in male (p = 0.007), but the changes of (i)PTH, OC, and CTX levels showed no differences between female and male. Changes of OC and CTX levels were closely related to each other (r = 0.56, p < 0.0001) and changes of both markers were negatively correlated with the change of serum (i)Ca(2+) concentration during the citrate intervention(r(OC) = -0.44, r(CTX) = -0.44, p < 0.0001). Increased levels of (i)PTH showed positively correlation with OC (r = 0.34, p = 0.02) and borderline correlation with CTX (r = 0.29, p = 0.06) in male. No such relationship was observed in female. All bone markers and electrolyte levels returned to baseline within 24 hours. It is concluded that the citrate load at the dose as a single platelet apheresis results in profound increase of bone turnover, which is characterized by a short-term increase of bone resorption and excretion of calcium. The possible effect of citrate on bone mass of long-term frequent platelet apheresis donor is worth concerning.