ObjectiveTo establish a set of single nucleotide polymorphisms (SNP) detection protocol for inbred rats based on multiplex PCR-ligase detection reaction (LDR). MethodsA total of 40 rats SNP sites were selected on chromosomes 1-20 and X of rats among 5 inbred strains of rats, and the 40 SNP sites were randomly divided into four groups. A genetic detection protocol for 4 groups of SNP in inbred rats based on multiplex PCR-LDR technology was constructed. 9 commonly used rat strains from two other domestic rat suppliers were detected by this protocol. Finally, the feasibility of this protocol was verified by comparing the amplification effects of different DNA polymerases by a third-party laboratory. ResultsWhen using the constructed SNP detection protocol for inbred rats to test 5 rat strains, all sites in each sample obtained good amplification results. The 9 commonly used rat strains from two other rat suppliers in china were also well amplified by this SNP detection protocol, and 40 SNPs were homozygous in each Inbred strain. The results of detection of the same rat DNA samples with three different DNA polymerases showed that the Multiplex PCR Kit, AmpliTaq Gold 360 DNA polymerase and Platinum II Taq hot start DNA polymerase had electrophoretic peaks of amplification products at all SNP sites in groups 1 to 3, and Platinum II Taq hot start DNA polymerase had one less electrophoretic peak of the amplification products at the SNP sites in group 4. In addition, inter-laboratory comparisons showed consistent results for the same amplification system. ConclusionBased on multiplex PCR-LDR technology, this study successfully established a SNP detection protocol for rats covering all autosomes and X chromosomes with the excellent stability and repeatability.

OBJECTIVES: The prevalence rate of sleep problems in children in China is increasing in recent years. There are inconsistencies in the relationship between physical activity and sleep, and the impact of screen time on young children seems more obvious.This study aims to understand the current situation of outdoor activity time, screen time and sleep (total sleep duration, bedtime, and wake-up time) and the associations between outdoor activity time and screen time with sleep for children aged 36-54 months in Kaifu District of Changsha. METHODS: Using the cluster sampling method, 1 286 newborns delivered in 3 community health service centers in Kaifu District, Changsha from January to December 2015 were selected as the research subjects to establish a prospective birth cohort. According self-designed questionnaire and household follow-up to select 36, 42, 48, 54 months Children's data. The mixed linear model was used to explore the associations of outdoor activity time and screen time with sleep. RESULTS: The 36-54 months children's total sleep duration was decreased from 11.60 h/d to 10.92 h/d (P<0.001); bedtime time delayed from 21:58 to 22:00 (P=0.124); wake-up time advanced from 7:52 to 7:37 (P<0.001); outdoor activity time was decreased from 2.58 h/d to 1.94 h/d (P<0.001), and screen time was decreased from 1.28 h/d to 1.09 h/d (P<0.001). With aging, the prevalence of sleep <10 h/d and outdoor activity time <2 h/d was increased significantly, and the prevalence of screen time ≥1 h/d was decreased (P<0.05). Mixed linear models showed that longer outdoor activity time was related to increased total sleep duration (β=0.22, 95% CI 0.00 to 0.44) and delayed wake-up time (β=0.16, 95% CI 0.00 to 0.32), and longer screen time was related to delayed bedtime (β=0.22, 95% CI 0.05 to 0.39) and wake-up time (β=0.24, 95% CI 0.08 to 0.41). These associations were different in gender. Longer outdoor activity time was related to delayed wake-up time (β=0.37, 95% CI 0.14 to 0.59) in boys, but not in girls (β=-0.16, 95% CI -0.33 to 0.01). Longer screen time was associated with delayed bedtime (β=0.40, 95% CI 0.09 to 0.63) and wake-up time (β=0.33, 95% CI 0.15 to 0.51) in girls, but only related to delayed wake-up time (β=0.29, 95% CI 0.06 to 0.52) in boys, and the degree of association was lower than that of girls. CONCLUSIONS Among children aged 36-54 months in Kaifu District, Changsha, there are problems including going to bed late and getting up late, insufficient physical activity, and long screen time. Outdoor activity time and screen time are related to sleep. Increasing outdoor activity time and reducing screen time can help to improve children's sleep.
Infant, Newborn ; Male ; Child ; Female ; Humans ; Child, Preschool ; Screen Time ; Cohort Studies ; Birth Cohort ; Prospective Studies ; Sleep

Objective:To explore application status and development trend of spinal muscular atrophy (SMA) genetic diagnosis technology based on the national rare diseases registry system of China.Method:A total of 200 SMA children registered at the Capital Institute of Pediatrics from July 2016 to December 2018 were included in this retrospective cross-sectional survey. The basic data, clinical subtypes, genotypes, and related genetic testing information of SMA children were obtained by checking SMA registration information, genetic testing reports, and also by telephone follow-up. The patient number and the composition of different genetic diagnosis technologies were analyzed by the stratification of genetic testing at various time. The correlation between the proportion of genetic diagnosis technology and genetic testing time was analyzed with Pearson correlation analysis.Result:There were 3 SMA cases with incomplete data, the remaining 197 SMA cases were included in this study. There were 37 (18.8%), 115 (58.4%) and 45 (22.8%) patients with type Ⅰ, Ⅱ and Ⅲ SMA, respectively. There were 185 cases of SMN1 homozygous deletion (93.9%), and 12 cases with compound heterozygotes (6.1%). Seven SMA-related genetic technologies were used from 2004 to 2017. MLPA accounted for 54.1% (100/185) used approach, followed by PCR-RFLP and first-generation sequencing, which accounted for 22.7% (42/185) and 10.3% (19/185), respectively. Nine, 6, 5 and 4 cases were tested with AS-PCR, qPCR, WES and DHPLC, respectively (2.2%-4.9%). The proportion of MLPA increased gradually since 2010 ( r=0.95, P<0.05), while PCR-RFLP declined gradually since 2004 ( r=-0.99, P<0.05). No correlation was found between technology and testing time for other genetic testing technologies ( P>0.05). The proportion of quantitative genetic technologies (MLPA, qPCR and DHPLC) increased gradually since 2010 ( r=0.94, P<0.05), and qualitative genetic technologies (PCR-RFLP, first-generation sequencing, AS-PCR and WES) decreased gradually since 2004 ( r=-0.94, P<0.05). The duplication detection rates of homozygous deletion and compound heterozygous mutation were 12.4% (23/185) and 41.7% (5/12), respectively (χ 2=5.86, P<0.05). During 2008-2015, the proportion of "the reports of both copy numbers of SMN1 gene and SMN2 gene" increased from 56.8% (21/37) in 2008-2015 to 69.1% (56/81) in 2016-2017. Conclusion:Genetic diagnosis of SMA has gradually developed from qualitative detection technology to quantitative detection technology, such as MLPA and qPCR, in China. In more and more SMA quantitative test reports, quantitative results of SMN2 gene are also provided in addition to quantitative results of SMN1 gene.

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.
COVID-19 ; Humans ; Medicine, Chinese Traditional ; Research ; SARS-CoV-2 ; Treatment Outcome

Objective To observe aerobic exercise on blood lipid and intercellular adhesion molecule-1 (ICAM-1),lymphocyte function-associated antigen-1 (LFA-1) in hyperlipidemia rats.Methods 120 male Sprague Dawley (SD) rats were randomized into the 4 groups (n =30):normal diet group (control group),high fat group (HF group),HF + SBC-115076 group [SBC-115076,8 mg/kg · w)] and HF + aerobic exercise group.Rats in HF group and HF + SBC-115076 group did not exercise during feeding and lived in normal cage.Rats in HF + SBC-115076 group recieved PCSK9 inhibitor SBC-115076 (8 mg/kg) injection once a week for 8 weeks.HF + aerobic exercise group underwent load-free swimming training 6 days a week for 8 weeks.The rats were sacrificed 8 weeks later.The serum levels of triglyceride (TG),total cholesterol (TC),low density lipoprotein (LDL),high density lipoprotein (HDL),ICAM-1 and LFA-1 were measured by apical blood sampling.Hematoxylin-eosin (HE) staining of thoracic aorta to observe pathological changes of aorta.Results After modeling,there was no significant difference in the food intake levels of the hyperlipidemia rats (P ＞ 0.05).The body weight of HF rats and HF + SBC-115076 rats increased significantly than HF + aerobic exercise rats (P ＜ 0.01).The levels of serum lipid profile,ICAM-1,and LFA-1 were significantly different between HF rats and control rats.Compared with HF rats,serum levels of TG,TC,LDL,ICAM-1 and LFA-1 in HF + SBC-115076 group and HF + aerobic exercise group were significantly lower and HDL levels were significantly higher.HE staining showed that the intimal thickening was significantly improved in HF + aerobic exercise group with less endothelial damage.Conclusions Aerobic exercise can reduce the levels of serum TG,TC,LDL and increase HDL level in rats with hyperlipidemia.It can improve the intimal thickening and endothelial damage caused by high-fat diet by reducing the expression levels of serum ICAM-1 and LFA-1,and has anti-atherosclerosis effect.

[Purpose]To summarize Professor SHI Weiqun's clinnical experience in cirrhosis by Huayu method of traditional Chinese medicine, who is the famous doctor of traditional Chinese medicine in Zhejiang Province.[Method] To study and research Professor SHI Weiqun' s subjects and books on cirrhosis, to learn revelant theoretical knowledge, to follow his guidance, to collect and arrange typical cases record, and his methods of TCM differentiation of symptoms and signs^by these methods to explore the thinking and syndrome differentiation of the syndrome, summrize Professor SHI Weiqun's clinnical experience in cirrhosis by Huayu method of traditional Chinese medicine.[Result]Professor SHI Weiqun thinks main points of pathogenesis is blood stasis, but, when treating individuals, need to combine the difference among Qi, blood, Yin and Yang, and form the academic experience of Xingqi Huoxue, Yiqi Huayu, Yangxue Huayu, Wenyang Huayu, Ziyin Huayu.He often selects Chaihu Shugan powder, Buzhong Yiqi decoction, Chinkuei Shin Chewan pills, Liuwei Dihuang Wan, Erzhi pills, Xuefu Zhuyu decoction ect.[Conclusion]Professor SHI Weiqun has rih clinical experiences, merges ancient and modem, makes good use of activating blood circulation drugs, selects precise and appropriate, has formed a relatively rigorous thought of diagnosis and treatment, the academic thought and clinical experience is worthy of inheritance and promotion.

Hepatitis B virus (HBV) infection is a worldwide health problem and vertical transmission is the main source of chronic infection in Asian countries. Breakthrough perinatal HBV transmission occurs in 10%-15% of infants born to pregnatal women with high viral load even with appropriate postexposure immunoprophylaxis. Management of chronic hepati?tis B (CHB) during pregnancy remains a challenge, with unique issues that involve prevention of mother-to-child transmis?sion (MTCT) and safe use of antiviral therapy. Previous studies indicate that antiviral therapy with analogs (Lamivudine, Tel?bivudine or Tenofovir) may prevent vertical transmission during late pregnancy in highly viremic mothers. This article sum?marized relevant studies that antiviral therapy in preventing MTCT of hepatitis B virus.

Objective To explore the curative effect of traditional Chinese herbal packet combined with ST 36 injection therapy on gastrointestinal dysfunction in severe patients. Methods Sixty severe patients with gastrointestinal dysfunction were randomly divided into the control group and treatment group with random digital number, 30 cases in each group. The control group received conventional care and besides conventional care , the treatment group was treated with traditional Chinese herbal packet combined with ST 36 injection. The two groups were compared 7 days after treatment in terms of scores by (acute physiology and chronic health evaluationⅡ, APACHEⅡ) and improvement of gastrointestinal dysfunction. Result After treatment, the APACHE II score and improvement of gastrointestinal dysfunction of the treatment group were significantly higher and better than those of the control group (P<0.05). Conclusion The traditional Chinese herbal packet combined with ST 36 injection can promote the recovery of gastrointestinal dysfunctional in severe patients.

OBJECTIVETo investigate the different outcomes by dexamethasone in adults immune thrombocytopenia purpura (ITP) with different types of platelet specific-autoantibodies.

METHODSA total of 185 ITP were enrolled, 61 males and 124 females, with a median age of 42 (18-83) years, including 117 newly diagnosed, 35 persistent, and 33 chronic cases. All the patients received the dexamethasone at an initial dose of 40 mg per day for 4 days and a low dose of 5-10 mg for 3-4 weeks. The platelet specific-autoantibodies were identified by the modified monoclonal antibody-specific immobilization of platelet antigen (MAIPA) assay.

RESULTSAmong the IgG positive patients, the response rates in anti-GPIIb/IIIa antibody, anti-GPIbα antibody, both antibody positive, and both antibody negative were 87.5%, 50.0%, 68.0%, and 72.3% (χ²=11.489, P<0.05), respectively. Among the IgM positive patients, the response rates in the four groups were 82.1%, 71.4%, 61.9%, and 68.9% (χ²=2.719, P=0.437), respectively. Among the GPIbα antibody positive patients, the response rates in IgG alone, IgM alone, both positive, and both negative were 52.4%, 59.1%, 76.1%, and 77.9% (χ²=10.811, P<0.05), respectively. Among the GPIIb/IIIa antibody positive patients, the response rates in the four groups were 73.3%, 71.0%, 78.6%, and 66.3% (χ²=1.374, P=0.719), respectively.

CONCLUSIONITP patients with GPIbα-IgG antibody have worse response to dexamethasone treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; Autoantibodies ; Blood Platelets ; Dexamethasone ; Female ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; Purpura, Thrombocytopenic, Idiopathic ; Young Adult

Adrenal Gland Diseases ; diagnosis ; Female ; Humans ; Middle Aged ; Retroperitoneal Space