Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.

Objective: To investigate the epidemiological characteristics of 45 norovirus clusters in Baoshan District, Shanghai Municipality from 2019 to 2021, so as to provide the evidence for improving the management of norovirus clusters. Methods: The epidemiological characteristics and laboratory testing of 45 norovirus clusters were collected in Baoshan District from 2019 to 2021, and the temporal distribution, population distribution, distribution of places of norovirus clusters, clinical manifestations, laboratory testing results, origin of norovirus infection and transmission routes of norovirus clusters were analyzed using a descriptive epidemiological method. Results: A total of 45 norovirus clusters involving 29 379 individuals were reported in Baoshan District from 2019 to 2021, and 435 cases with norovirus infections were identified, with an average attack rate of 1.48%. Norovirus infection most often occurred from November to April of the following year, and the infection predominantly occurred among students (408 cases, 93.79%). There were 25 norovirus clusters in kindergartens (55.56%), 18 clusters in primary and middle schools (40.00%), one cluster in a social care facility and one cluster in a geriatric ward. Vomiting was the predominant clinical symptom (416 cases, 95.63%). The positive rate of norovirus was 37.35% in 597 samples. There were 40 norovirus clusters caused by the norovirus GⅡ genotype (88.89%), and the median duration of the norovirus cluster was 6 days (interquartile range, 1 day). In addition, 82.22% of the norovirus clusters reported to centers for disease control and prevention within 1 day,and 88.89% of the clusters were transmitted through human to human transmission or vomitus of cases. Conclusion The norovirus clusters predominantly occur in winter/spring and among students in Baoshan District from 2019 to 2021. Kindergartens and primary and middle schools are main places of norovirus infections, and the norovirus GⅡ genotype is the main type of pathogens. Human-to-human transmission and exposure to patients' vomitus are the main routes of transmission.

To study the impact of five different origin processing methods, namely natural drying, drying in baking shop, drying by microwave heating, drying in drum and drying with sulphur fumigation, on the quality of Lonicerae Japonicae Flos from Donghai cultivation base in Jiangsu Province, with the contents of chlorogenic acid and galuteolin and the similarity in HPLC fingerprints as the evaluation indicators. The results showed that different origin processing methods had significant impact on the content of chlorogenic acid and the similarity in HPLC fingerprints, but with no significant difference on the content of galuteolin. By means of drying by microwave heating and drying in drum, the samples showed higher contents of chlorogenic acid, respectively 3.67% and 3.39%. The similarities of HPLC fingerprints were 0.815 and 0.793, respectively. By means of the drying in baking shop and the drying with sulphur fumigation, the contents of chlorogenic acid in the samples were 2. 87% and 2. 53% , respectively. The similarities of HPLC fingerprints were 0.964 and 0.765, respectively. The lowest content of chlorogenic acid in naturally dried samples was 1.92%. The similarity of HPLC fingerprints was 0.940. According to the findings as well as the internal control standards for Lonicerae Japonicae Flos herbs of Jiangsu Kanion Pharmaceutical Co. , Ltd. , the optimum processing method for Lonicerae Japonicae Flos from Donghai cultivation base was the drying in baking shop. This study provided a theoretical basis for determining the processing method for Lonicerae Japonicae Flos from Donghai cultivation base of Jiangsu Province.
China ; Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Lonicera ; chemistry ; growth & development ; Quality Control

Objective To study the treatment of adolescent hyperthyroidism. Methods Retrospective analysis of the clinical data of the adolescent hyperthyroidism treated by surgery was made in our hospital from January 1990 to December 1998. Results In this series, there was no death, with 3 cases transient postoperative hypocalcemia and 1 had throat spasm and physoia. All the 4 cases recovered after treatment. 78 cases(85.7%) were followed up for 0.5～9 years. Of them, all were in good condition except 2 being recurred. Conclusions Operative treatment for adolescent hyperthyroidism is a safe, quick and effictive method, but the operation indications should be good controlled.

Objective: To prove the clinical therapeutic effect of acute and chronic bronchial asthma with Chuankezhi(CKZ) Injection. Methods: 40 cases of bronchial asthma, including inpatients and outpatients were selected, divided into the attack period group (20) and the relief period group (20). The former was intramuscularly injected CKZ injection 4mL, twice a day, 14 days being a course of treatment. The latter was intramuscularly injected CKZ injection 4mL, twice a day for 15～30 days and 2mL, every other day for 31～90 days; 90 days being a course of treatment. The symptoms and signs, such as asthma, cough, expectroation, pulmonary wheeze were observed. and pulmonary function, absolute eosinophilic granulocyte count and immunogobulin E were measured before and after treatment. Result: By the use of CKZ injection to treat bronchial asthma, the total effective rate amounted to 85% and obvious difference before and after treatment in the two groups ( P

OBJECTIVE: To explore the mechanism of Baisuifang Granule in treating cognitive malfunction after cerebral infarction. METHODS: One hundred and sixty patients with cerebral infarction were divided randomly into two groups. Eighty patients were treated with Baisuifang Granule and 80 with nimodipine for two months. Clinical observation and laboratory examinations were performed for Mini-Mental State Examination (MMSE), clinical symptoms, Chinese Stroke Scale (CSS), hemorrheological indexes and fibrinogen before and after the treatment. RESULTS: Baisuifang Granule could improve MMSE, reduce the scores of clinical symptoms and CSS, and meliorate the blood rheology. The total effective rate for clinical symptoms in the Baisuifang treated group accounted to 76.25%, with statistical difference comparing to 58.75% of nimodipine treated group (P<0.05). There was significant difference in symptom integral, CSS and whole blood viscosity at the high shear rate, respectively (P<0.01). CONCLUSION: Baisuifang Granule is an effective Chinese medicine for treating cognitive malfunction after cerebral infarction.

Objective To understand the association between people under impaired glucose regulation (IGR) with poor quality of sleep and the occurrence of diabetes.Methods Based on a cohort of 1136 persons with IGR,the present study would include information on the duration of sleep and severity of insomnia.The cohort was followed for two years and the outcomes of impaired glucose regulation were summarized.Poisson regression was employed to analyze the relationship between the quality of sleep and the occurrence of diabetes.Results After the confounders were adjusted,when compared with 7-9 h sleep duration,the less than 7 h sleep duration showed a RR value of 1.77 (P＜0.001) while more than 9 h sleep duration had a RR value of 1.45 (P=0.067).When compared data from people having less than 14 scores of the Insomnia Severity Index (ISI),those people with more than 14 scores had a RR value of 1.58 (P＜0.001).Conclusion Causal relationship between the quality of sleep and the occurrence of diabetes did exist among the IGR population while the poor quality of sleep might increase the risk of diabetes at the end of the two-year following-up program.

Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).

Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.

Objective To investigate the efficacy of Ilizarov technique in treatment of Gustilo type Ⅲ B and C of large tibia bone defect combined with soft tissue defect. Methods Thirty cases with large tibia bone defect combined with soft tissue defect in our hospital from January 2016 to December 2018 were selected including 25 cases with type Ⅲ B and 5 cases with type Ⅲ C,who were all open tibia fractures and treated with Ilizarov technique. Results All patients were followed up for 10 to 18 months with an average of 15 months. The bone defects were reconstructed, the difference between the length of the affected limb and the healthy side was less than 2 cm. The fracture was healed and the wound was closed. One case was with paralysis of common peroneal nerve and recoveried after 3 months. Twenty-two cases recovered without extra surgery, five cases restored with skin graft and three cases covered the wound by rotating skin flap. Conclusion Ilizarov technique is one of effective ways to treat Gustilo Ⅲ type B and type C of large defect combined with soft tissue defect.