Objective To analyze the radiographic characteristics of right hand X-ray film of adult patients with Kaschin-Beck disease (KBD) in Qinghai Province, to understand the most affected locations in adult KBD. Methods According to the criteria of KBD diagnose (WS/T 207-2010), 111 cases of patients were taken X-ray films of right hands. Joint space narrow, joint deformity, subchondral sclerosis, osteophyte, coarse and irregularity of joint, marginal retraction sign and capsule changes were chosen as the descriptive indexes, and these indexes were analyzed with SPSS 17.0. Results A total of 111 cases adult patients with KBD were examined right hand by X-ray, abnormality on X-ray film were 103 cases, the abnormal rate was 92.79%. The most affected fingers were Ⅱ- Ⅳphalanx bones, Ⅱphalanx bones accounted for about 92.23% (95/103), Ⅲ phalanx bones accounted for about 99.03% (102/103), and Ⅳ phalanx bones accounted for about 99.03% (102/103). There was significant difference of the abnormality between th e proximal phalanx and the middle phalanx among the Ⅱ - Ⅳ phalanx bones(χ2=79.33, P<0.05). Abnormal numbers of joint deformity, marginal retraction sign, coarse and irregularity of joint, osteophyte, capsule changes and joint space narrow in the proximal phalanx were 212, 7, 134, 47, 15 and 115 in Ⅱ - Ⅳ proximal phalanx, respectively; while the abnormal numbers of joint deformity, marginal retraction sign, coarse and irregularity of joint, osteophyte, capsule changes and joint space narrow in the middle phalanx were 77, 37, 137, 26, 19 and 126 in Ⅱ - Ⅳmiddle phalanx, respectively. Conclusion The Ⅱ - Ⅳ phalanx bones of right hand are the most affected locations in adult KBD.

OBJECTIVE: To observe the changes of plasminogen activator inhibitor-1 and D-dimer during continuous blood purification (CBP) and related factors. METHODS: Sixteen patients who were diagnosed with multiple organ dysfunction syndrome (MODS) were randomly divided into 2 groups: 8 patients received standard continuous blood purification with heparin anticoagulation, and the other 8 received CBP without anticoagulation. Ten normal blood samples were collected from healthy volunteers as controls. All patients underwent CBP for 8 h. Blood was taken from those patients at 0, 15, 60, 120 and 480 min during the CBP. Plasma plasminogen activator inhibitor-1, D-dimer and serum TNF-α and IL-1β were measured by ELISA. RESULTS: Plasma levels of PAI-1 and D-dimer were increased significantly compared with those in the control group (P<0.05). Plasma level of PAI-1 was reduced (P<0.05) and D-dimer was increased (P<0.05) after the CBP. The level of plasma PAI-1 in the heparin group was significant reduced compared with the group of CBP without anticoagulation (P<0.05). There was negative correlation between the level of PAI-1 and the dosage of heparin used during a CBP session in the heparin group (r=-0.746, P<0.001). CONCLUSION The level of PAI-1 and D-dimer is higher in patients with MODS than that in the normal controls. After the CBP treatment, there is significant decrease in PAI-1 and increase in D-dimer in both groups. Heparin used during CBP can reduce PAI-1 which intensifies its function of anticoagulation.
Anticoagulants ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; analysis ; Heparin ; therapeutic use ; Humans ; Interleukin-1beta ; blood ; Plasminogen Activator Inhibitor 1 ; blood ; Renal Dialysis ; Tumor Necrosis Factor-alpha ; blood

Objective To study the effects of pressure controlled ventilation(PCV)and volume controlled ventilation(VCV)on diaphragm function and the incidence of postoperative pulmonary complica-tions(PPCs)in patients undergoing laparoscopic surgery.Methods Sixty-six patients underwent laparo-scopic gynecological surgery under general anesthesia,aged 18-64 years,BMI 18-30 kg/m2,ASA physical status Ⅰ or Ⅱ were recruited.The patients were randomly divided into two groups:PCV group(group P)and VCV group(group V),33 cases in each group.All the patients were ventilated in VCV mode after induction.Group P was switched to PCV after pneumoperitoneum and group V maintained VCV until the end of operation after pneumoperitoneum.The diaphragm ultrasonic evaluation indexes including di-aphragmatic excursion(DE),diaphragm contraction velocity(DCV),and diaphragmatic rapid shallow breathing index(D-RSBI)were recorded before anesthesia induction,immediately after extubation,and 30 minutes after extubation.The mechanical ventilation time,artificial pneumoperitoneum time,the time from the end of artificial pneumoperitoneum to extubation,the cumulative dosage of cisatracuriumbesylate,and the patient's observer's assessment alert/sedation(OAA/S)immediately after extubation,the incidence of diaphragm dysfunction immediately after extubation and 30 minutes after extubation,and the cumulative in-cidence of PPCs in 1-3 days after operation.Results Compared with group V,DE in group P was in-creased significantly immediately after extubation(P＜0.05),but there was no significant difference in DE of 30 minutes between the two groups after extubation.Compared with group V,DCV in group P was in-creased significantly immediately after extubation and 30 minutes after extubation(P＜0.05),the inci-dence of PPCs in group P was significantly lower on the 1st day after operation(P＜0.05).There were no significant differences in D-RSBI,time of mechanical ventilation,time of artificial pneumoperitoneum,time from the end of pneumoperitoneum to extubation,cumulative dosage of atracurium besylate,OAA/S score immediately after extubation,and the incidence of diaphragm dysfunction immediately after extubation and 30 minutes after extubation,and the cumulative incidence of PPCs on the 2nd and 3rd day after operation.Conclusion In lower abdominal endoscopic gynecological surgery,compared with volume-controlled venti-lation mode,pressure-controlled ventilation mode dose not reduce the incidence of postoperative diaphragm dysfunction,but dose alleviate the weakening of diaphragm inspiratory force and reduce the incidence of pul-monary complications on the first day after operation.

Objective:To understand the situation of drinking water-borne endemic fluorosis (short for drinking water fluorosis) in Huangzhong, Ping'an and Ledu districts of Qinghai Province and operation status of water improvement projects, and to evaluate the effect of control measures.Methods:In historical drinking water fluorosis villages of Huangzhong, Ping'an and Ledu districts of Qinghai Province in 2018, the status of water improvement, the operation of water improvement projects and the content of water fluoride were investigated; the dental fluorosis of 8-12 years old children in all investigated villages was checked; 3 villages from 3 districts were selected, X-ray examination for skeletal fluorosis and urinary fluoride detection of adults over 25 years old of age were performed.Results:Eight-two historical drinking water fluorosis villages in 3 districts were investigated and all the villages had water improvement projects, in addition, the water improvement projects were operating normally, and the water fluoride content ranged from 0.10 to 0.37 mg/L, which were in line with the drinking water fluoride content standard (< 1.2 mg/L). Totally 2 503 children aged 8-12 years old were examined, the detection rate of dental fluorosis was 6.03% (151/2 503), the index of dental fluorosis was 0.13, the epidemic intensity was negative; the detection rates of dental fluorosis in 3 districts were 6.24% (129/2 068), 4.05% (7/173), 5.73% (15/262), respectively, reaching the control standard (< 30%), and the detection rate of dental fluorosis in 81 villages was less than 30%; 198 adults over 25 years old were examined, the detection rate of skeletal fluorosis was 5.05% (10/198), the geometric mean of urinary fluoride was 0.81 mg/L, and skeletal fluorosis cases were mainly in the age group over 40 years old, all of them were mild cases.Conclusions:The fluoride content in drinking water and the detection rate of children's dental fluorosis are all up to the control standard, and the condition of skeletal fluorosis is significantly reduced after the implementation of water improvement measures in Huangzhong, Ping'an and Ledu districts. It can be seen that the prevention and control of drinking water fluorosis in Huangzhong, Ping'an and Ledu districts have achieved good results, but later management and condition monitoring of water improvement projects should be strengthened to prevent the disease from rebounding.

Objective:To investigate the contents of soil metal elements in the plague high incidence area and resting area in the plague natural foci of Qingnan region in Qinghai Province, and to compare and analyze the differences of the contents of soil metal elements between the two areas.Methods:A total of 69 soil samples were collected from 11 sites in the plague high incidence area (Yushu Prefecture) and resting area (Guoluo Prefecture) in Qingnan region, including 39 samples from high incidence area and 30 samples from resting area. The contents of 12 soil metal elements, such as iron (Fe), magnesium (Mg), calcium (Ca), barium (Ba), cobalt (Co), copper (Cu), manganese (Mn), lead (Pb), rubidium (Rb), titanium (Ti), vanadium (V) and zinc (Zn), were measured by X-ray fluorescence spectrometry and SPSS 20.0 software was used for Spearman correlation and other statistical analysis.Results:The contents of Fe and Ca in the soil of the plague natural foci of Qingnan region were relatively high. The contents of 12 soil metal elements in the high incidence area were significantly higher than those in the resting area ( P < 0.01 or < 0.05). There were statistical significant differences in the contents of Mg, Ca, Ba, Mn, Pb and Zn in the soil among the sample plots of the plague high incidence area ( P < 0.01 or < 0.05). There were statistical significant differences in the contents of Fe, Mg, Ca, Rb and Ti in the soil among the sample plots of the plague resting area ( P < 0.05 or < 0.01). Except for Mg and Ca, Fe was positively correlated with other metal elements in the high incidence area and resting area ( P < 0.01 or < 0.05), and Fe was strongly positively correlated with Rb, Ti and V ( r = 0.780 - 0.838). Mg was only positively correlated with Ca in the high incidence area and resting area ( P < 0.01). Ca was negatively correlated with Mn, Pb, Rb, Ti, V and Zn in the high incidence area, and with Rb and Zn in the resting area ( P < 0.01 or < 0.05). The rest of the metal elements were mostly positively correlated in the high incidence area and resting area. Conclusions:The contents of Fe and Ca are rich in the plague natural foci of Qingnan region in Qinghai Province. There are correlations among most soil metal elements in the high incidence area and resting area. The correlation between the contents of soil metal elements in the plague natural foci and the epidemic of plague is worth further exploring.

Objective:To investigate the molecular mechanism of excessive iodine induced experimental autoimmune thyroiditis (EAT) in mice.Methods:Sixty female non-obese diabetic (NOD) mice were selected and divided into 5 groups according to body weight [(25 ± 3) g] via the random number table method, with 12 mice in each group: control group (group A), 10-fold high iodine group (group B), 100-fold high iodine group (group C), 1 000-fold high iodine group (group D) and 1 000-fold high iodine combined with polyinosinic acid-polycytidylic acid [Poly (I:C)] group (group E). The experiment period was 16 weeks. Mice in each group drank purified water with sodium iodine (NaI) content of 0.000, 0.005, 0.050, 0.500 and 0.500 mg/L, respectively; mice in group E were intraperitoneally injected with Poly (I:C) at week 7 and week 15, respectively. At the end of the 16th week, mice were dissected and blood samples and thyroid tissue were taken. The levels of serum thyroid function indexes [thyroid stimulating hormone (TSH), free triiodothyronine (FT 3), free thyroxine (FT 4), and thyroid peroxidase antibody (TPOAb)] were detected by enzyme-linked immunosorbent assay (ELISA); the pathological changes of thyroid tissue were observed after hematoxylin-eosin (HE) staining; differentially expressed genes in thyroid tissue were detected by RNA-sequencing (RNA-seq), and analyzed by KEGG pathway; mRNA and protein levels of p38, intercellular adhesion molecule-1 (ICAM-1) and chemokine 10 (CXCL10) in thyroid tissue were detected by real-time fluorescence quantitative PCR (qPCR) and Western blotting, respectively. Results:There were statistically significant differences in serum levels of TSH (ng/ml: 6.53 ± 0.86, 6.61 ± 0.82, 7.68 ± 0.55, 7.93 ± 0.60, 8.73 ± 1.60), FT 3 (pg/ml: 59.35 ± 10.16, 53.73 ± 10.96, 46.19 ± 8.03, 41.01 ± 8.67, 34.21 ± 11.75), FT 4 (pg/ml: 136.74 ± 10.06, 124.33 ± 14.34, 101.80 ± 6.78, 91.37 ± 6.75, 73.29 ± 17.31), and TPOAb (U/ml: 130.81 ± 24.53, 145.47 ± 28.89, 166.52 ± 41.59, 199.78 ± 42.19, 201.99 ± 44.03) among the 5 groups of mice ( F = 4.77, 4.96, 23.12, 3.68, P < 0.05). Compared with group A, the serum TSH levels of mice in groups C, D and E were higher, the levels of FT 3 and FT 4 in groups B, C, D and E were lower, and the levels of TPOAb in groups D and E were higher, and the differences were statistically significant ( P < 0.05). HE staining showed that the thyroid follicle lesion in groups D and E was serious, and the EAT phenotype appeared in both groups. The differentially expressed genes were analyzed by KEGG pathway. Compared with group A, 8 metabolic pathways related to thyroid autoimmunity and inflammation were found in groups B, C, D and E. Further analysis found that 3 genes appeared in multiple pathways, namely p38, ICAM-1 and CXCL10. There were significant differences in the mRNA levels of p38, ICAM-1 and CXCL10 in thyroid tissue of the 5 groups of mice ( F = 14.77, 12.76, 16.39, P < 0.05); compared with group A, the mRNA levels of p38 in groups B, C, D and E were higher, and the mRNA levels of ICAM-1 and CXCL10 in groups C, D and E were higher ( P < 0.05). There were significant differences in the protein levels of p38, ICAM-1 and CXCL10 in thyroid tissue of the 5 groups of mice ( F = 7.97, 73.86, 18.02, P < 0.05); compared with group A, the protein levels of ICAM-1 and CXCL10 in groups B, C, D and E were higher ( P < 0.05). Conclusion:Excessive iodine promotes the occurrence and development of EAT in mice by up-regulating the expressions of p38 and ICAM-1 genes that are closely related to thyroid autoimmune and inflammatory responses.

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 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 investigate the effect and mechanism of Feixin Decoction(Astragali Radix,Pericae Semen,Carthami Flos,Descurainiae Semen Lepidii Semen,Paeoniae Radix Rubra,etc.)on monocrotaline-induced pulmonary arterial hypertension(PAH)rats based on peroxisome proliferator-activated receptor-γ/nuclear factor-κB(PPAR-γ/NF-κB)signaling pathway.Methods Forty-eight male SD rats were randomly divided into normal group,model group,Sildenafil group(0.025 g·kg-1)and low-,medium-and high-dose of Feixin Decoction groups(11.7,23.4,46.8 g·kg-1).PAH rat model was established by single intraperitoneal injection of monocrotaline solution(60 mg·kg-1).After 1 hour of modeling,the rats were given intragastric administration once a day for 28 days.Hemodynamic and echocardiographic parameters including right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),right ventricular hypertrophy index(RVHI),pulmonary artery acceleration time(PAAT),pulmonary artery ejection time(PET),tricuspid annular plane systolic excursion(TAPSE),right ventricular internal diameter(RVIDd)and right ventricular anterior wall thickness(RVAWT)were measured in each group.The pathological changes of pulmonary arterioles were observed by HE staining.The expression level of α-smooth muscle actin(α-SMA)in rat pulmonary artery was detected by immunofluorescence.The levels of plasma interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)were detected by ELISA.The expression levels of PPAR-γ/NF-κB signaling pathway-related proteins were detected by immunohistochemistry and Western Blot.Results Compared with the normal group,the RVSP,mPAP,RVHI,RVIDd and RVAWT of the model group were significantly increased(P＜0.01).PAAT,PAAT/PET and TAPSE were significantly decreased(P＜0.01).The wall of pulmonary arterioles was significantly thickened,and the percentage of wall thickness of pulmonary arterioles to vascular diameter and the percentage of vascular wall area to total cross-sectional area of pulmonary arterioles were significantly increased(P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly increased(P＜0.01).The levels of plasma IL-1β,IL-6 and TNF-α were significantly increased(P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly decreased(P＜0.01),and the positive expression rate of NF-κB protein was significantly increased(P＜0.01).The protein expressions of PPAR-γ and IκB-α in lung tissue were significantly down-regulated(P＜0.01).The protein expression ratio of p-NF-κB/NF-κB was significantly increased(P＜0.01).Compared with the model group,RVSP,mPAP,RVHI,RVIDd and RVAWT in each administration group were significantly decreased(P＜0.05,P＜0.01),while PAAT,PAAT/PET and TAPSE were significantly increased(P＜0.05,P＜0.01).The thickness of the vascular wall was significantly reduced,and the percentage of the wall thickness of the pulmonary arterioles to the diameter of the blood vessels and the percentage of the vascular wall area to the total cross-sectional area of the small arteries were significantly reduced(P＜0.05,P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly decreased(P＜0.05,P＜0.01).The plasma levels of IL-1β,IL-6 and TNF-α were significantly decreased(P＜0.05,P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly increased(P＜0.05,P＜0.01),and the positive expression rate of NF-κB protein was significantly decreased(P＜0.05,P＜0.01).The protein expression of PPAR-γ in lung tissue was significantly up-regulated(P＜0.05,P＜0.01),and the protein expression ratio of p-NF-κB/NF-κB was significantly decreased(P＜0.01).The protein expression of IκB-α in the lung tissue of rats in the high-dose group of Feixin Decoction was significantly up-regulated(P＜0.01).Conclusion Feixin Decoction can improve pulmonary artery pressure,right ventricular dysfunction and pulmonary vascular remodeling in PAH rats induced by monocrotaline.The mechanism may be related to the regulation of PPAR-γ/NF-κB signaling pathway to inhibit inflammatory response.

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.