[A clinical challenge of evaluation and management in children with genetic obesity].

Obesity is a growing global public health problem, while 40%-70% of obesity is determined by genetic factors. This article focuses on the classification, disease characteristics, diagnosis and progressive treatment of childhood genetic obesity. The prevention and control of childhood genetic obesity requires early detection of obese individuals and early screening of obesity causes. At the same time, clinicians are advised to propose individualized therapy and intervention measures based on multi-disciplinary opinions to improve the health of genetic obese children.

[The construction of comprehensive stroke prevention and control system requires attention to the precise policies of young stroke].

Youth is the core force of social and economic development, once the occurrence of youth stroke will place a heavy burden on society and family. However, the prevention and control of stroke in China is mainly aimed at middle-aged and elderly patients, the part of young stroke is relatively easy to be ignored. This article focuses on the characteristics, research progress, prevention and control status of young stroke, pointing out the importance of centering on the prevention and treatment of young stroke. At the same time, it hopes that the industry can concentrate on the prevention and treatment of young stroke, making precise policies in the future, and developing secondary prevention guidelines for the causes or risk factors of young stroke, so as to improve comprehensive stroke prevention and control system. On this basis, the health level of the whole population will be improved, and the life expectancy of residents will be extended, thus promoting the realization of the strategic goal of "Healthy China 2030".

[The preliminary report of a registration clinical trial of proton and heavy ion irradiation].

Objective: To verify the safety and efficacy of IONTRIS particle therapy system (IONTRIS) in clinical implementation. Methods: Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial: 31 males and 4 females with a median age of 69 yrs (range 39-80). Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non-metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results: Twenty-two patients received carbon ion and 13 had proton irradiation. With a median follow-up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression-free survival rate was 93.8% (15/16). Among the 19 patients with prostate cancer, biological-recurrence free survival was 100%. Particle therapy was well tolerated in all 35 patients. Twenty-five patients (71.4%) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow-up. Six (17.1%) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions: IONTRIS is safe and effective for clinical use. However, long term follow-up is needed to observe the late toxicity and long term result.

[Effect of calcium-independent phospholipase A(2) inhibitor in reducing hepatocyte lipoapoptosis and improving insulin resistance].

Objective: To investigate the effect of calcium-independent phospholipase A(2) (iPLA(2)) inhibitor in reducing hepatocyte lipoapoptosis and improving insulin resistance. Methods: A total of 28 male Sprague-Dawley rats were randomly divided into the following three groups: 12 rats in group I (normal control group) were given normal diet for 18 weeks; 8 rats in group II (non-alcoholic fatty liver disease model group) were given high-fat diet for 18 weeks; 8 rats in group III (iPLA(2) inhibitor group) were given high-fat diet for 18 weeks and intraperitoneal injection of the iPLA(2) inhibitor bromoenol lactone 150 µg/kg once every other day since week 15 (14 times of injection in total). All the rats were sacrificed at the same time, and body weight and liver weight were measured. Blood lipids, serum enzymes, fasting blood glucose, fasting insulin, free fatty acid, and serum iPLA(2) concentration were measured in each group, and liver pathological changes were evaluated. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to measure the level of hepatocyte apoptosis and the apoptotic index was calculated. Quantitative PCR was used to measure the mRNA expression of iPLA(2). The Student-Newman-Keuls test and the chi-square test were used for comparison of parameters between groups I, II, and III. P < 0.05 was considered statistically significant. Results: Compared with group I, group II had significant increases in triglyceride (0.75±0.05 mmol/L vs 1.20±0.13 mmol/L, P < 0.05), cholesterol (1.50±0.12 mmol/L vs 2.94±0.34 mmol/L, P < 0.05), low-density lipoprotein (0.65±0.06 mmol/L vs 1.30±0.16 mmol/L, P < 0.05), free fatty acid (0.58±0.09 mEq/L vs 0.80±0.20 mEq/L, P < 0.05), fasting blood glucose (4.85±0.22 mmol/L vs 6.94±0.65 mmol/L, P < 0.05), and fasting insulin (0.89±0.52 mmol/L vs 1.29±0.52 mmol/L, P < 0.05), and a significant reduction in the insulin sensitivity index (0.52±0.21 vs 0.27±0.11, P < 0.05); group II also had significant inflammation and fatty degeneration shown by liver pathology, and compared with group I, group II had significant increases in apoptotic cells and apoptotic index (0.58%±0.17% vs 39.69%±4.96%, P < 0.05). Compared with group I, group II had significant increases in serum iPLA(2) concentration (2.92±0.08 ng/ml vs 3.28±0.14 ng/ml, P < 0.05) and the mRNA expression of iPLA(2) in the liver (1.07±0.18 vs 7.68±0.49, P < 0.05). Compared with group II, group III had a lower level of hepatocyte apoptosis, a significant reduction in apoptotic index (39.69%±4.96% vs 24.80%±2.53%, P < 0.05), significant reductions in serum iPLA(2) concentration (3.28±0.14 ng/ml vs 2.64±0.24 ng/ml, P < 0.05) and the mRNA expression of iPLA(2) in the liver (7.68±0.49 vs 2.60±0.36, P < 0.05), significant reductions in fasting insulin (1.29±0.52 mmol/L vs 0.80±0.09 mmol/L, P < 0.05) and fasting blood glucose (6.94±0.65 mmol/L vs 5.18±0.35 mmol/L, P < 0.05), and a significant increase in insulin sensitivity index (0.27±0.11 vs 0.45±0.09, P < 0.05). Conclusion: There is a significant increase in the expression of iPLA(2) in rats with non-alcoholic fatty liver disease, and iPLA2 inhibitor can reduce hepatocyte lipoapoptosis and improve insulin resistance.

[Value of intravoxel incoherent motion diffusion-weighted imaging in differential diagnosis of benign and malignant hepatic lesions and blood perfusion evaluation].

Objective: To investigate the value of intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) in the differential diagnosis and blood perfusion evaluation of benign and malignant hepatic lesions. Methods: A retrospective analysis was performed for 86 patients (96 lesions) with pathologically or clinically confirmed hepatic lesions or hepatic lesions diagnosed based on follow-up results, among whom 48 had malignant lesions (53 lesions) and 38 had benign lesions (43 lesions). The patients underwent conventional magnetic resonance (MR) plain scan, contrast-enhanced scan, and diffusion-weighted imaging (DWI) with different b values (b = 0, 50, 100, 150, 200, 400, 600, 800, 1 000, and 1 200 s/mm2) to determine the parameters of the double exponential model for intravoxel incoherent motion (IVIM): fast diffusion coefficient Dfast, slow diffusion coefficient Dslow, and percentage of fast-diffusion constituent F value. The patients were divided into groups according to the blood supply to lesions on conventional MR plain scan and contrast-enhanced scan, and there were 47 lesions in abundant blood supply group and 49 in poor blood supply group. The data for analysis were Dfast, Dslow, and F values of benign/malignant lesion groups and abundant/poor blood supply groups. The independent samples t-test was used for statistical analysis; the independent samples non-parametric test Mann-Whitney U test was used for the comparison of F value; the receiver operating characteristic (ROC) curve was used to evaluate the value of above parameters in the differentiation of benign and malignant lesions and blood supply evaluation. Results: Compared with the malignant lesion group, the benign lesion group had significantly higher Dslow, and F values (P< 0.001 orP= 0.001) and a higher Dfast value (P= 0.053). Compared with the poor blood supply group, the abundant blood supply group had significantly higher Dfast and F values (P< 0.001 orP= 0.001) and a higher Dslow value (P= 0.185). According to the ROC curve, the cut-off values of Dslow, Dfast, and F values in the diagnosis of benign/malignant hepatic lesions and evaluation of abundant/poor blood supply were 1.18×10-3mm2/s, 27.20×10-3mm2/s, 20.25%, 1.17×10-3mm2/s, 20.30×10-3mm2/s, and 17.80%, respectively, with sensitivities, specificities, accuracy, and areas under the ROC curve of 90.69%/92.45%/91.66%/0.938, 46.51%/73.58%/61.45%/0.589, 74.41%/50.94%/62.50%/0.653, 59.57%/57.14%/58.33%/0.559, 55.32%/63.26%/59.37%/0.618, and 93.61%/89.79%/90.62%/0.961, respectively. Conclusion: The parameter of the double exponential model for IVIM, Dslow value, has a certain value in the differential diagnosis of benign and malignant hepatic lesions, and F value can show blood perfusion in benign and malignant hepatic lesions without the need for contrast-enhanced scan, which provides a reference for the qualitative diagnosis of liver tumor.

Association between obesity and sexual maturation in Chinese children: a muticenter study.

OBJECTIVE: We aimed to evaluate the current status of sexual maturation of Chinese children, to examine the association between obesity and early sexual maturation in boys and compare it with girls and to test the hypothesis that the associations differ by gender. STUDY DESIGN: Cross-sectional study. SUBJECTS: A representative sample involving 9812 boys and 8895 girls aged 6-18 years who participated in the Chinese Children and Adolescent Metabolic Syndrome Epidemiologic Study (July 2009- July 2010) were surveyed. METHODS: All subjects had complete anthropometry and sexual maturation data. SUBJECTS who reached Tanner stage 2 or more (5601 boys and 6538 girls) were divided into tertiles based on the timing of sexual maturation. The subjects in the earliest tertile were included into the early-maturing group, and the middle tertile and the latest tertile into the not early-maturing group. Overweight was defined as a body mass index (BMI) ⩾85th percentile and obesity ⩾95th percentile. Logistic regression analysis was used to test how early maturation affected the risk of overweight. Multiple linear regression was used to examine the association between fatness (BMI Z-score) and sexual maturation. RESULTS: Slightly more boys were obese than girls (P<0.01). The median age for girls of the Tanner stage 2 was 9.69 years, and for boys of Tanner stage 2 was 11.25 years. BMI Z-score were higher (P<0.01) in both early-maturing girls and boys, compared with the non-early maturers, respectively. Early sexual maturation was positively associated with obesity in both girls and boys. With covariates adjusted and using non-early maturing as the reference group, odds ratios for combined overweight were 1.48 for boys and 2.64 for girls, and for obesities were 1.61 for boys and 3.49 for girls, respectively. CONCLUSION: Obesity is positively associated with sexual maturation in both boys and girls, and the association does not differ by gender, but the association is stronger in girls than in boys.

First Report of Anthracnose of Malva sylvestris Caused by Colletotrichum trifolii in China.

Common mallow (Malva sylvestris L.) is a perennial medicinal plant in the Malvaceae family, which is native to Asia, Europe, and North Africa. In July 2012, typical symptoms of anthracnose disease, with a disease incidence of ~70%, were observed on common mallow in the Medicinal Herb Garden of Shenyang Pharmaceutical University, Liaoning, China. The fungus mainly infected the stalks and leaves of M. sylvestris. Pinpoint, brownish lesions initially appeared at the flowering stage and the disease spread within the field. The lesions on stems gradually enlarged and became dark brown, elliptical, and slightly concave. Concurrently, acervuli and mucilaginous conidial masses of the pathogen appeared on lesions under moist conditions. Conidia were hyaline, one-celled, cylindrical with both ends rounded, and measured 10.0 to 12.5 × 2.5 to 4.0 µm (mean 11.3 × 3.3 µm). The fungus was isolated from symptomatic tissues. Small pieces from leaves and stems were surface disinfested with 70% ethanol and 1.5% sodium hypochlorite for 1 min, then rinsed three times with sterile distilled water, and cultured on potato dextrose agar (PDA) at 25°C. The colonies on PDA had initially white aerial mycelia, and later became greenish black with regularly whorled rings. To confirm Koch's postulates, five 3-month-old plants of M. sylvestris were inoculated with a conidial suspension (105 conidia/ml) prepared from PDA cultures incubated for 14 days. Five non-inoculated plants served as controls. The plants were maintained in the greenhouse at 22 to 25°C and about 75% relative humidity under natural daylight. Typical symptoms on inoculated plants were reproduced after ~10 to 14 days, whereas control plants remained asymptomatic. The pathogen was successfully recovered from symptomatic tissues and re-identified, completing Koch's postulates. The internal transcribed spacer (ITS) and large subunit -28S (LSU) region of rDNA was amplified with primers ITS1/ITS4 and NL1/NL4, respectively, and sequenced. Phylogenetic trees (ITS and LSU) that were obtained using MEGE3.1 with the neighbor-joining method showed that both of the isolates fall in the Colletotrichum trifolii clade. The representative sequences (ITS and LSU) were deposited in GenBank (Accession Nos. KJ155692 and KJ920935). The fungus isolated from symptomatic tissues was identified as C. trifolii on the basis of morphological, cultural characteristics, and sequence analysis (2). According to previous references, C. orbiculare and C. malvarum on Malvaceae were respectively described in America and Europe (1,3,4). However, the isolate from M. sylvestris significantly differed from those of C. orbiculare and C. malvarum in cultural characteristics and sequence analysis. In this paper, the results showed that M. sylvestris is a new host of C. trifolii. To our knowledge, this is the first report of mallow anthracnose caused by C. trifolii in China. References: (1) J. A. Bailey et al. Phytopathology 86:1076, 1996. (2) U. Damm et al. Fungal Divers. 61:29, 2013. (3) K. Hyde et al. Fungal Divers. 39:147, 2009. (4) L. Tosi et al. Plant Dis. 88:425, 2004.

Aliskiren attenuates proteinuria in mice with lupus nephritis by a blood pressure-independent mechanism.

This study revealed that low-dose aliskiren treatment could attenuate proteinuria by interrupting the renin-angiotensin system in mice with lupus nephritis, and the beneficial effect was beyond blood pressure control. An in and ex vivo fluorescence imaging (using a non-invasion in vivo imaging system) showed intense labeling of renin in the kidneys of female MRL/lpr mice. In the study, Alzet mini-osmotic pumps were implanted into 6-week-old female MRL/lpr mice. Pumps were filled with either phosphate-buffered saline or a solution of aliskiren dissolved in phosphate-buffered saline (20 mg/kg/day) and replaced at 28-day intervals. Mice were sacrificed at four and eight weeks. To label cells for DNA synthesis, bromodeoxyuridine (BrdU) (50 mg/kg) was injected intraperitoneally an hour prior to sacrifice. The level of renin inhibition was adequate, as aliskiren-treated mice demonstrated higher renal renin mRNA expression than controls (p < 0.05). Although there were no significant differences in the systolic blood pressure (control versus aliskiren-treated: 127.20 ± 4.44 mmHg versus 103.80 ± 7.40 mmHg, p > 0.05) and heart rate (control versus aliskiren-treated: 680.50 ± 11.71 versus 647.80 ± 13.90, p > 0.05) of both groups after eight weeks, there was significant reduction of inflammatory cytokines (transforming growth factor-beta1, regulated on activation normal T cell expressed, monocyte chemoattractant protein-1 and osteopontin, p < 0.05), reduction of innate immunity (toll-like receptor 7, p < 0.05), as well as a reduction of glomerular proliferation and inflammation (BrdU-, CD45-, CD3- and F4/80-positive glomerular cells, p < 0.01) after aliskiren infusion, which might translate into an improvement in proteinuria (control versus aliskiren-treated: 493.7 versus 843.7 mg/g, p < 0.01) or weight gain (control versus aliskiren-treated: 5.65 ± 1.61 versus 8.67 ± 0.97%, p < 0.05).

First Report of Root Rot on Pulsatilla koreana Caused by Fusarium oxysporum in China.

Windflowers (Pulsatilla spp.) are perennial medicinal plants in the family Ranunculaceae with high economic as well as medicinal value in China. It is commonly used as traditional Chinese medicine (1). In May 2012, a root rot disease was observed on windflower (Pulsatilla koreana Nakai) at flowering stages in fields of Liaoning Province, China. The diseased area was estimated to be over 500 ha in the province and the yield was reduced by 30% on average with up to 45% yield losses in some fields. As the disease progressed, brown lesion production extended onto lateral and main roots, and aboveground tissues shriveled and decayed; in severe cases, white mycelium was clearly visible on diseased root tissue. Isolations from symptomatic roots were made on potato dextrose agar (PDA) and single-spore cultures were obtained. Colonies were initially white, but became pale violet with age, and purple pigments were produced in the agar. Microconidia were abundant, unicellular, oval to reniform, and ranged from 5.6 to 13.1 (9.3) × 2.8 to 4.2 (3.2) µm. Macroconidia were sparse, three-septate, slightly curved, and ranged from 21.9 to 39.4 (31.2) × 3.4 to 4.5 (3.9) µm. The isolated fungus was morphologically similar to Fusarium oxysporum (2). Two isolates were selected for molecular identification, and the internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (3) and sequenced. The obtained sequences (GenBank Accession Nos. JX669525 and JX669526) showed 99% homology with the sequences of F. oxysporum in GenBank (GQ121303). For pathogenicity tests, the isolate was cultured on PDA for 10 days at 25°C. Inoculations were performed on 10 healthy P. koreana plants by spraying a conidial suspension (2.0 × 105 microconidia ml-1) on roots previously wounded with a metal needle. Ten non-treated plants used as controls were sprayed with distilled water. The inoculated plants were incubated at 25°C under conditions of 12/12 h (light and dark). After 2 weeks, root rot symptoms were similar to the original symptoms observed under field conditions. No disease was observed on water-inoculated control plants. The same fungus was reisolated from the roots of infected plants, satisfying Koch's postulates. To our knowledge, this is the first report of F. oxysporum on P. koreana in China. The disease was hitherto scarcely reported in any other countries, and may deserve more attention in the future. References: (1) S. C. Bang et al. J. Nat. Prod. 68:268, 2005. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Professional, Ames, IA, 2006. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

[Characteristics of serum bile acids among healthy children in Zhejiang province].

Objective: To characterize the serum bile acid profiles of healthy children in Zhejiang Province. Methods: A cross-sectional study was conducted on 245 healthy children who underwent imaging and laboratory biochemical tests during routine physical examinations at the Children's Hospital of Zhejiang University School of Medicine from January 2020 to July 2022. Overnight fasting venous blood samples were collected, and the concentrations of 18 individual bile acids in the serum were accurately quantitated using tandem mass spectrometry. The concentration difference of bile acid were compared between different genders and to explore the correlation between age and bile acid levels. Used the Mann-Whitney U test for intergroup comparison and Spearman test to correlation analysis. Results: A total of 245 health children with a age of 10 (8, 12) years including 125 boys and 120 girls. There were no significant differences in levels of total bile acids, primary and secondary bile acids, free and conjugated bile acids between the two gender groups (all P>0.05). The serum concentrations of ursodeoxycholic acid and glycoursodeoxycholic acid in girls were significantly higher than those in boys (199.0 (66.9, 276.5) vs. 154.7 (49.3, 205.0) nmol/L, 274.0 (64.8, 308.0) vs. 181.0 (43.8, 209.3) nmol/L, Z=2.06, 2.71, both P<0.05). The serum taurolithocholic acid in both boys and girls were positively correlated with age (r=0.31, 0.32, both P<0.05). The serum chenodeoxycholic acid and glycochenodeoxycholic acid in the boys group were positively correlated with age (r=0.20, 0.23, both P<0.05), whereas the serum tauroursodeoxycholic acid in the girls group was negatively correlated with age (r=-0.27, P<0.05), and the serum cholic acid was positively correlated with age (r=0.34, P<0.05). Conclusions: The total bile acid levels are relatively stable in healthy children in Zhejiang province. However, individual bile acids showed gender differences and were correlated with age.

Sclerotinia Rot of Windflower (Pulsatilla koreana Nakai) Caused by Sclerotinia nivalis in China.

Windflowers (Pulsatilla spp.) are perennial medicinal plants in the family Ranunculaceae with high economic as well as medicinal value in China. It is a commonly used traditional Chinese medicine (1). In 2012, a stem rot disease was observed on windflower (P. koreana Nakai) at flowering stages in fields of Liaoning Province, China. Disease incidence ranged from 10 to 65% (average of 45%) and resulted in approximately 25 to 60% yield loss. The infected area of plants initially takes on a dark green or brown water-soaked appearance and then becomes paler. Soon after, plants turn brown or black and die. If moisture conditions remain conducive, a cottony mycelium cover was observed on the affected area. Later, the mycelium usually produces numerous black sclerotia that were up to 1 cm long in affected plant parts. Diseased tissue were surface sterilized for 1 min in 1% NaOCl (v/v), rinsed with sterilized distilled water, and plated on potato dextrose agar (PDA). Isolates were mesophilic, with an optimum growth temperature of around 20°C. Colonies on PDA were white, with abundant aerial mycelium. Some mycelium in the colony center, especially that submerged in the medium, was light brown. Sclerotia were spherical to subspherical, elongated or fused to form irregular shapes, 2.5 to 9.0 × 2.0 to 6.8 mm. They were tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. These characteristics were consistent with Sclerotinia nivalis (2,3). Two isolates were selected for molecular identification. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (4) and sequenced. The obtained sequences (GenBank Accession Nos. JX206828 and JX424615) showed 99 and 100% homology with the sequences of S. nivalis in GenBank (Accession No. AB516670). To demonstrate pathogenicity, mycelial blocks of the isolates grown on PDA were placed on the base of the stems of ten 1-month-old Pulsatilla koreana plants. The same numbers of control plants were treated with PDA plugs as a control. The inoculated plants were incubated at 25°C with a 12-h photoperiod. After 4 days, the initiation of stem necrosis was observed, and 9 days after inoculation, the plants collapsed and died. Control plants had no symptoms. The same fungus was reisolated from all inoculated plants, satisfying Koch's postulates. To our knowledge, this is the first confirmed report of windflower as a natural host for S. nivalis in China. References: (1) S. C. Bang et al. J. Nat. Prod. 68:268, 2005. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience. 38:227, 1997. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Severe Outbreak of Rust Caused by Coleosporium pulsatillae Detected on Pulsatilla spp. in China.

Windflower (Pulsatilla spp.) is a perennial medicinal plant in the Ranunculaceae with high economic value as well as medicinal value in China. It is a commonly used Chinese herbal medicine. In 2007, two species, Pulsatilla koreana Nakai and P. chinensis Regel, were observed with severe rust symptoms at three locations (Shenyang City, Benxi City, and Fushun County) in Liaoning, China. The diseased area was estimated to be more than 500 ha and the yield was reduced by 30% on average with up to 65% yield losses in some fields. Since its first record in 2007, the disease has been recorded every year in parts of China. A survey of all cultivated fields in August 2011 revealed that 90% of the Pulsatilla plants were heavily infected with rust. Early symptoms on the adaxial leaf surfaces were small, circular, yellow spots that later enlarged, coalesced, and developed necrotic centers. On the abaxial side, numerous yellow rust uredinia were visible. Urediniospores are globose or ellipsoidal, sometimes angular in shape, with a yellowish content, and measured 22.6 to 39.4 × 15.2 to 23.9 µm. Spore walls were coarsely verrucose when examined by light and scanning electron microscopy. Telia and teliospores were observed in mid-August. Hypophyllous telia often formed around uredinial clusters. Telia were 0.3 to 1.1 mm wide and erumpent with numerous teliospores in a compact layer. Teliospores were clavate, oblong to ellipsoidal, 60.2 to 120.8 × 12.1 to 24.4 µm, gelatinous, and one celled. Teliospores were four celled with the division of the protoplast into an internal four-celled basidium. Pathogenicity tests included inoculation of young P. koreana plants by spraying a urediniospore suspension (30,000 spores/ml of deionized water). Inoculated plants were incubated at 25°C for 48 h, and typical rust symptoms and uredinia developed in 10 to 15 days. On the basis of symptomatology, the host, and morphology of uredinial and telia, the fungus was identified as Coleosporium pulsatillae (Str.) Lév (2). The internal transcribed spacer (ITS) region was amplified using ITS1-F and ITS4-B primers (3), and an amplified product of 817 bp, specific for the species C. pulsatillae, was obtained. The sequence was deposited in GenBank (Accession No. JQ029765). Although this pathogen has been mentioned as part of a fungal species survey from China, it was not fully described (4). This pathogen has been reported on Anemone chinensis in Austria, Sibiria (2), and Korea (1). It has not been reported from anywhere else in China. To our knowledge, this is the first fully described record and most severe outbreak of C. pulsatillae on windflower. References: (1) W. D. Cho and H. D. Shin. List of Plant Diseases in Korea. Korean Society of Plant Pathology, Seoul, Korea, 2004. (2) J. B. De-Toni. Sylloge Fungorum 7:754, 1888. (3) M. Gardes et al. Mol. Ecol. 2:113, 1993. (4) F. L. Tai. Sylloge Fungorum Sinicorum. Science Press, Beijing, 1979.

First Report of Sclerotinia Rot of Chinese Atractylodes Caused by Sclerotinia nivalis in China.

Chinese atractylodes (Atractylodes japonica Koidz.ez Kitam.) is a perennial herb in the Compositae family, and is widely distributed in China. The dried rhizomes of the plant are used in traditional Chinese medicine. During the summer of 2011, typical signs and symptoms of Sclerotinia rot were observed on Chinese atractylodes in a production field of Liaoning Province of China. Symptoms were observed in plants at the flowering stage, distributed in patches throughout the rows, and with a disease incidence of approximately 10 to 15%. The lower mature leaves of infected plants first became yellow and wilted, basal stem areas showed a black-brown rot at the same time under conditions of high humidity, and white cottony mycelium formed along the basal stem and soil surfaces. Ultimately, the basal stem and roots rotted and the plants wilted and died quickly. Black, irregular sclerotia (average 0.8 to 6.9 mm in diameter) were also observed within the pith cavity of split stems and rotted roots. The pathogen was isolated from symptomatic tissues and sclerotia, surface disinfested with 2% sodium hypochlorite, and cultured on potato dextrose agar (PDA) (1). The fungus was mesophilic, with an optimum temperature for mycelial growth in culture of about 20°C. Colonies on PDA produced masses of white aerial mycelium, with small white flocci distributed among sclerotia. After 2 weeks, sclerotia 0.5 to 4.5 mm in diameter were produced near the margin in a uniform distribution. Sclerotia were spherical, elongated, or fused to form irregular shapes and tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. DNA sequences of five replicates were obtained using the TianGen DNA secure plant kit. The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and sequenced. BLAST analysis of the 513-bp segment showed high similarity (99%) with a sequence of Sclerotinia nivalis (GenBank Accession No. AB516670). A representative sequence was deposited in GenBank (Accession No. JX294862). The fungus isolated from symptomatic tissues was identified as S. nivalis Saito on the basis of morphological and cultural characteristics (2,3) and ITS sequence analysis. Symptoms were reproduced in the greenhouse by inoculating the basal stem and roots of 15 atractylodes plants at the 7- to 10-leaf stage. Inoculum was prepared by macerating 14-day-old PDA cultures of the fungus in a blender and placing the mixture (approximately 20 g) into the potting medium of each plant. Sterile PDA was used to inoculate the five control plants. Plants were maintained in a greenhouse at 22 to 25°C and about 75% relative humidity. After 7 to 10 days, symptoms were similar to those in the fields. Lower leaves of inoculated plants became yellow and wilted, and infected plants died 2 weeks after inoculation, whereas control plants remained healthy. The pathogen was successfully recovered from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of Sclerotinia rot of Chinese atractylodes. Given its wide host range, S. nivalis has great potential to become an economically important plant pathogen. References: (1) W. G. Kim and W. D. Cho. Mycobiology 30:41, 2002. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience 38:227, 1997.

[Relationship between body mass index and sexual development in Chinese children].

Objective: To investigate the relationship between body mass index (BMI) and sexual development in Chinese children. Methods: A nationwide multicenter and population-based large cross-sectional study was conducted in 13 provinces, autonomous regions and municipalities of China from January 2017 to December 2018. Data on sex, age, height, weight were collected, BMI was calculated and sexual characteristics were analyzed. The subjects were divided into four groups based on age, including ages 3-<6 years, 6-<10 years, 10-<15 years and 15-<18 years. Multiple Logistic regression models were used for evaluating the associations of BMI with sexual development in children. Dichotomous Logistic regression was used to compare the differences in the distribution of early and non-early puberty among normal weight, overweight and obese groups. Curves were drawn to analyze the relationship between the percentage of early puberty and BMI distribution in girls and boys at different Tanner stages. Results: A total of 208 179 healthy children (96 471 girls and 111 708 boys) were enrolled in this study. The OR values of B2, B3 and B4+ in overweight girls were 1.72 (95%CI: 1.56-1.89), 3.19 (95%CI: 2.86-3.57), 7.14 (95%CI: 6.33-8.05) and in obese girls were 2.05 (95%CI: 1.88-2.24), 4.98 (95%CI: 4.49-5.53), 11.21 (95%CI: 9.98-12.59), respectively; while the OR values of G2, G3, G4+ in overweight boys were 1.27 (95%CI: 1.17-1.38), 1.52 (95%CI: 1.36-1.70), 1.88 (95%CI: 1.66-2.14) and in obese boys were 1.27 (95%CI: 1.17-1.37), 1.59 (95%CI: 1.43-1.78), and 1.93 (95%CI: 1.70-2.18) (compared with normal weight Tanner 1 group,all P<0.01). Analysis in different age groups found that OR values of obese girls at B2 stage and boys at G2 stage were 2.02 (95%CI: 1.06-3.86) and 2.32 (95%CI:1.05-5.12) in preschool children aged 3-<6 years, respectively (both P<0.05). And in the age group of 6-10 years, overweight girls had a 5.45-fold risk and obese girls had a 12.54-fold risk of B3 stage compared to girls with normal BMI. Compared with normal weight children, the risk of early puberty was 2.67 times higher in overweight girls, 3.63 times higher in obese girls, and 1.22 times higher in overweight boys, 1.35 times higher in obese boys (all P<0.01). Among the children at each Tanner stages, the percentage of early puberty increased with the increase of BMI, from 5.7% (80/1 397), 16.1% (48/299), 13.8% (27/195) to 25.7% (198/769), 65.1% (209/321), 65.4% (157/240) in girls aged 8-<9, 10-<11 and 11-<12 years, and 6.6% (34/513), 18.7% (51/273), 21.6% (57/264) to 13.3% (96/722), 46.4% (140/302), 47.5% (105/221) in boys aged 9-<10, 12-<13 and 13-<14 years, respectively. Conclusions: BMI is positively correlated with sexual development in both Chinese boys and girls, and the correlation is stronger in girls. Obesity is a risk factor for precocious puberty in preschool children aged 3-<6 years, and 6-<10 years of age is a high risk period for early development in obese girls.

[Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development].

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

[Pseudohypoparathyroidism and GNAS gene defects: clinical evaluation and molecular analysis in 20 children].

Objective: To analyze the patients' clinical and genetic characteristics with pseudohypoparathyroidism (PHP) and investigate the correlation between clinical phenotypes and genotypes. Methods: Twenty PHP patients were ascertained at Children's Hospital Zhejiang University School of Medicine from January 2011 to July 2020. Clinical manifestation, laboratory examination and gene test results were retrospectively analyzed. Results: In these twenty patients, eighteen cases showed resistance to parathyroid hormone (PTH) and thirteen cases had Albright's hereditary osteodystrophy (AHO) phenotype. Gene abnormalities were found in all the twenty PHP patients, which included seven patients with GNAS gene variations (six frameshifts and one missense) and thirteen patients with GNAS gene methylation defects. Moreover, twelve children with both PTH resistance and AHO phenotype were clinically diagnosed as PHP-Ⅰa, meanwhile, seven carried GNAS variations and five had methylation abnormalities with a correct diagnosis of PHP-Ⅰb. Conclusions: Patients with AHO phenotype and PTH resistance may have a high genetic diagnosis rate. Because PHP-Ⅰb clinical phenotype may be similar to PHP-Ⅰa, early genetic detection is required for the differential diagnosis. In addition, children without PTH resistance should also be followed up regularly, which may help the early diagnosis.

[Changing prevalence of overweight and obesity among Chinese children aged 6-15 from 2009-2019].

Objective: To analyze the trends of overweight and obesity prevalence in Chinese children, aged from 6 to 15 years old among 4 provinces and cities from 2009 to 2019. Methods: Reviewed the national multi-center epidemiological survey data of children from the National Key Technology R&D Program of China during the Eleventh Five-Year Plan (2009 to 2010) and the National Key Research and Development Program of China during the Thirteenth Five-Year Plan (2017 to 2019). The participants' data were selected from four provinces,municipalities and autonomous region,including Beijing, Tianjin (Northern region), Zhejiang (Eastern region), and Guangxi (Southern region). Totally 14 597 pairs of 6-15 year-old children were surveyed. According to the body mass index (BMI) and standard deviation score (SDS) of children among different genders, ages, and regions, t test or chi-square test was used to evaluate the changes in overweight and obesity over a 10-year span. Results: Totally 7 721 pairs of boys and 6 876 pairs of girls were collectted in this study, whose mean age was (10.7±2.5) years. In the past 10 years, the overall BMISDS were 0.39±1.24 and 0.36±1.31 and the overall obesity rate were 11.8% (n=1 773) anel 12.5% (n=1 813) of children in the 4 administrative regions did not have statistically significant differences (all P>0.05). However, the overall overweight rate rose from 17.1% (n=2 496) to 19.1% (n=2 781) (χ²=18.657, P<0.01), and the average annual growth rate was 0.20%. The BMISDS in the Eastern region increased from 0.10±1.07 to 0.19±1.22 (t=-4.095, P<0.01), and the overweight rate and obesity rate increased by 3.8% (n=202) and 3.1% (n=169) respectively (both P<0.01); the BMISDS in the Northern region and the obesity rate did not have statistically significant differences(all P>0.05), but the overweight rate rose from 20.5% (n=1 233) to 22.8% (n=1 365) significantly (χ²=7.431, P<0.01); BMISDS in the Southern region was significantly decreased from 0.30±1.19 to 0.09±1.25 (t=1.426, P<0.01), and the rate of obesity decreased from 9.8% (n=315) to7.9% (n=256) (χ²=6.46, P<0.05), the overweight rate was not stafistically significant (P=0.10), respectively. The obesity rate of boys had risen from 16.4% (n=1 265) to 18.2% (1 407) (χ²=8.997, P<0.01) in the past 10 years, and the overweight rate had risen from 18.0% (n=1 393) to 20.5% (n=1 579) (χ²=14.26, P<0.01). The overweight+obesity rate rose from 34.4% (n=2 658) to 38.7% (n=2 986) (χ²=29.859, P<0.01), and the weight problem in the age group of 8 to 11 years was particularly severe (all P<0.01). The obesity rate of girls dropped from 6.8% (n=468) to 5.9% (n=406) (χ²=4.546, P<0.05), the overweight rate rose from 16.0% (n=1 103) to 17.5% (n=1 202) (χ²=5.006, P<0.05), and the overall overweight+obesity rate rose from 22.8% (n=1 571) to 23.4% (n=1 608) (χ²=0.53, P>0.05). Conclusions: The growth rate of obesity among children in China had slowed down from 2009 to 2019, but the overweight rate was still on the rise. The overall base of overweight and obesity population continued to expand. The weight problem of peri-adolescent boys was particularly prominent. The current status of obesity epidemics in different regions, ages, and genders are significantly different and had their own characteristics. It is necessary to establish a personalized prevention and control strategy.

[Survey of height and weight of children and adolescents at different Tanner stages in urban China].

Objective: To investigate the status of height and weight of 3-18-year-old children and adolescents in urban China, and to provide a basis for establishing puberty phase specific curves for age-specific height and age-specific weight. Methods: A cross-sectional survey of 218 185 children and adolescents aged 3-18 years in urban China was conducted by using the method of stratified random cluster sampling from January 2017 to December 2019. The sampling areas included 12 provinces municipalities in China and autonomous regions in total. Data were collected on weight, height, waist circumference, hip circumference and secondary sexual characteristics. The generalized additive model for location, scale, and shape (GAMLSS) was employed to establish percentile reference values and growth curves of height and weight for boys and girls aged 3-18 years. Wilcoxon rank sum test was applied to compare the P50 value of height and weight between children of each Tanner stage and children of the same age ignoring the different puberty phase. Results: The 3rd, 50th, and 97th percentile curves for height and weight for age were developed for boys and girls aged 3-18 years. The 3rd, 50th, and 97th percentile curves for age-specific height and age-specific weight for each puberty phase were developed for boys and girls. Compared with all children ignoring the different puberty phase, boys aged 9 and over and girls aged 7 and over who are at Tanner stage 1 showed shorter height and lighter weight than those of the same age group (all P<0.01), the difference ranges of height at P50 are -4.0 to -0.6 cm for boys, and -4.4 to 0.5 cm for girls; the difference ranges of weight are -4.8 to 0.4 kg for boys, and -4.0 to -0.3 kg for girls; children at Tanner stage 2 & 3 initially were taller and heavier than those of the same age group; and later grew shorter and lighter than those of the same age group, the two sets of curves cross over; boys aged 16 and under and girl aged under 14 who are at Tanner stage 4 were taller and heavier than those of the same age group (all P<0.01), the difference ranges of height at P50 are 0.2 to 10.0 cm for boys, and 0.2 to 9.4 cm for girls; the difference ranges of weight at P50 are 0.7 to 10.9 kg for boys, and 1.0 to 11.2 kg for girls, and the differences showed narrowing trend with age. Conclusion: The puberty phase specific growth curves of age-specific height and age-specific weight for boys and girls aged 3-18 years are established, it is useful for clinical work to evaluate physical development of children at different puberty phases.