Objective To understand the molecular epidemiological characteristics of Norovirus outbreaks and the genome evolution of Norovirus epidemic strains in Hainan Province from 2020 to 2022.Methods The information and samples have been collected from the norovirus outbreaks from 2020 to 2022.Norovirus was detected by using the real-time PCR in these samples,then the detected sequences were amplified the analyzed.The Norovirus se-quences of 8 strains had been amplified and analyzed.Results From 2020 to 2022,39 gastroenteritis outbreaks were reported,and 25 outbreaks caused by Norovirus which mainly occurred in childcare institutions and schools(20/25,80％).The Norovirus outbreaks were mainly concentrated in counties around Haikou(northeast),which including Ding'an(5 cases),Wenchang(4 cases),Chengmai(4 cases),and Lingao(3 cases);following by western regions which included Baisha(2 cases),Ledong(2 cases),and Dongfang(3 cases).1 case was in Wanning in the southeast.Among individuals aged 2-17,the positive proportion of Norovirus in males was higher than that in females.Among individuals aged over 55,the proportion of Norovirus positive in females was higher than that in males.The gender of positive samples among individuals aged 18-40 was related to their profession.According to RT-PCR typing and sequencing,GⅡ group Norovirus were classified in13 outbreaks.There were 4 genotypes detected.GⅡ.2[P1 6]was the main epidemic strain with 60％(9/13),and the other three genotypes were GⅡ.4 Sydney[P31](15.4％,2/13)GⅡ.4 Sydney[P16](7.7％,1/13)and GⅡ.3[P12](7.7％,1/13).Further genic analysis of 8 Norovirus strains showed that all of them were still in the same branch as the previ-ous strain,and all exhibited a certain amount of amino acid variation.Conclusion Norovirus is the main pathogen of gastroenteritis outbreaks in Hainan province,and the main epidemic strain is GⅡ.2[P16].It is necessary to continue to strengthen the monitoring that provides scientific evidence for the prevention and control of norovirus out-breaks in Hainan region.

Clinicians need to focus on various points in the diagnosis and treatment of insomnia.This article prescribed the treatment protocol based on the unique features,such as insomnia in the elderly,women experiencing specific physiologi-cal periods,children insomnia,insomnia in sleep-breathing disorder patients,insomnia in patients with chronic liver and kidney dysfunction.It pro-vides some reference for clinicians while they make decision on diagnosis,differentiation and treat-ment methods.

Objective:To clarify the effects of microwave radiation on anxiety-like behavior, the histomorphology of the secondary visual cortex, and calcium activity in neurons.Methods:36 C57BL/6N mice were selected and divided into control group and microwave radiation group according to the random number table method. In the simple behavioral testing, there were 8 mice in the control group and 7 mice in the radiation group. Combining fiber optic recording with behavioral experiments, there were 8 mice in the control group and 7 mice in the radiation group. Hematoxylin-eosin (HE) staining was conducted with 3 mice in each group. A high-power microwave simulated source in the X-band with a center frequency of 9.875 GHz and an average power density of 12 mW/cm 2 was used to irradiate the mice for 15 minutes, establishing a microwave radiation animal model. Then, anxiety-like behavior changes in the radiation group were identified using the open-field and elevated plus maze (EPM) tests. The effects of microwave radiation on the histomorphology of the secondary visual cortex were investigated using HE staining and optical microscopy. Based on the genetically encoded calcium imaging technique, as well as optical fiber recording combined with behavioral paradigms in the open field and the EPM, the changes of calcium activity in neurons in the V2M region of the secondary visual cortex were detected. Results:Compared to the control group, the radiation group showed a significant decrease in the frequency of exploring the central region of the open field and the open arm of the EPM ( t = 2.24, 3.10, P < 0.05). Furthermore, the radiation group exhibited the degeneration and apoptosis of some neurons in the secondary visual cortex, primarily manifested as pyknosis and deep staining, cell body shrinkage, and the slightly widening of perivascular space. Fiber optic recordings and behavioral experiments indicated that compared to the control group, mice in the radiation group exhibited significantly increased calcium activities in neurons of the secondary visual cortex when exploring the central region of the open field ( t = -2.75, P < 0.05) or the open arm of the EPM ( t = -2.77, -3.41, P < 0.05) compared to those before radiation after microwave exposure. Conclusions:Microwave radiation can induce anxiety-like behaviors and histopathological changes in the secondary visual cortex. Increased calcium activity in neurons of the secondary visual cortex is proved to be an important mechanism underlying the changes in anxiety-like behavior due to microwave radiation.

Objective:To explore the effect of chronic incomplete sleep deprivation (SD) on ovarian reserve function in mice and its potential mechanisms.Methods:Sixteen 7-8 week old female C57BL/6 SPF mice were randomly divided into control and SD groups ( n=8 per group) after one week of acclimatization. The mice in the SD group were treated with SD from 7:00 to 10:00 using a deprivation rod for a total of 40 d. Before the first day and the last day of the experiment, vaginal smears were collected daily for 9 d to record and evaluate the estrous cycle. On the last day of the experiment, serum levels of estradiol, progesterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and melatonin (MT) were measured by enzyme-linked immunosorbent assay (ELISA). The ovaries were dissected to calculate the ovarian index and count the number of primordial follicles by hematoxylin-eosin (HE) staining. Cardiac perfusion was performed to get the brains and the expression of c-Fos protein was observed in the suprachiasmatic nucleus (SCN) by immunohistochemistry. Results:Compared with control group, SD group had a tendency of estrous cycle disorders. Hormone levels of estradiol [(20.19±3.67) ng/L], progesterone [(2.88±0.53) μg/L], and FSH [(13.42±2.36) U/L] in the SD group were significantly lower than those in control group [(24.66±2.15) ng/L, P=0.010; (3.43±0.49) μg/L, P=0.049; (17.01±1.49) U/L, P=0.003]. Hormone levels of LH and MT in the SD group were lower than those in control group, but without statistical significances (all P>0.05). There was no significant change in the number of primordial follicles between the two groups ( P>0.05). However, the oocyte morphology was poor, the granulosa cells were disorderedly arranged, the number of atretic follicles was decreased, and ovarian fibrosis was obvious in the SD group. Immunohistochemical staining showed an upregulation of c-Fos protein expression in the SCN of the SD group. Conclusion:Continuous 3 h SD for 40 d impairs ovarian reserve function in mice, possibly related to excessive activation of the SCN.

Objective:To explore the effect of chronic incomplete sleep deprivation (SD) on ovarian reserve function in mice and its potential mechanisms.Methods:Sixteen 7-8 week old female C57BL/6 SPF mice were randomly divided into control and SD groups ( n=8 per group) after one week of acclimatization. The mice in the SD group were treated with SD from 7:00 to 10:00 using a deprivation rod for a total of 40 d. Before the first day and the last day of the experiment, vaginal smears were collected daily for 9 d to record and evaluate the estrous cycle. On the last day of the experiment, serum levels of estradiol, progesterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and melatonin (MT) were measured by enzyme-linked immunosorbent assay (ELISA). The ovaries were dissected to calculate the ovarian index and count the number of primordial follicles by hematoxylin-eosin (HE) staining. Cardiac perfusion was performed to get the brains and the expression of c-Fos protein was observed in the suprachiasmatic nucleus (SCN) by immunohistochemistry. Results:Compared with control group, SD group had a tendency of estrous cycle disorders. Hormone levels of estradiol [(20.19±3.67) ng/L], progesterone [(2.88±0.53) μg/L], and FSH [(13.42±2.36) U/L] in the SD group were significantly lower than those in control group [(24.66±2.15) ng/L, P=0.010; (3.43±0.49) μg/L, P=0.049; (17.01±1.49) U/L, P=0.003]. Hormone levels of LH and MT in the SD group were lower than those in control group, but without statistical significances (all P>0.05). There was no significant change in the number of primordial follicles between the two groups ( P>0.05). However, the oocyte morphology was poor, the granulosa cells were disorderedly arranged, the number of atretic follicles was decreased, and ovarian fibrosis was obvious in the SD group. Immunohistochemical staining showed an upregulation of c-Fos protein expression in the SCN of the SD group. Conclusion:Continuous 3 h SD for 40 d impairs ovarian reserve function in mice, possibly related to excessive activation of the SCN.

Objective:To study the therapeutic effect Tetrandrine (TET) on striatal injury caused by microwave radiation and underlying mechanism.Methods:C57BL/6N mice were randomly divided into blank control group (C), radiation control group (R), TET group (TET) and TET combined with radiation group (TET+ R). The mice of radiation group were exposed to 2.856 GHz 8 mW/cm2 microwave on whole-body for 15 min. TET (60 mg/kg) was injected intraperitoneally once a day for 3 consecutive days. The TET structure was verified by ultraviolet spectrophotometry. The open field experiment was used to detect the change of anxiety in mice. Histopathological and ultrastructural changes of the striatum were observed by light microscopy and transmission electron microscopy (TMT). Quantitative real-time PCR (qPCR) was used to detect gene expression changes of voltage-gated calcium channel (VGCC) subtype in the striatum.Results:The open field experiments showed that the time and distance of mice to explore the central region after microwave radiation were significantly lower than that before radiation ( t=4.60, 5.18, P<0.01), and the TET administration significantly improved these changes ( F=1.43, 4.37, P < 0.05). 7 d after microwave radiation, some neuronal nuclei in the striatum of mice contracted and could be stained deeply, which was more obvious in the globus pallidus area. The partial neuronal apoptosis, swelling and cavitation of glial cell mitochondria, blurring of synaptic gaps, and widening of perivascular gaps in the striatum were observed by TMT. The above lesions were significantly rescued after TET administration. But both microwave radiation and TET administration had no significant effect on the gene expressions of striatal VGCC ( P > 0.05). Conclusions:TET has a therapeutic effect on anxiety-like behavior and structural damage of striatum caused by microwave radiation, which is independent of the expression of striatal VGCC genes.

Objective:To investigate the risk factors of moderate to severe pain in patients with non-small cell lung cancer within 3 days after lobectomy.Methods:The clinical data of 297 patients with non-small cell lung cancer who underwent lobectomy in the Department of Thoracic Surgery, Sun Yat-sen University Cancer Center from December 2020 to June 2021 were retrospectively analyzed. A numerical rating scale was used to score the most severe pain within 3 days after surgery. Pain score ≥ 4 was defined as moderate to severe pain. The risk factors for moderate to severe pain were analyzed by binary Logistic regression. General linear model repeated measures and linear mixed models were used to analyze the trend of risk factors influencing postoperative pain with time.Results:The incidence of moderate to severe pain was 34.2% (102/297), 59.8% (178/297), 66.4% (198/297), and 28.2% (84/297) on days 0, 1, 2, and 3 after surgery respectively. The risk for moderate to severe pain was significantly higher in patients undergoing thoracotomy than patients undergoing thoracoscopic surgery on days 1 ( OR = 1.99, P = 0.009), 2 ( OR = 3.08, P < 0.001), and 3 ( OR = 3.88, P < 0.001) after surgery. However, the risk for moderate to severe pain in patients undergoing thoracotomy was slightly, but not significantly, higher than that in patients undergoing thoracoscopic surgery ( OR = 1.53, P = 0.087). The risk for moderate to severe pain was higher in female patients than male patients on day 2 ( OR = 1.62, P = 0.077), and in particular on day 3 after surgery ( OR = 2.39, P = 0.002). Prophylactic use of parecoxib significantly reduced the risk of moderate to severe pain on day 0 ( OR = 0.32, P = 0.004), 1 ( OR = 0.20, P < 0.001), 2 ( OR = 0.36, P < 0.001) and 3 ( OR = 0.56, P = 0.047). Conclusion:The incidence of moderate to severe pain on days 1 and 2 after lobectomy was relatively high in patients with non-small cell lung cancer. Patients undergoing thoracotomy have a higher risk of moderate to severe pain than those who underwent thoracoscopic surgery. Female patients have a higher risk for moderate to severe pain on days 2 and 3 after surgery than male patients. Prophylactic use of parecoxib can decrease the risk for moderate to severe pain in patients with non-small cell lung cancer.

OBJECTIVE To investigate the pharmacodynamic effects of the Citri Grandis Exocarpium extract on anorexia model immature rats.METHODS Ten immature rats were randomly selected as the control group and given regular diet， while anorexia model was induced in the remaining 50 immature rats using the etiological simulation method. The immature model rats were divided into the model group， Jianwei xiaoshi tablet group （0.65 g/kg）， Citri Grandis Exocarpium extract low-dose， medium- dose and high-dose groups （0.5， 1.0， 2.0 g/kg）. After 14 d of continuous intragastric administration， the body mass and food intake of rats in each group were observed， and the gastric acid concentration， pepsinase activity， serum levels of gastrin （GAS）， motilin （MTL） and gastric inhibitory peptide （GIP） were measured. The histopathological changes in the gastric tissue were observed.RESULTS Compared with control group， the food intake， gastric acid concentration， pepsin activity， the levels of GAS and MTL were significantly decreased and GIP level was significantly increased in the immature rats of model group （P＜0.05）； gastric histopathological examination showed significant gastric mucosal congestion and loosening and detachment of epithelial cells. Compared with model group， all administration groups showed different degrees of improvement in the above indexes and decreased gastric histopathological damage after drug intervention.CONCLUSIONS Citri Grandis Exocarpium extract improves gastric motility and protects gastric mucosa in the immature rats by regulating the concentration of gastric acid， pepsin activity， the levels of GAS， MTL and GIP， thus improving anorexia in immature rats.

OBJECTIVE: To explore the molecular pathogenesis for a pedigree affected with hereditary coagulation factor XII (FXII) deficiency. METHODS: Potential variant of the F12 gene was analyzed by PCR and Sanger sequencing. Expression plasmids were constructed by site-directed mutagenesis based on the wild-type and transiently transfected into 293T cells. FXII:C and FXII:Ag of the expression products were determined in the supernatant and cell lysate. Western blotting was used to verify the identify of the protein. RESULTS: Gene sequencing revealed that the proband has carried 46TT genetype and heterozygous p.Glu502Lys variants in exon 13, and a heterozygous p.Gly542Ser variant in exon 14 of the F12 gene. Transfection experiment suggested that the FXII:C and FXII:Ag of p.Glu502Lys variant in the supernatant were 28% and 24%, compared with the wild-type (100%) and FXII:Ag of cell lysates was 39% compared to the wild-type (100%). The FXII:C and FXII:Ag of p. Gly542Ser variant in the supernatant were 32% and 17% and the FXII:Ag of cell lysates was 59%. CONCLUSION The 46TT genetype, p.Glu502Lys and p.Gly542Ser variants of the F12 gene probably underlie the low FXII level in the proband. As shown by in vitro experiment, the p.Glu502Lys and p.Gly542Ser variants can both inhibit the synthesis and secrection of the FXII protein.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Heterozygote ; Humans ; Pedigree

OBJECTIVE: To analyze the phenotype and genetic basis for a pedigree affected with hereditary coagulation factor XI deficiency. METHODS: Activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), FXI activity (FXI:C) and the antigen of FXI (FXI:Ag) were determined for the proband and members from his pedigree. Sanger sequencing was used to analyze all exons, exon-intronic boundaries, as well as the 5'- and 3'- untranslated regions of the F11 gene. Suspected variants were verified in her family members and confirmed by reverse sequencing. The impact of the variants on the protein function was predicted by using PolyPhen-2 and SIFT software. The protein structure and amino acid interaction were analyzed by using Swiss-PdbViewer. RESULTS: The APTT, FXI:C and FXI:Ag of the proband and her sister were significantly reduced to 73.0 s, 10.0%, 15.0% and 87.1 s, 2.0% and 11.5%, respectively. APTT of some family members was slightly prolonged, and FXI:C and FXI:Ag also decreased to various extents. DNA sequencing revealed that the proband and her sister have carried compound heterozygous variants of c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) respectively in exons 7 and 9 of the F11 gene. Her father, sister and daughter were heterozygous for the c.738G>A (p.Trp228stop) variant, while her mother and nephew were heterozygous for the c.938G>T (p.Ser295Ile). Both PolyPhen-2 and SIFT predicted that the p.Ser295Ile variant is likely to be deleterious and can affect the protein function. Modeling analysis indicated that the p.Ser295Ile variant may lead to disruption of a hydrogen bond, resulting in alteration of protein structure and instability. CONCLUSION The compound heterozygous c.738G>A (p.Trp228stop) and c.938G>T (p.Ser295Ile) variants of the F11 gene probably underlie the decreased FXI level in this pedigree.
Factor XI Deficiency ; genetics ; Female ; Genetic Variation ; Heterozygote ; Humans ; Mutation ; Pedigree