Objective To investigate the clinical efficacy of the treatment of long-term non-invasive positive pressure ventilation (NIPPV) combined with inhaling corticosteroids in patients with stable chronic o0bstructive pulmonary disease(COPD) complicated with respiratory failure,and to investigate the impact of longterm NIPPV combined with inhaling corticosteroids on serum levels of matrix metalloproteinase-9 (MMP-9).Methods Eighty outpatients of stable severe COPD complicated with respiratory failure divided them equally into two treatment groups (the experimental and the control groups).The two groups of patients were given oxygen therapy,inhalation of Salmeterol and fluticasone propionate powder for one year.The experimental group received additionally NIPPV therapy for 1 year.The outcomes measured included St.George's questionnaire (SGRQ) score,MMRC score,6-min working time (6-MWT),arterial partial pressure of oxygen (PaO2),partial pressure of carbon dioxide(PaCO2),Forced expiratory volume in 1 (FEV1％),and the serum levels of MMP-9 before and after treatment,and frequency of acute exacerbations of COPD and hospital says in the last one year and the following 12 months.Results After 1 year,the differences of SGRQ score,MMRC score,6-MWT,PaO2,PaCO2,FEV1％,MMP-9 in the experimental group ((63.38 ±4.46) vs.(52.93 ±4.30),t =10.67,P =0.00;(3.60±0.50) vs.(2.40 ±0.50),t =10.82,P=0.00;(159.90 ±6.50) m vs.(247.10±9.66) m,t=47.39,P=0.00;(56.85 ± 1.67) mm Hg vs.(66.10 ±2.59) mm Hg,t =10.67,P =0.00;(60.38 ±3.58)mm Hgvs.(51.88 ±3.05)mm Hg,t=10.82,P=0.00; (38.93 ±3.22)％ vs.(42.12 ±3.11)％,t=47.39,P =0.00;(182.58 ±6.60) μg/L vs.(171.73 ±6.19) μg/L,t =7.58,P =0.00) were statistically significant compared to the control group ((63.88 ± 4.88) vs.(54.30 ± 4.13),t =8.77,P =0.00; (3.65 ± 0.48) vs.(2.70±0.46),t =8.97,P =0.00;(157.98 ±5.97) m vs.(218.08±13.12) m,t =26.38,P=0.00;(56.65 ±1.51)mm Hg vs.(62.60 ± 1.91)mm Hg,t=8.77,P=0.00; (60.20 ±3.52)mm Hg vs.(56.25 ±3.09)mm Hg,t =8.97,P =0.00; (38.93 ±2.96) ％ vs.(40.70 ±3.27)％,t =26.38,P =0.00; (180.55 ±4.78) μg/L vs.(173.05 ± 5.28) μg/L,t =6.66,P =0.00).The frequency of acute exacerbations of COPD and hospital stay days were significantly decreased in the experimental group than in the control group.Conclusion Long-term NIPPV combined with inhaling corticosteroids could significantly improve the quality of life and lower the serum levels of MMP-9 of patients with severe stable COPD complicated with respiratory failure.

Objective To study the preventive and therapeutic effects of blood pressure control on hematoma expansion and neurological function in patients with ultra-early basal ganglia intracerebral hemorrhage.Methods From November 2009 to November 2011,120 patients with ultra-early basal ganglia intracerebral hemorrhage from our Hospital were enrolled and randomly divided into intensive blood pressure reduction group and general blood pressure reduction group in equal numbers (n =60).The antihypertensive agent were used intravenously to reduce the systolic blood pressure by 130-140 mm Hg within l hour after treatment in patients of intensive blood pressure reduction group; and the general blood pressure reduction group was control by 160-180 mm Hg.The blood pressure of patients in both groups was maintained for 24 hours.The volume of haematoma in CT was measured before and 24 hours after treatment.The National Institutes of Health Stroke Scale (NIHSS) score was assessed 24 hours before and after treatmentand 14 days after treatment respectively.Statistical analyses were conducted.Results Between 24 hours before and after treatment,therewere significant difference in the hematoma volume((11.99 ± 6.90) ml vs.(14.74 ± 7.75) ml,t =2.049,P =0.043) and the number of cases of hematoma enlargement(5 vs.14,x2 =5.07,P =0.024) between the two groups.Between 24 hours before and after treatment,there was no significant difference in NIHSS scale in intensive blood pressure reduction group ((9.74 ± 4.49) vs.(9.25 ± 4.10),P ＞ 0.05).Between 24 hours before and 2 weeks after treatment,there were significant difference in NIHSS scale in both groups ((9.74 ± 4.49) vs.(6.28 ± 3.68),P ＜ 0.05 ; (9.50 ± 4.81) vs.(7.82 ± 4.28),P ＜ 0.05,respectively).At two weeks after treatment,there was significant difference in NIHSS scale between two groups ((6.28 ± 3.68) vs.(7.82 ± 4.28),P ＜ 0.05).Conclusion Intensive reduction of blood pressure is safe for the treatment of ultra-early basal ganglia intracerebral hemorrhage and reduce the incidence of hematoma enlargement and improve patient's early neurological function.

Objective To evaluate the clinical value of quantitative fluorescent polymerase chain reaction (QF-PCR) in rapid prenatal diagnosis of aneuploidies. Methods Twenty-two short tandem repeats (STR) and AMXY located on chromosome 13,18,21,X and Y were used as markers to examine 1740 samples from high risk pregnant women in Down syndrome screening and advanced maternal age(≥35 yrs) by QF-PCR.Samples were also tested by karyotype analysis and the results of the two methods were compared. Results Karyotype analysis and QF-PCR results were successfully obtained from 1690 samples. All QF-PCR reports were obtained within 48 hours after sample collection.For 1639 samples,normal results were obtained by both karyotype analysis and QF-PCR.Among 51 samples that were found abnormal by karyotype analysis,41 were abnormal in QF-PCR.The rapid tests found all numerical abnormalities involving chromosome 21,18,13,X and Y in prenatal diagnosis,including trisomy 21 (n =30),trisomy 18 (n =6),45,XO (n =1 ),47,XYY (n=1),47,XXX (n=1),69,XXX (n=1) and mosaic 47,XXY[94]/46,XX[6] (n=1)(47,XXY in QF-PCR).No false positive results were found.The results obtained by QF-PCR were consistent with those of cytogenetic studies in 99.4％ of the samples (1680/1690).Only ten cases of mosain and structural abnormality could not be found (0.6％,10/1690) by QF-PCR. Conclusions Rapid QF-PCR test might diagnose all aneuploidies involving chromosome 21,18,13,X and Y.It could provide rapid and accurate diagnosis for 99.4％ pregnant women with positive Down syndrome screening and advanced maternal age.

BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.

Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.

Objective:To investigate genetic variation profiles of δ-globin (HBD gene) and hematological phenotypes in Guangdong population.Methods:Retrospective case analysis was performed in this study. Blood samples of 11 616 couples who participated in free thalassemia screening in Guangzhou from July 2020 to December 2022 were collected which underwent blood routine tests and hemoglobin (Hb) capillary electrophoresis. According to the results, 154 samples were enrolled in this study: (1)group of 35 cases with HbA 2 <2.0% but no HbF band; (2)group of 64 cases with HbA 2 < 2.0% and HbF band; (3)group of 25 cases with HbA 2 <2.0% and suspected HbA 2 variants; (4) group of 25 cases with HbA 2 ≥2.0% and <3.5% and HbF band, as well as abnormal blood routine report [mean corpuscular volume (MCV) <82 fl and/or mean corpuscular hemoglobin (MCH) <27 pg]; (5)group of 5 cases with HbA 2 ≥2.0% and <3.0% accompanied with β thalassemia gene carriers Sanger sequencing was used to detect single nucleotide variants of δ-globin. Results:(1) A total of 22 genetic variations were detected, including 6 de novo variations, and the top 3 genetic variations were respectively c.-127T>C (57.02%, 65/114), c.-80T>C (9.65%, 11/114), c.349C>T (7.89%, 9/114). (2) In group of patients with HbA 2 <2.0% but no HbF band, 22 cases (62.85%, 22/35) had HBD gene variation, including 7 cases with MCV and MCH lower than reference values, 4 cases with α thalassemia; 13 cases had no HBD gene variation, including 12 cases with lower MCV and MCH. Among 19 cases with abnormal blood routine test results, levels of HbA 2 in patients (7 cases) with HBD gene variation were lower compared with those without HBD gene variation (12 cases) ( P<0.01%). (3)In group of patients with HbA 2<2.0% with HbF band, 59 cases (92.18%, 59/64) had HBD gene variations whose mutations all occurred in promoter region, and the HbF were all lower than 5.0%; 5 cases with HbF >5.0% had no HBD gene variation. (4) In group of patients with HbA 2 <2.0% and suspected HbA 2 variants, the detection rate was 100% (25/25) and δ-globin variants <1.0%. (5) In group of patients with HbA 2 ≥2.0% and <3.5% and HbF band accompanied with abnormal blood routine results, no HBD gene variation was found. (6) In group of 5 patients with HbA 2 ≥2.0% and <3.0% with β thalassemia gene carriers, HBD gene variation were found in all cases, and the level of HbA 2 was (2.62±0.17)% and HbF was (3.62±2.22)%. Conclusions:There are various genotypes of HBD gene variation, among which HBD: c.-127T>C is the most common in Guangdong population in China. Mutations in the promoter region may cause decrease in HbA 2 and increase in HbF which is mostly less than 5% but exceeds 5.0% when combined with β thalassemia. Our study enriched the gene mutation profiles of HBD gene in Guangdong population.

OBJECTIVETo determine the immune repertoires of peripheral CD4+T cell receptor (TCR) Vb CDR3 in primary biliary cirrhosis (PBC) and analyze TCR diversity and preferred usage at sequence-level resolution.

METHODSARM-PCR and high-throughput sequencing were used to obtain millions of TCR Vb CDR3 sequences from peripheral CD4+T cells isolated from 7 patients with PBC and healthy volunteers. All sequencing data were analyzed, together with corresponding clinical information, by bioinformatic software. The Mann-Whitney U test was used for statistical analysis.

RESULTSThe PBC patients showed a lower level of diversity among the peripheral CD4+TCR Vb CDR3 than the healthy volunteers, and patients with higher level progression of the disease showed a greater lack of diversity. In addition, 4 specific preferred-usage amino acid sequences were discovered for the PBC patients: ASSFTGGPVEQY, ASSLISSGNNEQF, ATSRDTLAGGPGDTQY, and SASLEGNTEAF; these sequences were also found in higher frequencies in patients with later stages of PBC.

CONCLUSIONSDecreased TCR Vb CDR3 diversities and specific preferred usage of TCR CDR3 sequences in peripheral CD4+T lymphocytes in PBC suggest that clonal expansion of a large number of CD4+T cells may be an important factor for PBC progression. These data provide a better understanding about the general characteristics of CD4+T cells in PBC patients and related to pathogenesis of the disease, and may provide useful insights into potential targets for immunotherapy.

Objective:To analyze the consistency between karyotyping and quantitative fluorescent- polymerase chain reaction (QF-PCR) in prenatal diagnosis.Methods:This study retrospectively analyzed the clinical data of 10 967 patients undergoing karyotyping and QF-PCR for prenatal diagnosis in Guangzhou Women and Children's Hospital from January 2010 to December 2017. The failure rate, results, and diagnosis of common chromosomal disorders of the two methods were compared. The sensitivity and specificity of QF-PCR in detecting chromosomal mosaicism were evaluated using the receiver operative characteristic (ROC) curve.Results:(1) The failure rates of karyotyping and QF-PCR were 0.99% (109/10 967) and 0.10% (11/10 967), respectively. (2) The karyotypes of 9 960 out of the 10 858 successfully cultured samples were normal, and 99.89% (9 949/9 960) results were consistent between the two methods. The other 898 cases included 694 (77.28%) with common chromosomal abnormalities (trisomy 21, 18 and 13 and sex chromosomal abnormality) and 204 (22.72%) with other chromosomal abnormalities. The consistency between the two methods in detecting common chromosomal abnormalities was 95.68% (664/694). (3) The consistency in the detection of trisomy 21, 18 and 13 and sex chromosomal abnormality between karyotyping and QF-PCR were 99.74% (382/383), 100.00% (125/125), 100.00% (33/33) and 81.05% (124/153). However, the common chromosomal mosaicism was only noted for 44.44% (24/54). (4) Among cases with a mosaic ratio over 18.5%, the sensitivity and specificity of QF-PCR were 0.958 (95% CI: 0.789-0.999) and 0.600 (95% CI: 0.406-0.773) with the area under the ROC curve (AUC) of 0.811 (95% CI: 0.696-0.926, P<0.001). (5) Thirty cases with negative QF-PCR results but positive mosaic chromosomal aberrations were followed up. Ten (33.3%) pregnant women terminated their pregnancies, and two (6.7%) were lost to follow-up. The other 18 cases delivered healthy neonates that all survived after birth. Conclusions:In prenatal diagnosis, QF-PCR and karyotyping were highly consistent in the detection of trisomy 21, 13, and 18, but have significant discordance in the diagnosis of sex chromosomal abnormality.

With the development of precision medicine and individualized treatment, tissue biopsy in cancer patients diagnosis and therapy has been broadly used. However, because it’s hard to collect enough samples for biliary tract tumors, liquid biopsy was broadly applied for the diagnosis. In liquid biopsy, circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes carrying tumor-specific information are released from tumor tissue into blood, bile, and other body fluids, which makes tumor biopsy samples easily to be obtained in a non-invasive way. At the same time, through a series of morphological and molecular measurements as well as genetic characterization, liquid biopsy can be used to look for the new early diagnostic markers, and therapeutic targets, monitoring progression and prognosis of diseases. This article outlined the current technology used to detect circulating tumor cells, circulating tumor DNA, and tumor-derived exosomes, and summarizes the latest advances in the clinical application of liquid biopsy in biliary tract cancers.