ObjectiveTo investigate the genetic polymorphism and structure of 47 autosomal microhaplotypes in the Han population in Changshu City, Jiangsu Province, and to evaluate the forensic efficiencies and forensic parameters. MethodsThe DNA library of unrelated individual samples was prepared according to MHSeqTyper47 kit manual and sequenced on the MiSeq FGx platform. Microhaplotype genotyping and sequencing depth statistics were processed using MHTyper. The genetic information of samples was then evaluated. The fixation index and genetic distance between the Jiangsu Changshu population and the reference populations in the 1000 Genomes Project phase 3 (1KG) were calculated, and forensic parameters were evaluated. ResultsThe fixation index and genetic distance between the Han population in Changshu, Jiangsu, and the CHB (Han Chinese in Beijing, China) reference population in 1KG were the lowest. The effective allele number (A_e) of each locus is also the closest between the two populations. The combined matching probability (CMP) of the Changshu Han population is close to the 5 populations of the East Asian reference super-population in 1KG, which is 1.25×10^-36, and the combined probability of exclusion reached 0.999 999 999 964 1. ConclusionThis study reported the genetic polymorphism and allele frequency of 47 microhaplotypes in a Han population in Changshu City, Jiangsu Province. This information provides a data basis for 47 microhaplotypes in forensic applications. In addition, the polymorphism differences between the 1KG reference population and the Han population in Changshu, Jiangsu were compared, and the genetic structure of 47 microhaplotypes in the Han population in Changshu, Jiangsu was revealed. In general, the reference data of the East Asian super-population in 1KG is more in line with the genetic characteristics of Han population in Changshu, Jiangsu.

Objectives To explore the feasibility of temporary veno-venous extracorporeal membrane oxygenation(VV-ECMO)technology for early on-site treatment,through establishing an animal model of severe blast lung injury in goats by free-field chemical explosion experiments in high-altitude regions.Methods A total of 16 adult goats were selected,and divided into the control group and the treatment group according to the random number table method,with 8 goats in each group.A model of severe blast lung injury was established at an altitude of 4 600 meters above sea level,then the goats in the control group were given respiratory support and the goats in the treatment group were given temporary VV-ECMO treatment.The survival status of the goats 15 minutes after injury was recorded,the vital signs[including body temperature,respiration rate,heart rate,and mean arterial pressure(MAP)]and arterial blood gas analysis indicators[including pH,arterial partial pressure of oxygen(PO2),arterial partial pressure of carbon dioxide(PCO2),oxygen saturation(SaO2),lactate(LAC),calcium(Ca2+),hematocrit(HCT),and hemoglobin(Hb)]before injury and 1 hour,2 hours,3 hours after injury were compared in the two groups.The post-mortem examination was performed on all dead goats and sacrificed goats after treatment,the severity of lung injury was assessed by organ injury scaling(OIS),and the lung injury score was evaluated by abbreviated injury scale(AIS).The wet-to-dry weight ratio(W/D)and lung coefficient were calculated.Results Within 15 minutes after the explosion,4 goats in the control group died and 4 goats survived;and 5 goats in the treatment group died and 3 goats survived.There was no statistically significant difference in the body temperature,respiration rate,heart rate,or MAP before and after injury between the two groups(P>0.05).The PaO2 and SaO2 1 hour,2 hours,and 3 hours after injury in the treatment group were superior than those in the control group(P<0.05),the Ca2+ 2 hours after injury was significantly higher than that in the control group(P<0.05),and there was no statistically significant difference in the pH,PCO2,LAC,HCT or Hb at different time points after injury between the two groups(P>0.05).There was no statistically significant difference in the OIS,AIS or lung coefficient between the two groups(P>0.05),but the W/D of the lung tissue in the control group was lower than that in the treatment group(P<0.05).Conclusion We have established a novel,feasible,and stable treatment effect temporary VV-ECMO animal treatment strategy for the first time in the high-altitude regions,which can provide animal experiment evidence for the early on-site VV-ECMO treatment of severe blast lung injury in high-altitude regions.

Aim To investigate the mechanism of treatment of hepatolenticular degeneration with Garde-nia and Rhubarb decoction based on network pharma-cology and animal experiments.Methods The chemi-cal components and target sites of Gardenia and Rhu-barb decoction were retrieved using the Traditional Chi-nese Medicine System Pharmacology Analysis Platform(TCMSP).The disease targets were obtained by searching the databases of DisGeNET,GeneCards.The PPI network of active compound target sites was constructed using the STRING database.GO function and KEGG pathway enrichment analysis were per-formed using the DAVID database to predict the action pathway.A copper-loaded WD rat model was estab-lished by intragastric administration of copper sulfate pentahydrate.A total of 36 rats were randomly divided into the normal group,model group,penicillamine group and the low,medium and high dose groups of gardenia rhubarb.The relevant indicators and patho-logical changes of liver tissue were detected in WD rats.Results Network pharmacology screening identi-fied 68 potential active components of Gardenia and Rhubarb Decoction,30 intersection targets of diseases and drugs,involving key targets such as TNF,IL10,IGF1,IL1B,TP53,CASP3,PPARG,IL6,CXCL8,IL1A,TGFB1,mainly related to signal pathways such as MAPK,AGE-RAGE signaling pathway in diabetic complications.In the animal experiments,compared with the normal group,the urine copper,liver copper,blood copper,liver coefficient,serum and liver ALT,AST,TNF-α,CASP3,P53 levels in the model group rats significantly increased(P＜0.05);hepatocyte swelling,cytoplasm loose reticular appearance,feath-er-like degeneration and reticular necrosis were ob-served in liver tissue pathology;compared with the model group,the urine copper,liver copper,blood copper,liver coefficient,serum and liver ALT,AST,TNF-α,CASP3,P53 levels in the penicillamine group and Gardenia and Rhubarb decoction groups were sig-nificantly reduced(P＜0.05),and the degree of re-ticular necrosis in the rat liver cells in the penicilla-mine group and the Gardenia and Rhubarb decoction group was significantly reduced.Conclusions Garde-nia and Rhubarb decoction has the effect of regulating copper metabolism and reducing liver injury.The mechanism of action may be related to reversing apop-tosis and downregulating protein expression of TNF-α,CASP3,P53 in MAPK signaling pathway.

OBJECTIVE: This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province. METHODS: We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed. RESULTS: The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α ^3.7/αα (50.23%) and β ^IVS-II-654/β ^N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively. CONCLUSION Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobinopathies/genetics* ; China/epidemiology* ; High-Throughput Nucleotide Sequencing

Objective:To explore the effects and mechanism of electroacupuncture (EA) on expression of myostatin (MSTN), muscle-specific ring finger protein 1 (MuRF1/Trim63), F-box only protein 32 (Atrogin-1/ Fbxo32), myogenic differentiation antigen (Myod) and myogenin (Myog) in traumatic spinal cord injury (TSCI) rats. Methods:A total of 45 adult female Sprague-Dawley rats were randomly divided into sham operation group (n = 12) and operation group (n = 33). The TSCI model was established with the modified Allen's method. After modeling, there were 24 survival rats and they were randomly divided into model group (n = 12) and EA group (n = 12). EA group was electroacupunctured at Dazhui (DU 14), Mingmen (DU 4) and bilateral Zusanli (ST 36) for 10 minutes, once a day, six times a week for 28 days. Basso-Beattie-Bresnahan (BBB) score was tested before modeling, and three days, seven days, 14 days, 21 days and 28 days after modeling. The rats were measured their body mass before and 28 days after modeling. The ratio of gastrocnemius wet mass was calculated; the cross-sectional area (CSA) and fiber diameter were measured by HE staining; the expression of MSTN, Trim63, Fbxo32, Myod and Myog mRNA were tested with real-time quantitative polymerase chain reaction (qPCR). Results:Three days, seven days, 14 days, 21 days, and 28 days after modeling, the score of BBB was lower in the model group than in the sham operation group (P < 0.01); seven days, 14 days, 21 days, and 28 days after modeling, the score of BBB was higher in EA group than in the model group (P < 0.01). Compared with the sham operation group, the mass of rats, the gastrocnemius wet mass, the CSA and the diameter of the muscle fiber were smaller in the model group (P < 0.05), while the expression of MSTN, Trim63, Fbxo32, Myod and Myog mRNA were higher (P < 0.05). Compared with the model group, the mass of rats, the gastrocnemius wet mass, the CSA, the expression of Myod and Myog mRNA were higher (P < 0.05) in EA group, while the expression of MSTN, Trim63 and Fbxo32 mRNA were lower (P < 0.05). Conclusion:EA might delay the gastrocnemius atrophy in TSCI rats by down-regulating the expression of MSTN, Trim63, Fbxo32 mRNA and up-regulating the expression of Myod and Myog mRNA via controlling the differentiation of the muscle satellite cells and the degradation of protein in skeletal muscle cells.

BACKGROUNDMitofusin-2 (MFN2), a well-known mitochondrial fusion protein, has been shown to participate in innate immunity, but its role in mediating adaptive immunity remains poorly characterized. In this study, we explored the potential role of MFN2 in mediating the immune function of T lymphocytes.

METHODSWe manipulated MFN2 gene expression in Jurkat cells via lentiviral transduction of MFN2 small interfering RNA (siRNA) or full-length MFN2. After transduction, the immune response and its underlying mechanism were determined in Jurkat cells. One-way analysis of variance and Student's t-test were performed to determine the statistical significance between the groups.

RESULTSOverexpression of MFN2 enhanced the immune response of T lymphocytes by upregulating Ca2+ (359.280 ± 10.130 vs. 266.940 ± 10.170, P = 0.000), calcineurin (0.513 ± 0.014 vs. 0.403 ± 0.020 nmol/L, P = 0.024), and nuclear factor of activated T cells (NFATs) activation (1.040 ± 0.086 vs. 0.700 ± 0.115, P = 0.005), whereas depletion of MFN2 impaired the immune function of T lymphocytes by downregulating Ca2+ (141.140 ± 14.670 vs. 267.060 ± 9.230, P = 0.000), calcineurin (0.054 ± 0.030 nmol/L vs. 0.404 ± 0.063 nmol/L, P = 0.000), and NFAT activation (0.500 ± 0.025 vs. 0.720 ± 0.061, P = 0.012). Furthermore, upregulated calcineurin partially reversed the negative effects of MFN2 siRNA on T cell-mediated immunity evidenced by elevations in T cell proliferation (1.120 ± 0.048 vs. 0.580 ± 0.078, P = 0.040), interleukin-2 (IL-2) production (473.300 ± 24.100 vs. 175.330 ± 12.900 pg/ml, P = 0.000), and the interferon-γ/IL-4 ratio (3.080 ± 0.156 vs. 0.953 ± 0.093, P = 0.000). Meanwhile, calcineurin activity inhibitor depleted the positive effects of overexpressed MFN2 on T cells function.

CONCLUSIONSOur findings suggest that MFN2 may regulate T cell immune functions primarily through the Ca2+-calcineurin-NFAT pathway. MFN2 may represent a potential therapeutic target for T cell immune dysfunction-related diseases.

Objective To investigate the genotype of D class of carbapenemase of carbapenem－resistant Acinetobacter baumannii in traumatology department, so as to guide the rational use of antibiotics as well as prevent the nosocomial infection. Methods A total of 96 strains of Acinetobacter baumannii were collected in traumatology department. Patient history were analyzed and genotype of carbapenemases of OXA－51,OXA－23,ISAba1－oxa－51 and ISAba1－oxa－23 were analyzed by PCR.Results 70.84% (68/96) of Acinetobacter baumannii were isolated from wound tissue. 12 antibacterial agents were tested, 96 strains of Acinetobacter baumannii showed the highest resistance rate (78.13%) to imipenem and the lowest resistance rate (43.75%) to levofloxacin. Among 23 strains of imipenem－sensitive Acinetobacter baumannii, OXA－ 51 gene expression was not detected in 4 strains, however, OXA－23 gene expression were observed in 5 strains. The gene expression rates of OXA－51,OXA－23, ISAba1－oxa－51 and ISAba1－oxa－23 genes were 100%, 95.89%, 79.45% and 71.23% respectively in 73 strains of imipenem－insensitive Acinetobacter baumannii. The rate of gene expression profile of OXA－51+ISAba1－oxa－51+OXA－23+ISAba1－oxa－23 was 65.75% .Conclusion The strains of imipenem－sensitive Acinetobacter baumannii may have the gene expression of OXA－23 of D class of carbapenemase. The main pattern of carbapenemase－producing was OXA－51+ISAba1－oxa－51+OXA－23+ISAba1－oxa－23 in traumatology department,which might be regarded as one of the mechanisms that led to the high resistance of the bacterium of acinetobacter baumannii to carbapenem antibiotics.

OBJECTIVETo analyze the number of myeloid-derived suppressor cells(MDSC) and the level of prostaglandin E2(PGE2) in the bone marrow of adult ITP patients, and to explore their possible mechanisms involved in the pathogenesis of this disease.

METHODSTwenty-five patients of newly diagnosed ITP, 25 patients of complete remission group and 15 patients of control group were selected. The number of MDSC in the bone marrow between 3 groups was detect by flow cytometry (FCM). The serum level of prostaglandin E2 (PGE2) in 3 groups was determined by enzyme linked immunosorbent assay (ELISA). The relative expression of IFN-γ mRNA in bone marrow mononuclear cells was measured by real time fluorescence quantitative polymerase chain reaction (RT-qPCR) in each groups.

RESULTSThe number of MDSC in the complete remission group was significantly higher than that in the control group (P<0.05); the number of MDSC in the newly diagnosed group was higher than that in the control group; the number of MDSC in the complete remission group was higher than that in the newly diagnosed group. The serum level of PGE2 in bone marrow of ITP patients in the newly diagnosed group was higher than that of the control group（P<0.05）. The serum level of PGE2 in the bone marrow of ITP patients of the complete remission group was higher than that of the control group (P<0.05）. The level of PGE2 in bone marrow serum of ITP patients of the newly diagnosed group was lower than that in the complete remission group(P<0.05）. The relative expression level of IFN-gamma in bone marrow mononuclear cells of the ITP patients in newly diagnosed group was higher than that in the control group and the complete remission group（P<0.001）. The relative quantification (RQ) of IFN-γ in bone marrow mononuclear cells was 2.60 between the newly diagnosed group and the complete remission group.

CONCLUSIONWhen adult ITP disease is remitted, the number of MDSC rises and correlates with the therapeutic response and PGE2 level in the bone marrow.

Objective To analyze the change of thyroid function,thyroglobulin antibody and thyroglobulin after cervical lymph node dissection of thyroid carcinoma.Methods A total of 240 patients with thyroid disease were selected and treated with cervical lymph node dissection of thyroid carcinoma.The surgical effect was analyzed,and the thyroid function,thyroglobulin antibody and thyroglobulin after operation were analyzed.Results The efficacy of the observation group was significantly better than the control group (P ＜ 0.05).Three days after treatment,the levels of T3,T4,FT3,FT4 and TSH significantly decreased (P ＜ 0.05),and the decreasing degree in the control group was significantly greater than the observation group (P ＜ 0.05).Six months after operation,the levels of TgAb and Tg decreased significantly in both groups,and the level of TgAb and Tg in the observation group decreased more significantly than the control group (P ＜ 0.05).Conclusion Compared with the traditional surgical treatment for thyroid cancer,cervical lymph node dissection is more worthy of clinical application.

Objective To analyze the change of thyroid function,thyroglobulin antibody and thyroglobulin after cervical lymph node dissection of thyroid carcinoma.Methods A total of 240 patients with thyroid disease were selected and treated with cervical lymph node dissection of thyroid carcinoma.The surgical effect was analyzed,and the thyroid function,thyroglobulin antibody and thyroglobulin after operation were analyzed.Results The efficacy of the observation group was significantly better than the control group (P ＜ 0.05).Three days after treatment,the levels of T3,T4,FT3,FT4 and TSH significantly decreased (P ＜ 0.05),and the decreasing degree in the control group was significantly greater than the observation group (P ＜ 0.05).Six months after operation,the levels of TgAb and Tg decreased significantly in both groups,and the level of TgAb and Tg in the observation group decreased more significantly than the control group (P ＜ 0.05).Conclusion Compared with the traditional surgical treatment for thyroid cancer,cervical lymph node dissection is more worthy of clinical application.