Objective This study aimed to identify PAX9 variants in non-syndromic tooth agenesis families of Chi-na,as well as to analyze the genotype-phenotype of non-syndromic tooth agenesis caused by PAX9 variants,which can provide a basis for the genetic diagnosis of tooth agenesis.Methods We collected the data of 44 patients with non-syn-dromic oligodontia who underwent treatment at Stomatological Hospital of Hebei Medical University between 2018 and 2023.Whole-exome sequencing was performed on the peripheral blood of the proband and its core family members,and the variants were verified by Sanger sequencing.Pathogenicity analysis and function prediction of the variants were per-formed using bioinformatics tools.The correlation between the genotype of PAX9 variant and its corresponding pheno-type was examined by reviewing 55 publications retrieved from PubMed.The studies involved 232 tooth agenesis pa-tients with PAX9 variants.Results A novel PAX9 c.447delG(p.Pro150Argfs*62)and a reported PAX9 c.406C>T(p.Gln136*)were identified in two Chinese families.Through bioinformatics analysis and three-dimensional structural mod-eling,we postulated that the frameshift variant was pathogenic.The outcome was the premature cessation of PAX9 pro-tein,which caused severe structural and functional deficiencies.Summarizing the PAX9 genotype-phenotype relationship revealed that patients carrying the PAX9 variant commonly led to loss of the second molars.Conclusion We identified the novel PAX9 c.447delG(p.Pro150Argfs*62)in a Chinese family of non-syndromic oligodontia,expanding the known variant spectrum of PAX9.The most susceptible tooth position for PAX9 variants of tooth agenesis was the second mo-lars and the deciduous molars during the deciduous dentition.

Objective To investigate the protective effect and molecular mechanism of bloodletting acupuncture at Jing-well points on acute high-altitude hypoxia brain injury through regulating PI3K/AKT/mTOR signaling pathway-mediated mitochondrial autophagy,and to provide an effective target and theoretical basis for the clinical use of bloodletting acupuncture at Jing-well points to prevent and treat acute high-altitude hypoxia brain injury.Methods Male SD rats were randomly divided into Control group(n=15)and experimental group,and the experimental group was divided into Model group(n=15),Bloodletting Acupuncture at Jing-well Points of hand group(BAJP group,n=15),Bloodletting Acupuncture at Non-Acupoint group(BANA group,n=15).The low pressure oxygen chamber was depressurized to 6000 m altitude,and the rats in each experimental group were treated with low-pressure hypoxia for 72 h to replicate the acute high-altitude hypoxia brain injury rat model.The rats in the BAJP group were bled according to the order of"Shaoshang"(LU11),"Shangyang"(LI1),"Zhongchong"(PC9),"Guanchong"(SJ1),"Shaochong"(HT9),"Shaoze"(SI1),once a day for 7 days.The rats in the BANA group were bled by cutting the tail tip daily,and the amount of blood bled was 15-20 μL in both groups.The expression levels of PI3K,AKT and mTOR in hippocampal tissues of rats were detected by Western blot;AKT and mTOR mRNA expression levels in hippocampal tissues of rats were detected by PCR.Results Compared with the Control group,the number of degenerative necrotic vertebral cells in CA1 area of hippocampal tissue,swelling of mitochondria,appearance of autophagosomes,and increase of apoptosis in hippocampal tissue of Acute High-altitude Hypoxia(AHH)rats were significantly increased;After bloodletting acupuncture at Jing-well points of hand treatment,various brain injury manifestations in AHH rats were alleviated;Bloodletting acupuncture at non-acupoint had no significant ameliorating effect on AHH rats′ brain injury.Western blot detected a significant decrease in the phosphorylation levels of PI3K,AKT,and mTOR in the hippocampal tissues of AHH rats compared to Control group rats(P<0.01),and the phosphorylation levels of the three molecules were further decreased after bloodletting acupuncture at Jing-well points of hand treatment(P<0.01),and bloodletting acupuncture at non-acupoint treatment did not have significantly affect on the phosphorylation levels of these molecules(P>0.05),and AKT,mTOR mRNA expression levels further demonstrated the above trend.Conclusion Bloodletting acupuncture at Jing-well points of hand can play a protective role against acute high-altitude hypoxia brain injury with points specificity,and the mechanism may be related to the inhibition of PI3K/AKT/mTOR pathway to promote the elevated level of mitochondrial autophagy,improve mitochondrial physiology,and enhance the body′s ability to resist apoptosis and hypoxia.

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

Objective:To explore the clinical characteristics and establish a corresponding prognostic scoring model in patients with early-stage clinical features of hepatitis B-induced acute-on-chronic liver failure (HBV-ACLF).Methods:Clinical characteristics of 725 cases with hepatitis B-related acute-on-chronic hepatic dysfunction (HBV-ACHD) were retrospectively analyzed using Chinese group on the study of severe hepatitis B (COSSH). The independent risk factors associated with 90-day prognosis to establish a prognostic scoring model was analyzed by multivariate Cox regression, and was validated by 500 internal and 390 external HBV-ACHD patients.Results:Among 725 cases with HBV-ACHD, 76.8% were male, 96.8% had cirrhosis base,66.5% had complications of ascites, 4.1% had coagulation failure in respect to organ failure, and 9.2% had 90-day mortality rate. Multivariate Cox regression analysis showed that TBil, WBC and ALP were the best predictors of 90-day mortality rate in HBV-ACHD patients. The established scoring model was COSS-HACHADs = 0.75 × ln(WBC) + 0.57 × ln(TBil)-0.94 × ln(ALP) +10. The area under the receiver operating characteristic curve (AUROC) of subjects was significantly higher than MELD, MELD-Na, CTP and CLIF-C ADs( P < 0.05). An analysis of 500 and 390 cases of internal random selection group and external group had similar verified results. Conclusion:HBV-ACHD patients are a group of people with decompensated cirrhosis combined with small number of organ failure, and the 90-day mortality rate is 9.2%. COSSH-ACHDs have a higher predictive effect on HBV-ACHD patients' 90-day prognosis, and thus provide evidence-based medicine for early clinical diagnosis and treatment.

Objective To understand the distribution of water iodine in the external environment of Shanxi Province, and to provide a basis for redefining and implementation of scientific iodine supplementation in iodine deficiency, iodine adequate or iodine high areas. Methods In 2012 - 2016, administrative villages (neighborhood committees)in 119 counties(cities,districts)in 11 cities of Shanxi Province were selected as the investigation units, and 1 to 5 drinking water samples were collected. Water iodine content was detected using arsenic-cerium catalytic spectrophotometry. Areas standard was designated: water iodine content < 10 μg/L as iodine deficiency areas, > 100 μg/L as iodine high areas. Results A total of 26 213 administrative villages (neighborhood committees) of 1 362 townships (towns) of 119 counties (cities, districts) were surveyed, covering 2 850.94 ten thousand people. A total of 32 766 water samples were collected and the median iodine was 5.2 μg/L. There were 18 199 villages with water iodine < 10 μg/L, accounting for 69.4% (covering 1 812.17 ten thousand people, accounting for 63.6%), 6 471 villages with water iodine 10-<50 μg/L,accounting for 24.7%(covering 787.44 ten thousand people,accounting for 27.6 %),1 166 villages with water iodine 50 - < 100 μg/L,accounting for 4.4% (covering 181.46 ten thousand people, accounting for 6.4%), 377 villages with water iodine ≥100 μg/L, accounting for 1.4%(covering 69.87 ten thousand people, accounting for 2.5%).In 1 362 townships (towns), 71.1% (969)water iodine median was<10 μg/L,24.2%(330)water iodine median was in 10-<50 μg/L,3.4%(46)water iodine median was in 50-<100 μg/L,and 1.2%(17) water iodine median was ≥100 μg/L. In 119 counties(cities,districts),there were 90 counties(accounting for 75.6%) with the water median < 10 μg/L, there were 26 counties (accounting for 21.8%) with the water median 10 - < 50 μg/L. Conclusions Most parts of Shanxi Province(or the resident population) are iodine deficiency areas, the external environment water iodine contents in the rest of the regions are different, we should adopt different iodine supplement or iodine reduction measures in regions with different water iodine levels.

Objective To study the nutritional status of pregnant women in Shanxi Province before and after the implementation of the new standards of iodized salt content,provide the basis for scientific supplementation of iodine for pregnant women.Methods According to the method of population proportion sampling,30 county-level monitoring sites were selected,a primary school was selected from each county (city,district) by the method of simple random sampling and 40 students in 2011 or 50 students in 2014 aged 8-10 years were selected in each school,direct titration was used to detect salt iodine;at the same time,20 pregnant women were selected from each town where the primary school was located and urinary iodine was determined using arsenic cerium catalytic spectrophotometry (WS/T 107-2006).Results A total of 1 182 and 1 437 salt samples was detected in Shanxi Province in 2011 and 2014,the median of salt iodine was 30.5 and 24.1 mg/kg,respectively,and the difference was statistically significant (H =567.45,P ＜ 0.01);it was 95.41％,80.31％,76.62％ of the coverage rate of iodized salt,qualified rate of iodized salt,qualified iodized salt consumption rate in 2014,respectively;which were compared with those in 2011 (97.63％,97.49％,95.18％),the differences were statistically significant (x2 =9.27,232.40,166.25,P ＜ 0.01).A total of 440 and 630 urinary samples of pregnant women were tested in 2011 and 2014,the median of urinary iodine was 279.6 and 177.1 μg/L,respectively,iodine nutrition of pregnant women was more than adequate in 2011,and iodine nutrition was suitable in 2014.The difference was statistically significant (H =153.89,P ＜ 0.01).The proportion of pregnant women's median of urinary iodine less than 150 μg/L in 2014 [41.11％ (259/ 630)] was significantly higher than that in 2011 [8.18％ (36/440),x2 =140.68,P ＜ 0.01].The constituent ratio of 250 to 500 μg/L was significantly decreased [23.65％ (149/630) vs 54.77％ (241/440),x2 =108.33,P ＜ 0.01).Conclusion It is at a reasonable level of iodine nutrition level of pregnant women in Shanxi after the adjustment of iodized salt content,but the ratio of ＜ 150 μg/L is increasing,which needs to be paid attention to.

Objective To investigate the tumor/ non-tumor (T/ NT) ratio during 99 Tcm-hydrazinon-icotinamide(HYNIC)-( poly-( ethylene glycol), PEG) 4-Glu( cyclo ( Arg-Gly-Asp-D-Phe-Lys ( PEG4 ))) 2 ( 99 Tcm-3PRGD2 ) SPECT/ CT imaging and clinical pathological features of breast cancer. Methods Forty-five female patients (age range: 39-76 (53.0±9.5) years) with suspected breast malignant nodules or mas-ses from October 2016 to June 2017 were prospectively enrolled. Patients underwent 99 Tcm-3PRGD2 SPECT/CT imaging before breast puncture and surgery. All subjects had pathological results, and estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER-2), Ki-67 and mi-crovessel density ( MVD) were obtained by immunohistochemistry and fluorescence in situ hybridization(FISH). Two-sample t test and Kruskal-Wallis H test were used to analyze the data. Results Invasive duc-tal carcinoma was pathologically confirmed in 35 of 45 patients. There were 7 patients in stage Ⅰ, 11 pa-tients in stage ⅡA and 17 patients in stage ⅡB. The Luminal A subtype, Luminal B subtype, ERBB2+subtype, Basal-like subtype were found in 6, 9, 9 and 11 patients, respectively. The T/ NT ratio was signifi-cantly higher in the stageⅡB patients than that in stageⅠ+ⅡA patients (4.54±1.46 vs 3.32±1.72, t= -2.24, P<0.05). Patients with ERBB2+ subtype had higher T/ NT ratio compared to patients with Basal-like sub-type: 5.80(3.90, 6.70) vs 2.80(2.20,3.50), H= 11.06, χ2 = 15.31, both P<0.05. Besides, the T/ NT ra-tios in the HER-2 positive group and lymphatic metastasis group were significantly higher than those in the HER-2 negative group and group without lymph node metastasis (t values: -3.99, -2.51, both P<0.05). MVD of HER-2 positive group was higher than that of HER-2 negative group (t= 7.13, P<0.01). Conclu-sion The T/ NT ratio during 99 Tcm-3PRGD2 SPECT/ CT imaging has relations with TNM staging, lymph node infiltration and HER-2 in breast cancer.

To confirm the etiology of a dead case for a 6 year-old female Ailurus fulgens,one strain of the predominant bacteria from pathologic tissues(heart,liver,spleen,lung and other samples) of the dead Ailurus fulgens were examined and isolated.The isolate was named R1 and no other bacteria were isolated.The bacterial etiological examination(morphological characteristics,biochemical characteristics and 16S rDNA gene detection)of R1 showed that it was identifed as K.peneumoniae.Artificial infection to mice about R1 was also conducted in this study.R1 had strong pathogenicity to mice and the LD50 is 6.5 × 104 CFU/mL.Moreover,the clinical and pathological features of the dead mice were consistent with that of the Ailurus fulgens.To find effective therapeutic drugs of curing other Ailurus fulgens,antibiotic sensitivity test of R1 was conducted,and the results revealed that R1 was highly sensitive to cefotaxime et al,moderately sensitive to amikacin and resistant to penicillin.These data showed that K.peneumoniae was bacterial pathogen leading to death of the Ailurus fulgens and it had strong resistance to penicillins,macrolides and virginiamycin and it had broad drug resistance spectrum.However,R1 is sensitive to cephalosporins and aminoglycoside antibiotics.

Objective To investigate resident iodine nutrition level in waterborne high iodine areas and prevalence of high iodine goiter in Shanxi Province.Methods In 2014,in Shanxi Province,in all the 10 high iodine counties (cities,districts),the jurisdiction area of each county (city,district) was divided into two blocks,high iodine and not high iodine districts,and in high iodine area of each county (city,area) according to their sub-area positions of east,south,west,north and center,a township was randomly selected (if the number of high iodine area in iodine excessive township ≤5,all townships were selected);four administrative villages in each monitoring township were randomly selected;in each administrative village,the edible salt samples of 15 randomly selected households were collected for detection of iodine content.Five counties (cities,districts) were selected from the province's 10 high iodine counties (cities,districts),water iodine content of 3 counties (cities,districts) was 150-300 μg/L,and 2 were ＞ 300 μg/L,one administrative village was selected from each county (city,area),household drinking water samples were collected to detect iodine content;and 100 elementary school children aged 8-10 were selected from the village where the monitoring stations located in for thyroid volume ultrasound measurement,and 30 of them were randomly selected for urinary iodine content detection.Results ① In the 10 high iodine counties (cities,districts),1 680 households salt samples were detected,and the rate of no iodine salt samples was 89.2％(1 499/1 680).② Fourteen water samples were collected in 5 counties (cities,districts),and the water iodine content was 155.7-467.3 μg/L.③ In the five high iodine counties (cities,districts),197 urine samples were collected from children aged 8 to 10.The median of urinary iodine was 466.5 μg/L;the median urinary iodine was 650.1 μg/L in water iodine≥300 μg/L children group which was significantly higher than that of the group with 150-300 μg/L water iodine content 332.5 μg/L (Z =-6.164,P ＜ 0.05);urinary iodine level of children and the water iodine of the corresponding village was positively correlated (r =0.543,P ＜ 0.05).④ In the five high iodine counties (cities,districts),543 children aged 8-10 were measured with their thyroid volume,the thyroid goiter rate was 6.8％(37/543);the goiter rate of water iodine ≥300 μg/L children group was [9.0％(28/311)] which was significantly higher than that in the iodine content of drinking water 150-300 μg/L group [3.9％(9/232),x2 =5.494,P ＜ 0.05].Conclusion The measurement of stopping iodized salt supply in high iodine areas in Shanxi Province is well implemented,iodine nutrition level and thyroid goiter rate in those areas are still too high,high iodine intervention measures can be focused on changing of the drinking water.

Objective To master the iodine nutritional status of residents and to evaluate the effect of iodine nutrition improvement on residents health after the adjustment of salt iodine concentration. Methods According to the method of population proportion sampling, 30 county-level monitoring sites were selected in 2014, a primary school was selected from each county (city, district) by the method of simple random sampling and 50 students aged 8 - 10 (half males and half females) were selected in each school, B ultrasound was used to detect thyroid volume, arsenic cerium catalytic spectrophotometry (WS/T 107-2006) was used to detect urinary iodine, direct titration was used to detect salt iodine; At the same time, 20 pregnant women were selected from each town and urinary iodine was determined. All results were compared with the results of 2011 to evaluate the effect of iodine nutrition improvement on resident's health after the adjustment of salt iodine concentration. Results A total of 1 437 edible salt samples were tested, the median of salt iodine concentration was 24.1 mg/kg. Based on the new standard (18 - 33 mg/kg), the coverage rate of iodized salt was 95.4%(1 371/1 437), the qualified rate of iodized salt was 80.3%(1 101/1 371), the consumption rate of qualified iodized salt was 76.6% (1 101/1 437). A total of 1 496 urine samples of children and 630 urine samples of pregnant women were tested, the median of urinary iodine of children and pregnant women was 224.6 and 177.1 μg/L, respectively, which were significantly lower than those of 2011 (274.6, 279.6μg/L), the differences were statistically significant (H=70.10, 153.50, all P<0.05). The proportion of counties which iodine nutrition of children and pregnant women in suitable range was 36.7% (11/30) and 56.7% (17/30), which were higher than those of 2011 [6.5%(2/31), 25.8%(8/31)], the differences were statistically significant (χ2 = 7.88, 5.00, all P < 0.05). Totally 1 552 children were examined thyroid in Shanxi Province, the rate of goiter was 4.4% (69/1 552). Conclusion The new standard of iodine concentration makes it possible to maintain a sustainable elimination of iodine deficiency disorders, and it can improve the iodine nutrition of children aged 8-10 years and pregnant women to more reasonable level.