Objective:To clone and express the full length cDNA of human CD38. Methods:The full length cDNA of the human CD38 antigen was amplified from total RNA of Daudi cell by RT-PCR, and it was inserted into pGEM-T. The validity on the sequences was confirmed by automatic DNA sequencing. Inserting the valid CD38 gene into pcDNA3.1(+) plasmid to obtain recombinant mammalian expression vector pcDNA3.1(+)/CD38Z; Using lipofectin gene transfer technique system, recombinant expression vector containing CD38 gene was transfected into COS7 cells. The expression of CD38 molecules on the surface of COS7 cells was detected by FACS and immunohistochemical technique. Results:DNA sequencing showed that the cloned full length cDNA sequence was identical with reported. The result of FACS and immunohistochemical technique indicated that CD38 molecules were expressed on the surface of COS7 cells. Conclusion:The full length cDNA of human CD38 is obtained, recombinant mammalian expression vector pcDNA3.1(+)/CD38Z is successfully constructed, and the CD38 molecules is expressed on the surface of COS7 cells,this may facilitate studies on the biochemistry and function of CD38 antigen.

Objective To investigate the distribution of gut microbiota in obese patients with or without acanthosis nigricans .Methods Totally 131 obese patients and 25 healthy participants were divided into three groups:the obesity with acanthosis nigricans (AN) group (n=59), the simple obesity (OB) group (n=79), and the control (CON) group (n=25).The fresh stool samples were collected , and the clinical and biochemistry markers were measured .Pyrosequencing technology was performed based on the 16s rRNA of fecal samples to identify and analyze the distribution pattern of gut microbiota in each group .Results The AN group had signifi-cantly higher body mass index [ (37.45 ±5.12) kg/m2 vs.(33.34 ±2.54) kg/m2 vs. (20.35 ±1.68) kg/m2, P=0.045, P<0.001], insulin [32.77 (25.18) mU/L vs.20.73 (9.30) mU/L vs.8.70 (6.18) mU/L, P<0.001, P<0.001], insulin resistance [7.78 (6.87) vs.4.71 (2.88) vs.1.81 (1.40), P<0.001, P<0.001], and interleukin (IL) -6 [ (3.64 ±2.23) ng/L vs.(2.71 ±0.78) ng/L vs.(2.17 ±0.86) ng/L, P=0.040, P=0.009] levels than OB and CON groups compared with OB and CON groups , AN group had sig-nificantly decreased diversity of bacterial flora ( P=0.015 , P=0.001 ) , while no significant difference was observed in the abundance of bacterial flora .At the phylum level , the composition of flora among these three groups was similar, mainly including bacteroidetes , firmicutes, proteobacteria, and actinomycetes.Although the proportions of main bacteria flora were different , the difference was not statistically significant .At the genus level, the bacteria flora in AN and OB groups were primarily composed of Bacteroides, Megamonas, Faecalibac-terium and Escherichia-Shigella.In addition, compared to OB and CON groups , AN group had significantly lower proportion of Ruminococcus ( P=0.023 , P=0.043 , respectively ) and higher proportion of Veillonella (P=0.048, P=0.043, respectively).Furthermore, the proportion of Weissella was higher in AN and OB groups than in CON group ( P=0.045 , P=0.025 ) .Conclusion Obese patients with AN have more severe in-sulin resistance and inflammation status than those with simple obesity , and the distribution feature of gut micro-biota also differ between these two patient populations .

Objective To explore the mechanism by which the anti-human P185erbB2 scFv-Fc-IL-2(HFI)modulates tumor surface molecules and activates immune effector cells in vitro. MethodsMTT assay was used to test the proliferation and the LAK-like cytotoxicity. Flow cytometry assay was used to test the expression of ICAM-1, Fas and erbB2 receptors in tumor cells and the expression levels of CD molecules FasL and LFA-1 in human PBMC. Antibody-dependent cell-mediated cytotoxicity(ADCC)mediated by HFI against SKOV3, MCF-7 and SGC-7901 tumor cells was explored hv LDH release assay. Results The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI were upregulated, from 24.85％ and 0.53％ to 85.36％ and 59.19％ respectively, while the expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI were downregulated, from 98.48％, 42.60％ and 36.66％ to 94.01％,30.95％ and 12.36％ respectively. HFI could significantly enhance the proliferation activity of human PBMC, and CD3+ CD8+ T cells and CD3- CD16+ CD56+ NK cells were elevated, from 24.37％ and 6.90％ to 38.80％ and 13.45％ respectively. The expression levels of CD25, LFA-1 and FasL were significantly enhanced from 3.99％, 86.52％ and 5.02％ to 12.96％, 99.06％ and 16.19％. The LAK-like cytotoxicity of human PBMC treated with HFI against SKOV3, MCF-7,SGC-7901 tumor cells was significantly improved:HFI was effective in mediating ADCC against SKOV3,MCF-7 and SGC-7901 tumor cells which expressed high,medium and low levels of erbB2,respectively,and HFI-induced ADCC was correlated with the degrees of erbB2 expression on the tumor cells. Conclusion The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI are significantly upregulated. The expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI are downregulated. HFI can significantly enhance the proliferation activity of human PBMC. The LAK-like eytotoxicity of human PBMC treated with HFI against tumor cells is significantly enhanced. HFI iS effective in mediating ADCC and the activity of HFI-induced ADCC is correlated with the degrees of erbB2 expression on the tumor cells.

Objective To assess the variation of sexual hormone and mechanisms of low testosterone in young male obesity with acanthosis nigricans. Methods Retrospective analysis was performed in 125 male obesity patients [ body mass index( BMI)≥28 kg/m2 ] . According to their clinical characteristics, they were divided into two groups including obesity without acanthosis nigricans(OB group, n=62) and obesity with acanthosis nigricans(AN group, n=63). 60 normal weight men were also recruited as a control group. Body fat and body weight were measured. Blood insulin, lipid profile, sex hormones levels, and inflammation factors were measured. Parameters of each group were compared and the correlations between total testosterone level and other index were analyzed. Results All the male obesities have the significant lower total testosterone levels than those of control group(P>0. 05), and those in AN group were lower than those in OB group(P>0. 05). The BMI and body fat in OB group and AN group were both significantly higher than those in control group(P>0. 05). The fasting insulin levels in all obese men were significantly higher than those in control group(P>0. 05), highest in AN group. Triglycerides(TG) in both OB and AN group were higher than those in controls, and not significant between later 2 groups. But high-density lipoprotein-cholesterol ( HDL-C) in the two groups were significantly lower than control, which in AN group were significantly lower than OB group. Total testosterone levels in AN group were negatively correlated with weight, waist circumference, hip circumference, fasting insulin, and homeostasis model assessment for insulin resistance ( HOMA-IR ) , and also negatively correlated with inflammation factors including C-reactive protein ( CRP ) , erythrocyte sedimentation rate ( ESR) , tumor necrosis factor-α( TNF-α) , and uric acid. However, total testosterone levels in AN group were not correlated with lipid metabolism index. Conclusion Young male obesity with acanthosis are associated with secondary hypogonadism. Hyperinsulinemia, insulin resistance, and inflammatory factors are risk factors for the occurrence of this secondary male hypogonadism.

Objective:To verify the revised method of cerium sulfate catalytic spectrophotometry for iodide index of "Standard Examination Methods for Drinking Water-Nonmetal Parameters" (GB/T 5750.5-2006).Methods:From July to September 2019, the Laboratory of Department for Endemic Disease Prevention and Control of Qinghai Institute for Disease Prevention and Control verified the revised method (determination of iodide in drinking water by cerium sulfate catalytic spectrophotometry) of cerium sulfate catalytic spectrophotometry (hereinafter referred to as original method) in "Standard Examination Methods for Drinking Water-Nonmetal Parameters" (GB/T 5750.5-2006). The revised method was verified according to the requirements of "Standard Examination Methods for Drinking Water-Water Analysis Quality Control" (GB/T 5750.3-2006), including standard curve, detection limit, precision, accuracy and actual sample determination.Results:The linear range of the revised method was 0 - 20.0 μg/L, the correlation coefficient was - 0.999 4 - 0.999 8, and the detection limit was 0.231 μg/L. The relative standard deviation ( RSD) of low, medium and high iodine water samples of 6 times detection ranged from 1.4% to 9.6%, and the recoveries of low and medium water samples ranged from 89.0% to 108.0%. The detection results of national first-class reference materials for iodine composition analysis in water were within the range of standard value ± uncertainty. There was no significant difference in the test of results of 12 tap water samples between the revised method and the original standard method ( t = - 0.075, P > 0.05). Conclusion:The revised method has a good linear relationship of standard curve, high precision and accuracy, and good reproducibility, is simple and easy to operate, and is suitable for promotion and application.

Objective:To investigate the iodine nutritional status of children in Qingdao City and the effects of prevention and treatment measures on iodine deficiency disorders (IDD), and to provide a scientific basis for guiding residents to scientifically supplement iodine, taking timely targeted prevention and control measures, and scientifically adjusting intervention strategies.Methods:According to "National Iodine Deficiency Disorders Surveillance Program (2016 edition)" and "Iodine Deficiency Disorders Surveillance Program of Shandong Province", from 2018 to 2020, using the cluster sampling method, children aged 8-10 years in Qingdao City were chosen to test their household salt iodine content and random urinary iodine content, and to examine their thyroid volume by B-ultrasonography, and the correlation between thyroid volume and physical development indexes was analyzed.Results:From 2018 to 2020, a total of 6 057 children were monitored, including 3 068 boys and 2 989 girls. The median of salt iodine and iodized salt iodine of children was 23.50 and 24.10 mg/kg. The qualified rate of iodized salt was 89.95% (4 832/5 372), the coverage rate of iodized salt was 88.69% (5 372/6 057), and the consumption rate of qualified iodized salt was 79.78% (4 832/6 057). There were significant differences in the qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt between different years (χ 2 = 135.26, 314.71, 342.87, P < 0.001). A total of 6 057 urine samples were collected from children, and the median of urinary iodine was 193.92 μg/L, of which 16.2% (979/6 057) were < 100 μg/L, and 22.5% (1 361/6 057) were ≥300 μg/L. There were statistically significant differences in the medians of urinary iodine between different years, gender and whether eating iodized salt ( H/Z = 37.25,-3.89,-5.69, P < 0.001), the median of urinary iodine in boys was higher than that of girls, and the median of urinary iodine in eating iodized salt group was higher than that of eating non-iodized salt group. There was no significant difference in the median of urinary iodine between different age ( H = 4.33, P = 0.119). The rate of goiter in children was 3.45% (71/2 057), and the difference between different years was statistically significant (χ 2 = 42.68, P < 0.001). The incidence of goiter in 2020 [7.31% (45/616)] was significantly higher than that in 2018 and 2019 [2.81% (18/641), 1.00% (8/800), P < 0.001]. Thyroid volume of children was positively correlated with height and weight ( r = 0.20, 0.22, P < 0.001). Conclusions:The iodine nutritional level of children aged 8-10 years in Qingdao City is appropriate. However, the incidence of goiter in children in some years is relatively high. The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt are all lower than the national standard for elimination of IDD, which should be paid attention to.

Objective:To study the epidemic status of drinking tea type endemic fluorosis in Qinghai Province.Methods:In 2019, in counties (cities, districts, referred to as counties) that had the habit of drinking brick tea in 8 cities (prefectures) of Qinghai Province, epidemiological investigation of drinking tea type endemic fluorosis was carried out in villages. Ten households were randomly selected from each village, to investigate the demographic data of each household and the drinking situation of brick tea, residents' drinking water and brick tea samples were collected to determine the fluorine content, and calculate the daily per capita tea fluorine intake. At the same time, skeletal fluorosis was examined in all adults over 25 years old, and dental fluorosis was examined in all children aged 8 to 12 years old in survey sites. The content of fluorine in tea and water was detected by ion selective electrode method; the diagnosis of skeletal fluorosis was based on "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008), the diagnosis of dental fluorosis was based on "Diagnosis of Dental Fluorosis" (WS/T 208-2011).Results:The mean (range) of fluorine of the 3 602 water samples was 0.31 (0.20 - 1.00) mg/L. The geometric mean (range) of fluorine of the 31 067 brick tea samples was 646 (40 - 2 295) mg/kg, the fluorine content of the brick tea ≤300 mg/kg accounted for 7.80% (2 422/31 067) of the total samples. The proportion of drinking Fu brick-tea was 89.97% (27 952/31 067); and the daily per capita tea fluorine intake was 1.93 mg, the daily per capita tea fluorine intake in Guoluo, Yushu and Hainan prefectures were higher than the health standard (3.50 mg). The detection rate of skeletal fluorosis in adults was 0.16% (2 357/1 484 907), Yushu Prefecture was the highest [29.23% (592/2 025)], followed by Guoluo Prefecture, which was 8.21% (771/9 393). The detection rate of dental fluorosis in children was 4.79% (8 076/168 623), Yushu Prefecture was the highest [32.61% (1 562/4 790)].Conclusion:Drinking tea type endemic fluorosis is prevalent in Qinghai Province, with obvious regional characteristics, covering a large population. The disease is relatively popular in Yushu Prefecture and Guoluo Prefecture.

Objective:To assess the operation and usage conditions of all water improvement projects in Qingdao, master the water fluoride content and the prevalence of dental fluorosis in children in drinking-water-borne endemic fluorosis villages, evaluate the prevention and control effect of water improvement project.Methods:From April to October in 2020, all drinking-water-borne endemic fluorosis villages in 7 cities (districts) of Qingdao were selected to investigate the operation and usage conditions of water improvement project, water fluoride and prevalence of dental fluorosis of 8 to 12 years old children. The prevention and control effect of water improvement project on drinking-water-borne endemic fluorosis in Qingdao was evaluated according to the control contents and criteria of drinking-water-borne endemic fluorosis.Results:There were 1 146 drinking-water-borne endemic fluorosis villages, the water improved rate of villages was 100.00% (1 146/1 146), the rate of normal operation of water improvement project was 99.91% (1 145/1 146). There were 1 130 villages with qualified water fluoride and the qualified rate was 98.60% (1 130/1 146). A total of 23 473 children aged 8 - 12 years old were examined, and 631 children were detected with dental fluorosis. The detection rate of dental fluorosis was 2.69% (631/23 473), the dental fluorosis index was 0.048, and the prevalence was negative. The detection rate of dental fluorosis of children in villages with excessive water fluoride (7.03%, 40/569) was higher than that in villages with qualified water fluoride (2.58%, 591/22 904, χ 2 = 41.235, P < 0.01). Conclusion:The operation and usage conditions of water improvement projects in 7 drinking- water-borne endemic fluorosis cities (districts) in Qingdao are good, and the detection rate of dental fluorosis in children is low, which has reached the national control standard.

Objective:To evaluate the effect of prevention and control measures to drinking-water-borne endemic fluorosis in Qingdao, and to provide scientific basis for formulating prevention and control strategies.Methods:From 2019 to 2021, a full coverage surveillance was carried out in 1 146 villages in 7 drinking-water-borne endemic fluorosis areas in Qingdao, including the operation status of the water improvement and fluoride reduction project, the water fluoride content in the villages, and the prevalence of dental fluorosis among children aged 8-12 years.Results:From 2019 to 2021, all the 1 146 affected villages in Qingdao had improved their water supply, and the rates of water fluoride exceeding the standard were 7.16% (82/1 146), 1.40% (16/1 146) and 3.84% (44/1 146), respectively, and the differences between years were statistically significant (χ 2 = 48.36, P < 0.001). The detection rates of dental fluorosis in children aged 8-12 years were 3.11% (803/25 856), 2.68% (629/23 460) and 3.00% (655/21 846), respectively, and the differences between years were statistically significant (χ 2 = 8.26, P = 0.016). The detection rate of dental fluorosis among children in villages with qualified water fluoride (2.85%, 1 986/69 565) was lower than that in villages with excessive water fluoride (6.32%, 101/1 597), with a statistically significant difference (χ 2 = 67.74, P < 0.001). Conclusions:From 2019 to 2021, the effect of prevention and control measures to drinking-water-borne endemic fluorosis in Qingdao is significant and the detection rate of dental fluorosis in children is low, the detection rate of dental fluorosis in villages with qualified water fluoride is significantly lower than that in villages with excessive water fluoride. In the future, we should continue to strengthen water fluoride monitoring and engineering maintenance to ensure that the water fluoride content continues to be qualified and effectively reduce the harm of fluorosis.

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.