Objective To analyze the changes of serum total bilirubin (TBil) and gamma glutamyltransferase (GGT) in patients with type 2 diabetes newly diagnosed without complications and explore their relationship with vascular complications of type 2 diabetes.Methods The levels of serum TBil and GGT were determined by automatic biochemical analyzer in 42 healthy person (control group),64patients with type 2 diabetes without complications (diabetes without complications group) and 81 patients with type 2 diabetes with vascular complications (diabetes with complications group).Results The levels of serum TBil and GGT were (7.89 ± 4.81) μ mol/L and (29.81 ± 14.97) U/L in diabetes with complications group.The level of serum TBil was (10.42 ± 3.55) μ mol/L in control group.The levels of serum TBil and GGT were (11.30 ± 4.16) μ mol/L and (47.00 ± 38.57) U/L in diabetes without complications group.The level of serum TBil in diabetes with complications group was lower than that in control group and diabetes without complications group (P ＜ 0.05).The level of serum GGT in diabetes with complications group was lower than that in diabetes without complications group (P ＜ 0.05).Pearson correlation analysis showed that 24-hour urinary protein excretion,urinary albumin creatinine ratio had negative correlation with serum TBil (P ＜0.05).Logistic regression analysis showed that GGT was the protective factor of diabetic vascular complications (OR =0.965).Conclusion The levels of serum TBil and GGT may be associated with the occurrence and development of vascular lesions in patients with type 2 diabetes.

Objective To investigate the effect and its significance of low-dose insulin on Nmethyl-D-aspartate receptor (NMDAR) level and its downstream signaling of c-Jun N-terminal kinase (JNK) and the extracellular signal-regulated protein kinase (ERK,p44/42MAPK) in streptozotocininduced diabetic rats.Methods Ten-week-old male SD rats were randomly divided into 3 groups:control group,diabetic group and insulin-treated diabetes group.Diabetes was induced by streptozocin (STZ,60mg/kg) injected intraperitoneally.Two weeks after STZ injection,insulin glargine (92u/day for 8 weeks) was subcutaneously injected in the insulin-treated diabetes group.Then,the rat lumbar spinal cords were collected.The protein levels of phospho-Insulin receptor substrate 1 (P-IRS1),phospho-NMDAR NR1 subunit (P-NR1),P-JNK and P-p44/42MAPK were evaluated by Western blotting.One week before the terminal of the study,paw thermal response latency was measured in all groups.Results Blood glucose levels were tremendously high in both the diabetic group and insulintreated diabetes group.Compared with the control group,paw thermal response latency was markedly shortened in the diabetic group (P＜ 0.001) and the insulin-treated diabetes group (P＜ 0.001),and the alteration was more pronounced in the diabetic group (P＜0.05).Compared with the control group,the protein levels of P-IRS1,P-NR1,P-JNK and P-p44/42MAPK were increased by 79.2％,35.1％,47.6 ％,64.3 ％ and 87.6 ％,respectively in diabetic group and 49.4 ％,19.1％,16.5 ％,31.8％ and 39.9％,respectively in insulin-treated diabetes group (all P＜0.001 or 0.05).In comparison with diabetic group,the increased amplitudes of above 4 parameters were decreased by 29.8％,16.0％,31.2％,32.5％ and 47.7％ respectively in the insulin-treated diabetes group (all P＜0.05).Conclusions NMDAR and its downstream signals,such as JNK and p42/44MARK,are involved in the pathogenesis of painful diabetic neuropathy.Although it could not efficiently control the blood glucose level,low-dose insulin treatment may partly inhibit the occurrence of thermal hyperalgesia through inhibiting NMDAR signal pathway.

Objective To summarize and analyze the shielding,retention and reentry works of blood donors,and to investigate the feasibility of retention and reentry strategy.Methods The samples of ELISA single reagent reactive/NAT non-reactive and ELISA non-reactive/ NAT reactive were negative by confirmatory tests.Then the blood was weeded out and the donation qualification was reserved.The donors of shielding more than 6 months could propose the reentry application at any blood station in the province,and were allowed to return to the ranks after qualified by routine detection and re-detection by Jiangsu Provincial Blood Center.The unqualified rates were compared between the donors of again blood donation after retention and reentry with the common donors by χ2 test.Results From October 2014 to June 2016,1 615 cases were ELISA single reagent reactive/NAT non-reactive,among which 67 cases were confirmed as positive,42 cases were undetermined and 1 506 cases were negative;831 cases were ELISA non-reactive/ NAT reactive,in which 809 cases were positive by confirmation and 22 cases were negative.A total of 1 528 donors were confirmed as negative and their donation qualifications were reserved,89 donors conducted blood donation again and 79 were qualified in blood detection.The unqualified rate was 11.24%,compared with that of common donors,the difference was statistically significant(P<0.001).Meanwhile,596 donors applied for reentry,among them 218 persons were weeded out by the reentry blood station.In remaining 378 samples sent to Jiangsu Provincial Blood Center,359 samples were qualified and confirmed to the reentry condition.Among them,332 donors conducted blood donation and all were qualified by blood detection.Conclusion The reentry strategy in Jiangsu Province is reasonable and feasible,but the donors retention strategy needs to be further optimized and perfected.

Abstract: Objective To clarify the long-term evolution of hepatitis B virus (HBV) quasi-species in HBsAg asymptomatic carriers in Long'an county, Guangxi. Methods ELISA was used to detect serological markers of HBV. Viral loads were measured by real time PCR. HBV DNA was extracted from serum by kits. The whole HBV genome was amplified using nested PCR and amplicons were sequenced by next-generation sequencing (NGS). These sequences from NGS were analyzed by the software like Mega. Results Serum samples were collected from 9 HBsAg asymptomatic carriers in Longan County，Guangxi at 4 different time points in 2004, 2007, 2013, 2019 or 2020. A total of 23 serum samples and 309 full-length gene quasi-species sequences were obtained, with an average amount of (0.18±0.07) G sequencing data for each sample. Genotype of 55.54%(5/9) the studied subjects underwent genotype conversion during the long-term evolution process of HBV quasi-species, and the genotyping results of the phylogenetic tree in the PreS/S region are in perfect agreement with the results of the whole genome analysis; recombinant B/C, I/C were found; the Sn ranged from 0 to 0.37 and the genetic diversity ranged from 0 to 0.11, respectively. A total of 21 special single nucleotide/amino acid mutations (7 in the S region, 2 in the X region, 3 in the PreC region and 9 in the BCP region) and 6 deletion mutations were detected, multiple mutations were found and no drug resistant mutations were found; 77.8%(7/9) of the HBV strains carried by the subjects in 2004 had double mutations at nt1 762(A→T) and 1 764(G→A) and a stop mutation at nt1 896(G→A); HBV mutations can be restored from the mutant type to the wild type and (or) vice versa without antiviral drug pressure, and The evolution rate of HBV genome was 2.03×10-5~3.50×10-3.Conclusion HBV genotype, recombinants, genetic complexity and diversity of HBV quasi-species can change over time during in natural infection. The transformation between HBV mutation type and wild type reduces the value of predicting clinical outcomes by genetic types and related mutations to some extent. The HBV genome evolution rate of asymptomatic carriers of HBsAg in Long'an County is very high.

Objective To compare the survival outcomes between operative versus non-operative treatment of advanced intrahepatic cholangiocarcinoma.Methods This is a retrospective study.The data from 122 patients with intrahepatic cholangiocarcinoma treated at the Hunan People's Hospital,the Hepatobiliary Hospital and the Oncology Department from January 2012 to October 2017 were retrospective studied.87 patients who underwent radical surgery (anatomical hepatectomy + regional lymph node dissection) formed the operation group;35 patients who were treated with chemotherapy and/or radiotherapy and/or biological targeted therapy formed the non-operative group.The general characteristics of the two groups including age,sex,ALT,AST,CA19-9,liver function,Child's classification,AJCC staging,tumor number,vascular (hepatic artery,portal vein) invasion and regional lymph node metastasis rates were compared.The overall survival of the two groups was compared.Results There were no significant differences in age,sex,ALT,AST,CA19-9,liver function,Child's classification,AJCC staging,tumor number,vascular (hepatic artery,portal vein) invasion and regional lymph node metastasis rates (P＞0.05).The overall survival of the operation group was significantly longer than that of the non-operative group (P＜0.05).The mean overall survival for the 2 groups of patients were 32 months and 15 months respectively.The 1-year survival rates were 74.8％ and 58.7％,and the 3 year survival rates were 42.4％ and 6.5％,respectively.The 5 years survival rates were 12.3％ and 0,respectively.Conclusion Operative treatment resulted in better median survival,as well as 1-,3-and 5-year survival rates than non-operative treatment for patients with advanced intrahepatic cholangiocarcinoma.

Objective To evaluate the effect of diammonium glycyrrhizinate extracted from the Chinese traditional medicine licorice root on the growth of human hair follicles cultured in vitro,and to detect the expression of wnt/β-catenin signaling pathway-related molecules.Methods Isolated hair follicles were cultured with diammonium glycyrrhizinate at different concentrations of 0.1,0.01,0.001 and 0.000 1 μmol/L for 10 days,and the hair follicles cultured in Williams' E medium without diammonium glycyrrhizinate served as a control group.The length of hair follicles was measured under a microscope every day,the morphologic changes of hair follicles were observed,and photos were taken.Immunofluorescence assay was performed to assess the proliferation of hair matrix cells,as well as to determine the expression of β-catenin,glycogen synthase kinase 3β (GSK3β),phosphorylated GSK3β (p-GSK3β) and lymphoid enhancer factor-1 (Lef1) in the Wnt/β-catenin signaling pathway.Statistical analysis was carried out by repeated-measures analysis of variance and one-way analysis of variance.Results As repeated-measures analysis of variance showed,only 0.01 μmol/L diammonium glycyrrhetate showed significantly promotive effect on the growth of hair follicles compared with the medium alone (P ＜ 0.05),and there were no significant differences in the length of hair follicles between the other concentration groups and the control group.Compared with the control group,the transition to the catagen phase of human hair cycle was delayed in the 0.01-μmol/L diammonium glycyrrhetate group,while it did not change in the other diammonium glycyrrhetate groups and control group.Immunofluorescence assay showed that the number of ki67-positive hair matrix cells was obviously increased in the 0.1-,0.01-,0.001-μmol/L diammonium glycyrrhizinate groups compared with the control group,while there was no difference between the 0.000 1-μmol/L diammonium glycyrrhizinate group and the control group.One-way analysis of variance revealed that the expression of β-catenin,p-GSK3β and Lef1 significantly differed among all the groups (F =12.604,16.65,15.266 respectively,P ＜ 0.05),while no significant difference in the expression of GSK3β was found among these groups (F =1.472,P ＞ 0.05).Least significant difference (LSD)-t test revealed that the expre-ssion of β-catenin,p-GSK3β and Lef1 in the hair matrix cells was significantly higher in the 0.1-,0.01-,0.001-μmol/L diammonium glycyrrhizinate groups than in the control group (all P ＜ 0.05),but there was no significant difference between the 0.000 1-μmol/L diammonium glycyrrhizinate group and the control group (P ＞ 0.05).Conclusion Diammonium glycyrrhetate at the concentration of 0.01 μmol/L shows markedly promotive effect on the in vitro growth of hair follicles,and can increase the proliferative activity of hair matrix cells and delay the transition to the catagen phase,which may be associated with the activation of Wnt/β-catenin signaling pathway.

Christianson syndrome is a rare X-linked disease caused by mutations in the SLC9A6 gene. The clinical manifestations are male developmental delay, language disorder, seizures, mental retardation, ataxia, microcephaly and so on. Two cases of male children with Christianson syndrome were reported. The proband was 1 year and 11 months old. Clinical manifestations include microcephaly, global developmental delay, and seizures. The electroencephalogram showed that the central midline region of spikes and slow waves were emitted, and all exons sequencing detected a mutation in the SLC9A6 gene chrX: 135084373 [c.803+1(IVS6)G>A]. The proband′s brother was 4 years and 8 months old. The clinical manifestations were similar. The electroencephalogram showed spikes and spines in the Rolandic area on both sides. Slow waves and spiny slow waves were emitted. Magnetic resonance imaging suggested brain atrophy. The genetic verification results were consistent with the proband. The SLC9A6 gene c.803+1(IVS6) G>A splicing mutation was a pathogenic mutation in this family.

Objective To investigate the status quo of pain crisis in advanced lung cancer patients and analyze its influencing factors.Methods From August to November 2023,318 patients with advanced lung cancer were selected from 6 wards of respiratory department of a tertiary A hospital in Zhengzhou.The Numerical Rating Scale,Perceptive Social Support Scale,Self-rating Anxiety Scale and Self-rating Depression Scale were used to investigate the influencing factors of pain crisis in advanced lung cancer patients by Logistic regression.Results Among 318 patients with advanced lung cancer,102 patients had painful crisis,with the incidence rate of 32.08％.0lder age and high level of social support were protective factors for pain crisis,and bone metastasis,anxiety and mild to moderate depression were risk factors for pain crisis.Conclusion The incidence of pain crisis was high in advanced lung cancer patients.Medical staff should pay attention to those with younger age,bone metastasis,low level of social support,high level of anxiety and mild to moderate level of depression,and take timely intervention measures to reduce the occurrence of pain crisis.

Objective The oxidative damage of DNA can be caused by excessive levels of Reactive oxygen species(ROS).Monitoring of DNA oxidative damage enables effective evaluation of ROS damage effects.Although the detection of DNA damage effects based on microbial sensor allows quantitative analysis of oxidative damage,the ROS clearance mechanism existed in bacterial will affect the sensitive of detection.The work of this article is to knockout the key genes of ROS clearance mechanism and construct an antioxidant gene-knock-out microbial sensor.The microbial sensor can realize sensitive monitoring of DNA damage effects and then evaluates the damage effects of cells by ROS.Methods The antioxidant damage genes of bacterial ahpCF,katE and katG were knocked out by λ-Red homologous recombination and antioxidant gene-knockout microbial sensor was constructed.The nalidixic acid sodium salt and UV irradiation were used to characterize the performance for monitoring of DNA damage effects.Results The antioxidant gene-knockout microbial sensors ΔahpC,ΔahpCF/ΔkatEG and ΔahpCF/ΔkatE/ΔkatG were constructed successfully.The results showed that the microbial sensor ΔahpCF/ΔkatE/ΔkatGl had the highest sensitive of damage effects and the limit of detection for nalidixic acid sodium salt was 0.40 μmol/L.In addition,1.80 min of UV irradiation(254 nm)was sufficient to induce a significant fluorescent expression effect in the engineered bacteria.Conclusion In this article,antioxidant gene-knockout microbial sensors had been constructed to realize active and sensitive monitoring of DNA damage effects such as DNA damage reagents and UV irradiation.The sensors could provide an active,effective,and sensitive potential monitoring method for future evaluation of radiation effects in space.

Objective:To investigate the impact of early blood glucose fluctuations after acute multiple injuries on post-traumatic stress disorder (PTSD).Methods:This study was a case-control study. From March 2022 to March 2023, patients with acute multiple injuries who were admitted to the ICU of the Affiliated Hospital of Xuzhou Medical University were selected. According to whether complicated with traumatic brain injury (TBI), the patients were divided into TBI group and non-TBI group. Early post-traumatic blood glucose fluctuations were observed, including stress-induced hyperglycemia (SIH), initial blood glucose value on admission, blood glucose extreme, short-term glycemic variability (GV) and other related indicators. The 72-hour glucose coefficient of variation (Glu-CV) was used to reflect short-term GV. After 1 month, the PTSD checklist for DSM-5 (PCL-5) was used to assess the patient's symptoms of PTSD. The patients were divided into PTSD group and non-PTSD group according to PCL-5 score ≥38. The differences in short-term glucose fluctuations in each groups were compared; the risk factors of PTSD were analyzed by logistic regression; the receiver operating characteristic (ROC) curve was plotted to evaluate the predictive value of related indicators on the incidence of PTSD.Results:159 patients with acute multiple injuries were selected and defined as the TBI group ( n=94) and non-TBI group ( n=65). The incidence of PTSD, PCL-5 scale scores, the incidence of SIH and 72 h Glu-CV in the TBI group were significantly higher than the non-TBI group (all P<0.05). The incidence of SIH and 72 h Glu-CV in the PTSD group were significantly higher than the non-PTSD group (both P<0.05). Multivariate logistic regression analysis showed that 72 h Glu-CV ( OR=1.333, 95% CI: 1.028-1.727, P=0.030) was the independent risk factor for PTSD after acute multiple injuries, and the area under the ROC curve was 0.861 (95% CI: 0.789-0.933, P<0.001), the sensitivity was 62.9% and the specificity was 93.5%. Conclusion:Patients with acute multiple injuries with TBI are more likely to have early glucose fluctuations and develop PTSD, and increased short-term glucose variability is the independent risk factor for PTSD after acute multiple injuries.