Objective:To establish an HPLC characteristic chromatogram method to simultaneously determine the content of 6 components in Bombyx Batryticatus and its processed products; To provide a scientific means for quality evaluation.Methods:The HPLC analysis was performed on a Waters Xselect HSS T3 column (250 mm×4.6 mm, 5 μm), with a mobile phase consisting of acetonitrile and 5 mmol/L ammonium acetate solution (adjust pH to 4.5 with acetic acid) in gradient mode at a flow rate of 1.0 ml/min. The column temperature was 30 ℃. The detection wavelength was 260 nm. The injection volume was 10 μl. The SIMCA-P 14.0 software was used for cluster analysis and partial least squares discriminant analysis to evaluate the quality of Bombyx Batryticatus and its processed products.Results:Nine common peaks were identified in the HPLC characteristic spectra of Bombyx Batryticatus and its processed products, and seven common peaks were identified. Cluster analysis and partial least squares discriminant analysis can classify Bombyx Batryticatus and its processed products into two groups, with peak 4, peak 2, peak 1, peak 6, and peak 3 as their mass difference markers. Uric Acid, Hypoxanthine, Uridine, Adenine, Guanosine, and Adenosine showed good linearity ( R2≥0.999 9) within 43.95-879.04, 2.05-41.07, 2.35-47.09, 2.02-40.36, 2.32-46.43, 2.32-46.43 μg/ml, respectively. The precision, stability (24 h), repeatability relative standard deviation (RSD) were all <3.00%. The average spiked recoveries were all in the range of 94.57%-102.01%, and the RSDs were all in the range of 0.96%-3.32%. In 15 batches of Bombyx Batryticatus, the content of the above 6 components, was determined to be 11.51, 0.33, 0.38, 0.42, 0.35, 0.36 mg/g in the following order. And in 15 batches of fried Bombyx Batryticatus, the content of components was determined to be 12.06, 0.35, 0.36, 0.18, 0.21, 0.24 mg/g in the following order. Conclusions:The method established in this study is simple, rapid, and repeatable. PLS-DA can distinguish Bombyx Batryticatus and its processed products, which can provide a reference for the quality evaluation of Bombyx Batryticatus and its processed products.

Objective:To establish a mass spectrometry method for the determination of characteristic peptides of Galli Gigerii Endothelium Corneum that can identify the authenticity of Galli Gigerii Endothelium Corneum as well as its preparations; To evaluate their quality.Methods:Ultra performance chromatography-electrospray tandem mass spectrometry (UPLC-MS/MS) with the mode of multiple reaction monitoring quantification (MRM) was employed to monitor the ion pairs of m/z 379.21(charge: +2)→571.36, m/z 379.21(charge: +2)→385.26, m/z 785.41(charge: +2)→941.51 and m/z 785.41(charge: +2)→245.08, in order to detect the Galli Gigerii Endothelium Corneum crude drug and its preparations. Results:Chicken specific peptide I and chicken specific peptide Ⅱ could be detected in the 18 batches of Galli Gigerii Endothelium Corneum from different regions, their corresponding extractions and 4 batches of prescription preparations, while the chicken specific peptides were not detected in the 8 batches of endothelium corneums from ducks, geese and pigeons.Conclusions:The method established in this study can effectively supplement the deficiencies in standards of Galli Gigerii Endothelium Corneum and its decoction pieces, improve the quality control standard, and provide a reference for the safety and effectiveness of Galli Gigerii Endothelium Corneum in clinical medication.

Objective:To investigate the prevalence of adult skeletal fluorosis caused by drinking tea-type endemic fluorosis in Yushu Tibetan Autonomous Prefecture (hereinafter referred to as Yushu Prefecture), Qinghai Province, and provide scientific basis for prevention and control of the disease.Methods:In August 2021, one village was selected as a survey site in six counties (cities) in Yushu Prefecture, including Nangqian, Chindu, Yushu, Zadoi, Qumarlêb, and Zhiduo. Drinking water samples and 10 brick tea samples were collected from each village to determine the fluoride content in water and brick tea; at least 100 permanent residents aged ≥ 25, who had a habit of drinking brick tea and had lived in the local area for more than 5 years, were selected for X-ray imaging to examine the prevalence of adult skeletal fluorosis.Results:A total of 75 samples of residential drinking water were collected, with a fluoride content of (0.21 ± 0.05) mg/L, ranging from 0.11 to 0.34 mg/L; 60 samples of brick tea, with a fluoride content of (626.70 ± 157.27) mg/kg, ranging from 324.00 to 2 102.00 mg/kg. A total of 1 136 adults were examined, and 318 cases of skeletal fluorosis were diagnosed, with a detection rate of 27.99%. Among them, the detection rates of mild, moderate, and severe skeletal fluorosis were 20.95% (238/1 136), 6.07% (69/1 136), and 0.97% (11/1 136), respectively, with mild symptoms being the main. The detection rates of skeletal fluorosis in males and females were 29.09% (121/416) and 27.36% (197/720), respectively, with no statistically significant difference between the gender (χ 2 = 0.39, P = 0.533). Comparison of the skeletal fluorosis in different gender, the differences were statistically significant (χ 2 = 22.31, P < 0.001). The detection rates of skeletal fluorosis in the age groups of 25 - 35, 36 - 45, 46 - 55, 56 - 65, 66 - 75, and ≥76 years old were 6.86% (7/102), 22.37% (51/228), 24.02% (92/383), 37.44% (73/195), 43.48% (70/161), and 37.31% (25/67), respectively. The differences between the groups were statistically significant (χ 2 = 59.84, P < 0.001). Moreover, there was a statistically significant difference in the composition of skeletal fluorosis among different age groups ( H = 37.66, P < 0.001). The Spearman correlation analysis results showed that the severity of adult skeletal fluorosis was positively correlated with age ( r = 0.34, P < 0.001). Conclusions:There is a certain degree of prevalence of adult skeletal fluorosis in Yushu Prefecture. And as age increases, the condition of skeletal fluorosis becomes more severe.

[Objective]To explore if the enhanced recovery after surgery (ERAS) protocol for pregnant women with gestational diabetes mellitus (GDM) who are undergoing elective cesarean delivery could cause perioperative glycemic abnormalities and heighten the risk of neonatal hypoglycemia.[Methods]A retrospective analysis was conducted on a cohort of pregnant women with singleton pregnancies who underwent elective cesarean sections and received ERAS between May 1,2022,and October 31,2023,at the Third Affiliated Hospital of Sun Yat-sen University. A total of 150 patients were included in this study,comprising the GDM group (n=75) and the non-GDM group (n=75). The study included pregnant women with good glycemic control (GDM) and maternal age (18-30 years;30-35 years;35-40 years;>40 years),BMI (<18.5 kg/m2;18.5-24.9 kg/m2;25-30 kg/m2;>30 kg/m2),and gestational age (within 7 days). We used these criteria to match 1∶1 non-GDM women as the control group. After administering preoperative oral carbohydrates,we observed the trends of maternal glycemic changes,including hyperglycemia and hypoglycemia,at any time of the day. We also evaluated the incidence of hypoglycemic low Apgar scores in newborns,abnormal pH values in blood gases,and the rate of transfer to the pediatric unit immediately after delivery.[Results]No significant difference was observed in fasting blood glucose levels on the day of surgery between the two groups of pregnant women[(4.4±0.5) mmol/L vs. (4.3±0.5) mmol/L,t=1.395,P=0.165]. The blood glucose peak was reached 30 minutes after consuming 300 mL (42.6 g of low-dose carbohydrate) of a light drink[(7.2±0.9) mmol/L vs. (6.4±0.8) mmol/L,t=5.773,P<0.001],with a subsequent decline in blood glucose levels. At the 120-minute mark,blood glucose had returned to the pre-oral carbohydrate level. The blood glucose levels in GDM groups was significantly higher than those in the non-GDM group (P<0.005). Although the incidence of hyperglycemia was significantly higher in the GDM group than in the non-GDM group at the 30-minute peak blood glucose level after oral carbohydrate intake,and the difference was statistically significant (17.3％ vs. 1.3％,x2=11.354,P<0.001),severe hyperglycemia (≥10 mmol/L) did not occur. The incidence of hypoglycemia was not significantly higher in neonates in the GDM group than in the non-GDM group (22.7％ vs. 28％,x2=0.564,P=0.453). The incidence of neonatal hypoglycemia in the GDM group was not significantly elevated in comparison to the non-GDM group after adjusting for age and BMI (Model 1),primiparity and gestational week of delivery (Model 2),hypertensive disorders of pregnancy (Model 3),cesarean section indications,time of cesarean section,and intraoperative hemorrhage (Model 4),and neonatal weight (Model 5).[Conclusion]In GDM patients with excellent glycemic control,an ERAS regimen with a low oral dose of carbohydrates prior to elective cesarean section does not increase the risk of preoperative serious hyperglycemia in mothers,nor does it increase the incidence of neonatal hypoglycemia.

Objective To clone Rhipicephalus linnaei Boophilin-H2 gene, construct the recombinant expression vector, express the Boophilin-H2 recombinant protein in Escherichia coli, and assess its anticoagulant activity. Methods Specific primers were designed to amplify the Boophilin-H2 gene fragment using cDNA, synthesized from engorged Rhipicephalus linnaei tick RNA through reverse transcription, as a template. The gene fragment was cloned and connected to plasmid pSmart-I, and the recombinant expression vector pSmart-I/Boophilin-H2 was constructed. The recombinant expression vector was verified by double restriction enzyme digestion with BamHⅠand XhoⅠ, transferred into the competent state of Escherichia coli BL21(DE3, and expressed under low-temperature induction with IPTG. The recombinant protein was purified by Ni-NTA Resin, and its expression and purification were detected by 12.5% SDS-PAGE. The femoral venous blood of New Zealand white rabbits was collected by 3.8% sodium citrate blood collection tube, and the upper plasma was centrifugally separated to measure the anticoagulant activity of the recombinant protein using four test plates of in vitro coagulation. Results A 387 bp gene fragment of Boophilin-H2 of Rhipicephalus linnaei was successfully amplified and cloned; the prokaryotic expression vector pSmart-I/Boophilin-H2 was constructed and verified by double enzyme digestion. Following induction with 0.8 mmol/L IPTG for 16 hours in Escherichia coli, SDS-PAGE showed that the recombinant protein was expressed in the supernatant primarily in a soluble form, with the Boophilin-H2 recombinant protein approximately 35 000 in size. The anticoagulant activity assays of the purified recombinant protein Boophilin-H2 showed that the recombinant protein significantly prolongs the activated partial thromboplastin time (APTT) in a concentration-dependent manner, while its effects on thrombin time (TT), prothrombin time (PT), and fibrinogen (FIB) levels were not significant. Conclusions The expression vector for Boophilin-H2 was successfully constructed, and its product exhibited potent APTT anticoagulant activity, implying its role in the intrinsic coagulation pathway, possibly acting upon intrinsic coagulation factors VIII, XI, and XII to inhibit blood coagulation. This study provides a reference and theoretical foundation for further research and development of tick control vaccines and anticoagulant drugs.

Objective:To learn about the detection quality and external quality control assessment of fluoride and arsenic in laboratories at all levels in Qinghai Province.Methods:The Z-score method was used to analyze and evaluate the evaluation results of 1 provincial, 8 municipal and 43 county level laboratories of disease prevention and control institutions participating in the external quality control assessment of water fluoride and brick tea fluoride in Qinghai Province in 2021, as well as 1 provincial, 1 municipal and 2 county level laboratories of disease prevention and control institutions participating in the external quality control assessment of water arsenic and urine arsenic. The feedback rate and qualification rate of external quality control of each assessment laboratory were calculated.Results:In 2021, the feedback rate of external quality control of water fluoride, brick tea fluoride, water arsenic and urine arsenic in provincial and municipal level laboratories of Qinghai Province were 100.00%; except that the qualified rate of water fluoride was 7/9, the qualified rate of external quality control of other projects was 100.00%. The feedback rate of external quality control of water fluoride, brick tea fluoride, water arsenic and urine arsenic in county level laboratories was 100.00%; except that the qualified rate of water fluoride was 86.05% (37/43), the qualified rate of external quality control of other projects was 100.00%. In the specific assessment results of the laboratory, the assessment results of water fluoride sample FS20210101 from 1 provincial, 1 municipal and 2 county level laboratories, and FS20210102 from 1 county level laboratory were suspicious; the assessment results of water fluoride sample FS20210101 from 3 county level laboratories were not satisfactory; the assessment results of fluoride and arsenic sample in other laboratories were satisfactory.Conclusions:The qualified rate of external quality control of fluoride and arsenic in laboratories at all levels in Qinghai Province is relatively high, but some county level laboratories are still dissatisfied with the assessment results of water fluoride. Therefore, it is necessary to strengthen the detection level of water fluoride in laboratories.

Objective:To follow up the iodine nutrition and intellectual development of school children aged 8-10 years old in Nangqian County of Qinghai Province after 9 years of the implementation of iodine deficiency disorders intervention measures with iodine supplement as the main measure.Methods:In order to improve the iodine nutritional status of the population in Nangqian County, free iodized salt distribution, health education and other intervention measures for iodine deficiency disorders were implemented for 9 consecutive years since 2013. In May 2012 (before iodine supplementation) and September 2021 (after iodine supplementation), the same 5 townships (towns) were selected, and children aged 8-10 years old (half male and half female, age balanced) born locally in the central primary school of each township (town) were selected as the survey subjects. Household edible salt samples and random urine samples were collected for salt iodine and urinary iodine detection; the second revised version of the Chinese Combined Raven's Test (rural version) was used to assess the intelligence quotient (IQ) of children. The Flynn effect (FE) gain was used to adjust IQ, the corrected IQ = (IQ starting point value - current year's IQ value) - FE gain (calculated by 0.74/year). The differences of salt iodine and urinary iodine related indexes, IQ value and corrected IQ were compared before and after iodine supplementation.Results:After iodine supplementation, the coverage rate of iodized salt reached 100.00% (300/300), and the consumption rate of qualified iodized salt reached95.00% (285/300), the median urinary iodine increased to 157.20 μg/L, and all indexes met the elimination standard of iodine deficiency disorders. The IQ value of children aged 8-10 years old after iodine supplementation was 99.00 ± 14.90, significantly higher than that before iodine supplementation (82.00 ± 13.20, F = 156.82, P < 0.001). The FE gain in 9 years was 6.66, and the actual IQ gain of children aged 8-10 years old after iodine supplementation was 10.34. There were statistically significant differences in IQ value before and after iodine supplementation in male and female children ( F = 78.84, 78.88, P < 0.001). After iodine supplementation, there was a statistically significant difference in IQ value between children in the 8-year-old group and the 10-year-old group ( P = 0.010). There were statistically significant differences in IQ value before and after iodine supplementation in 8, 9 and 10 years old groups ( F = 55.23, 65.79, 36.85, P < 0.001). Conclusion:Intervention measures for iodine deficiency disorders, mainly iodine supplement, can significantly improve the iodine nutrition status of children aged 8-10 years old, and significantly promote the intellectual development of children.

Objective:To analyze the external quality control assessment results of urinary iodine, salt iodine and water iodine in iodine deficiency disorders laboratories in Qinghai Province, to evaluate the testing capacity of provincial, municipal and county-level laboratories and the operation of external quality control network, so as to provide quality assurance for consolidating and eliminating iodine deficiency disorders.Methods:In 2021, 1 provincial, 8 municipal, and 43 county-level laboratories in Qinghai Province had participated in the assessment of urinary iodine and salt iodine, while 1 provincial and 8 municipal-level laboratories had participated in the assessment of water iodine. The assessment results were evaluated using the method of reference value ± uncertainty of external quality control samples.Results:All laboratories that participated in the assessment had provided feedback. One provincial-level laboratory passed the assessment of urinary iodine, salt iodine, and water iodine. Among 8 municipal-level laboratories, 2 laboratories failed the urinary iodine assessment, with a pass rate of 6/8; the assessment of salt iodine and water iodine in 8 laboratories were all qualified. Among 43 county-level laboratories, 7 laboratories failed the urinary iodine assessment, with a pass rate of 83.7% (36/43); the assessment of salt iodine in 43 laboratories were all qualified.Conclusions:The external quality control network of iodine deficiency disorders laboratories in Qinghai Province has fully covered all municipal and county-level laboratories. The testing capability of provincial-level laboratory is stable and maintains a high level; the testing quality of some municipal and county-level laboratories is still unstable and needs to be further strengthened.

Abstract: Objective To understand the morphological characteristics and ultrastructure of the dominant species of Rhipicephalus sanguineus in Hainan at different developmental stages, and provide theoretical basis for the identification of the lineage and control of Rhipicephalus sanguineus. Methods The external morphology of different developmental stages of the dominant species of Rhipicephalus sanguineus, including larva, nymph and adult tick in Hainan were observed by scanning electron microscope. Results The division between each segment of larva pedipalps was not obvious, and setae was serrated; dental formula type 2 | 2; 3 pairs of podomere; a pair of setae on the anal valve; none of anal groove, spiracular plate, porous area and genital aperture. There was a clear boundary at the beginning of each segment of nymph pedipalps; dental formula type 2 | 2; 4 pairs of podomere; 3 pairs of setae on the anal valve; anal groove; none of porous area and genital aperture. The male adult tick's trichotheca are covered by the pedipalps, and the whole bristles are conical; dental formula type 3 | 3; 4 pairs of podomere; anal groove and paraprocts; 7 setae on the anal valve; genital aperture was oval. The female of adult tick can be distinguished by dental formula 3 | 3; pairs of podomere; porous areas with 3 short setae; anal groove; 4 pairs of setae and 2 pores on the anal valve; genital pore was broadly U-shaped. In addition, the male adult's scutum occupies almost the entire dorsal surface, the basis capituli of larva, nymph and adult tick all were hexagonal, and the existence of Haller's organ was found on the first pair of legs. Conclusions Scanning electron microscopy observation of the different developmental stages of R.sanguineus revealed clear morphological features, preliminarily suggesting that R.sanguineus in Hainan Province may belong to the tropical lineage, which provide a certain experimental basis for the identification of the tick and the comprehensive prevention and control of local tick-borne diseases.

Objective:To evaluate the water fluoride detection ability of county (city, district) level (referred to as the county-level) laboratories in Qinghai Province.Methods:During the "13th Five-Year Plan" period (2016 - 2020), 4, 4, 4, 43, 43 county-level laboratories in Qinghai Province were organized to participate in the national water fluoride external quality control assessment, and the assessment results were evaluated by Z-ratio scoring method.Results:The response rate of county-level laboratories was 86.05% (37/43) in 2019, and 100.00% in other years. The qualified rate of county-level laboratories was 100.00% from 2016 to 2018; in 2019 and 2020, the assessment was fully covered, and the qualified rate was 81.40% (35/43) and 95.35% (41/43), respectively. Compared with 2019, the response rate and qualified rate in 2020 increased significantly, and the differences were statistically significant (χ 2 = 6.450, 4.074, P < 0.05). In the past 5 years, two assessment samples│Z│ < 1 in each laboratory were the most common, but with two assessment samples 2≤│Z│ < 3 in some qualified laboratories. Conclusion:The consistency of water fluoride determination in Qinghai Province is not very ideal, and the detection ability of county-level laboratories still needs to be strengthened.