Following the principles of evidence-based medicine,in accordance with the structure and drafting rules of standardized documents,based on literature research,according to the characteristics of chronic cough in children and issues that need to form a consensus,the TCM Guidelines for Diagnosis and Treatment of Chronic Cough in Children was formulated based on the Delphi method,expert discussion meetings,and public solicitation of opinions.The guideline includes scope of application,terms and definitions,eti-ology and diagnosis,auxiliary examination,treatment,prevention and care.The aim is to clarify the optimal treatment plan of Chinese medicine in the diagnosis and treatment of this disease,and to provide guidance for improving the clinical diagnosis and treatment of chronic cough in children with Chinese medicine.

OBJECTIVES: Ozone is widely applied to treat allergic skin diseases such as eczema, atopic dermatitis, and contact dermatitis. However, the specific mechanism remains unclear. This study aims to investigate the effects of ozonated oil on treating 2,4-dinitrochlorobenzene (DNCB)-induced allergic contact dermatitis (ACD) and the underling mechanisms. METHODS: Besides the blank control (Ctrl) group, all other mice were treated with DNCB to establish an ACD-like mouse model and were randomized into following groups: a model group, a basal oil group, an ozonated oil group, a FcεRI-overexpressed plasmid (FcεRI-OE) group, and a FcεRI empty plasmid (FcεRI-NC) group. The basal oil group and the ozonated oil group were treated with basal oil and ozonated oil, respectively. The FcεRI-OE group and the FcεRI-NC group were intradermally injected 25 µg FcεRI overexpression plasmid and 25 µg FcεRI empty plasmid when treating with ozonated oil, respectively. We recorded skin lesions daily and used reflectance confocal microscope (RCM) to evaluate thickness and inflammatory changes of skin lesions. Hematoxylin-eosin (HE) staining, real-time PCR, RNA-sequencing (RNA-seq), and immunohistochemistry were performed to detct and analyze the skin lesions. RESULTS: Ozonated oil significantly alleviated DNCB-induced ACD-like dermatitis and reduced the expressions of IFN-γ, IL-17A, IL-1β, TNF-α, and other related inflammatory factors (all P<0.05). RNA-seq analysis revealed that ozonated oil significantly inhibited the activation of the DNCB-induced FcεRI/Syk signaling pathway, confirmed by real-time PCR and immunohistochemistry (all P<0.05). Compared with the ozonated oil group and the FcεRI-NC group, the mRNA expression levels of IFN-γ, IL-17A, IL-1β, IL-6, TNF-α, and other inflammatory genes in the FcεRI-OE group were significantly increased (all P<0.05), and the mRNA and protein expression levels of FcεRI and Syk were significantly elevated in the FcεRI-OE group as well (all P<0.05). CONCLUSIONS Ozonated oil significantly improves ACD-like dermatitis and alleviated DNCB-induced ACD-like dermatitis via inhibiting the FcεRI/Syk signaling pathway.
Animals ; Mice ; Dinitrochlorobenzene/metabolism* ; Skin/metabolism* ; Cytokines/metabolism* ; Interleukin-17/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Dermatitis, Allergic Contact/pathology* ; Dermatitis, Atopic/chemically induced* ; Signal Transduction ; RNA, Messenger/metabolism* ; Mice, Inbred BALB C

Moschus chrysogaster (sifanicus) viral hemorrhagic disease (McVHD) is an acute and highly lethal infectious disease caused by Moschus chrysogaster hemorrhagic disease virus (McHDV) whose genome sequence is highly homologous with rabbit hemorrhagic disease virus. To screen the protective antigen of McHDV and set the basis for study of McVHD vaccine, the antigen epitope of major structural protein VP60 of McHDV was analyzed, and the specific primers were designed to obtain three amplified DNA sequences encoding the main antigen epitope of VP60 from McHDV by using RT-PCR. Then the three DNA fragments were sequenced and cloned to prokaryotic expression vector with pET-28a(+) by using overlap extension PCR, and finally the prokaryotic expression plasmid pET-truncated-VP60 was constructed. Subsequently, the pET-truncated-VP60 was transformed into Escherichia coli BL21(DE3), and the recombinant proteins were expressed by IPTG induction. Finally, the expressed protein was purified and applied to immunize that without immunizing with RHD vaccine, then the antiserum titers were evaluated by the hemagglutination inhibition test, and the immune-protective efficacy of the recombinant proteins was observed and analyzed through animal challenge test. The results showed that the multi-epitope DNA fragments of VP60 of McHDV was successfully expressed in the form of inclusion bodies in E. coli, and the relative molecular weight of recombinant proteins is about 45 kDa. After immunized with the recombinant proteins, 100% of New Zealand white rabbits were resistant to attack of McHDV, which indicates efficient immune-protective efficacy of chosen epitope recombinant protein. The study laid a foundation for the development of the new subunit vaccines of McVHD.

Objective To explore the therapeutic effect of mild hypothermia on the inflammatory response, organ function and outcome in perioperative patients with acute Stanford type A aortic dissection (AAAD). Methods From February 2017 to February 2018, 56 patients with AAAD admitted in our department were enrolled and randomly allocated into two groups including a control group and an experimental group. After deep hypothermia circulatory arrest during operation, in the control group (n=28), the patients were rewarmed to normal body temperatures (36 to 37 centigrade degree), and which would be maintained for 24 hours after operation. While in the experimental group (n=28), the patients were rewarmed to mild hypothermia (34 to 35 centigrade degree), and the rest steps were the same to the control group. The thoracic drainage volume and the incidence of shivering at the first 24 hours after operation, inflammatory indicators and organ function during perioperation, and outcomes were compared between the two groups. There were 20 males and 8 females at age of 51.5±8.7 years in the control group, 24 males and 4 females at age of 53.3±11.2 years in the experimental group. Results There was no obvious difference in the basic information and operation information in patients between the two groups. Compared to the control group, at the 24th hour after operation, the level of peripheral blood matrix metalloproteinases (MMPs) was lower than that in the experimental group (P=0.008). In the experimental group, after operation, the awakening time was much shorter (P=0.008), the incidence of bloodstream infection was much lower (P=0.019). While the incidence of delirium, acute kidney injury (AKI), hepatic insufficiency, mechanical ventilation duration, intensive care unit (ICU) stays, or hospital mortality rate showed no statistical difference. And at the first 24 hours after operation, there was no difference in the thoracic drainage volume between the two groups, and no patient suffered from shivering. Conclusion The mild hypothermia therapy is able to shorten the awakening time and reduce the incidence of bloodstream infection after operation in the patients with AAAD, and does not cause the increase of thoracic drainage volume or shivering.

Objective: To investigate the effect of red light combined with hot compress on diabetic peripheral neuropathy (DPN). Methods: From June 2017 to June 2018, 110 patients with DPN admitted to the Department of Endocrinology, Hangzhou Hospital of Traditional Chinese Medicine were selected in the study.The patients were divided into study group (55 cases) and control group (55 cases) according to the random number table method.All patients were given DPN basic care and treatment, with red light in the control group, and red light combined with hot compress in the study group.The motor nerve conduction velocity (MCV) and the sensory nerve conduction velocity (SCV) of the ulnar nerve, median nerve and common peroneal nerve were compared between the two groups before and after treatment.The total scores of the Toronto clinical scoring system (TCSS) were compared between the two groups before and after treatment.The efficacy of the two groups was compared. Results: Before treatment, there were no statistically significant differences in MCV [(40.45±5.33)m/s vs.(40.14±5.08)m/s, t=0.312, P=0.755; (41.15±5.51)m/s vs.(40.86±5.23)m/s, t=0.283, P=0.778; (42.27±5.84)m/s vs.(41.94±5.75)m/s, t=0.299, P=0.766] and SCV [(39.38±4.82)m/s vs.(39.08±4.60)m/s, t=0.334, P=0.739; (40.13±5.45)m/s vs.(39.86±5.15)m/s, t=0.267, P=0.790; (41.18±5.78)m/s vs.(40.89±5.46)m/s, t=0.278, P=0.782] between the ulnar nerve, median nerve and common peroneal nerve in the two groups.After treatment, the ulnar nerve, median nerve and common peroneal nerve of the two groups were treated.The MCV[(48.77±7.25)m/s vs.(44.62±6.30)m/s, t=3.204, P=0.002; (49.35±7.46)m/s vs.(45.36±6.45)m/s, t=3.001, P=0.003; (49.26±7.13)m/s vs.(46.35±6.22)m/s, t=2.281, P=0.025] and SCV[(47.67±6.52)m/s vs.(43.57±5.61)m/s, t=3.535, P=0.001; (47.77±6.63)m/s vs.(44.31±5.14)m/s, t=3.059, P=0.003; (48.33±7.17)m/s vs.(45.12±6.41)m/s, t=2.475, P=0.015] of the two groups were increased, while which of the study group increased more significantly.Before treatment, there was no statistically significant difference in the total scores of TCSS between the two groups [(10.15±1.23)points vs.(10.45±1.51)points, t=1.142, P=0.256]. After treatment, the total scores of TCSS decreased in the two groups, while which of the study group decreased more significantly[(7.22±0.85)points vs.(8.15±0.96)points, t=5.379, P=0.000]. After treatment, the effective rate of the study group was 87.27%, which of the control group was 63.64%, the difference was statistically significant(χ²=8.295, P=0.004). Conclusion The combination of red light and hot compress on DPN has a more prominent clinical effect, which is worthy of wide application.

Objective To investigate the impact of perioperative mild hypothermia on the neurological function and prog-nosis of patients with acute type A aortic dissection. Methods This study enrolled 65 patients with acute aortic dissection un-derwent surgery during the period of February 2017 to February 2018 and randomly divided them into mild hypothermia group and control group. After the process of deep hypothermic circulatory arrest,patients in the mild hypothermia group were re-warmed to 34 ℃ - 35 ℃ and maintained until 24 h after the operation. While,the patients in the control group were rewarmed to 36 ℃ and were treated with routine rewarm therapy. Baseline characteristics were recorded before the operation and neuro-logical and prognosis related indexes were recorded after the operation for all the patients. At the same time,peripheral venous bloods of all the patients were collected preoperatively and at 1、6、 12 and 24 h after the operation. Serum S 100β and neuron-specific enolase(NSE)levels were measured by ELISA kit. Results Compared with the control group,patients in the mild hypothermia group had a significantly shorter recovery time[ 10. 6 h(IQR:7. 6, 19. 1)vs. 25. 8 h(IQR: 13. 3,54. 2),P =0. 007]. At the same time,serum levels of NSE at 1 h and 6 h after operation and serum levels of S 100β levels at 1、6、 12 and 24 h after operation in the mild hypothermia group were significantly lower than those in the control group(P < 0. 05). In addi-tion,the length of stay in the mild hypothermia group was significantly shorter than that in the control group[ 19 days(IQR: 17, 23)vs. 24 days(IQR: 17,28),P = 0. 036]. However,there was no statistically difference in the incidence of delirium and cerebrovascular accidents between the two groups. Conclusion Perioperative mild hypothermia therapy can significantly re-duce brain cell damage in the patients with acute type A aortic dissection and can shorten postoperative recovery time and hospi-talization time,and thus improve the prognosis of patients.

Objective To investigate the value of endoscopic retrograde appendicitis therapy ( ERAT) in the diagnosis and treatment of atypical acute appendicitis. Methods All the 48 patients suspected of atypical acute appendicitis in Jiangsu Province Hospital from January 2015 to December 2016 were randomly divided into ERAT group and conservative treatment group according to the treatment method. The final appendectomy rate of the two groups was analyzed. Results Only 17 of the 24 patients in the ERAT group received endoscopic treatment because of complex conditions or personal wishes, and 16 cases were diagnosed as acute appendicitis. Surgical resection was performed in 5 cases because of recurrence of the disease after ERAT, and the appendectomy rate was 31. 2％ ( 5/16 ) . In the conservative treatment group, all 24 patients were treated with antibiotics. Twenty of them underwent surgical resection with appendectomy rate of 83. 3％ ( 20/24) , and 1 of them had appendiceal perforation. The appendectomy rate of the ERAT group was significantly lower than that of the conservative treatment group (χ2=11. 111, P<0. 05) . Conclusion ERAT has a high diagnostic and therapeutic value for atypical acute appendicitis.

OBJECTIVETo detect potential mutation of TCOF1 gene in a Chinese family affected with Treacher-Collins syndrome.

METHODSClinical data of the patient was collected. The analysis included history taking, clinical examination and genetic testing. All coding regions of the TCOF1 gene were subjected to PCR amplification and Sanger sequencing.

RESULTSA novel mutation c.2261ins G (p.E95X) of the TCOF1 gene was discovered in the patient. The same mutation was not found in his parents and 100 healthy controls.

CONCLUSIONThe c.2261insG (p.E95X) mutation of the TCOF1 gene probably underlies the disease in the patient. Genetic testing can facilitate diagnosis and genetic counseling for families affected with TCS.

Objective To investigate the epidemiological and molecular virological characteristics in HBV-infected patients with copositive HBsAg and anti-HBs.Methods HBV serological markers were analyzed in 52 070 specimens.The epidemiological characteristics of HBsAg and anti-HBs simultaneously positive patients (the experimental group) and HBsAg positive and auti-HBs negative patients (the control group) were compared.The S protein of HBV coding region was amplified by semi-nested PCR and sequenced.The statistical differences between the two groups were compared in different gene regions,genotypes and different clinical diagnosis.Results HBsAg was positive in 20.40％ (10 621/52 070) of all specimens.In the patients with positive HBsAg,2.48％ (263/10 621) was positive anti-HBs.The prevalence of co-positive HBsAg and auti-HBs was higher in aged 0 to 9 years and greater than or equal to 80 years than that in other age,and the prevalence of positive HBsAg and negative anti-HBs was completely opposite.The mutation rate of S protein in the experimental group was significantly higher than that in the control group (1.52％ vs 0.81％,P ＜0.01) with the mutation in the major hydrophilic region (MHR) (1.68％ vs 0.57％,P ＜0.01).The mutation rates of S protein of HBV carriers,chronic hepatitis B (CHB) patients and patients with liver cirrhosis (LC) in the experimental group were significantly higher than those in the control group (1.47％ vs 0.65％,1.28％ vs 0.84％,2.21％ vs 0.44％,P ＜0.05,respectively),except for the patients with hepatocellular carcinoma (HCC) (1.97％ vs 2.21％,P ＞ 0.05).Conclusion Co-positive HBsAg and anti-HBs in HBV-infected patients was more common in HBsAg positive patients aged 0 to 9 years and greater than or equal to 80 years than the others.Coexistence of HBsAg and anti-HBs in HBV-infected patients may relate to immune escape caused by mutation of S protein (mainly MHR).The mutation rates of S protein in the two groups of patients,co-positive HBsAg and anti-HBs and the positive HBsAg combined with negative anti-HBs,were associated with the stage of liver disease.

Objective To investigate the effects and probable mechanism ofShenfu injection on the oxidaitve damage of H2O2-induced PC12 cells.Methods PC12 cells were cultured and exposed to 100μmol/L H2O2 for 1 h to establish the oxidative damage model. The protective effect ofShenfu injection was observed by the cell survival rate measured by colorimetric MTT assay, and the leakage rate of lactic dehydrogense (LDH). Western blot methods were used to detect the expression of NF-κB signaling pathway.Results Compared with the model group,Shenfu injection at 5, 10, 20 ml/L could improve the PC12 cells survival rate (83.11% ± 2.59 %, 87.99% ± 0.59%, 85.26% ± 1.07%vs. 73.82% ± 1.82%;P<0.01 orP<0.05), decrease the LDH leakage rate (32.75% ± 4.10%, 28.52% ± 1.14%, 35.79% ± 1.62%vs. 64.34% ± 3.18%;P<0.01 or P<0.05). Western blot results showed thatShenfu injection could protect the PC12 cells from oxidaitve damage by suppressing the p-p65/p65 (1.30 ± 0.10, 1.17 ± 0.06, 1.37 ± 0.15 vs. 1.70 ± 0.10;P<0.01 orP<0.05), p-IκBα/IκBα (1.07 ± 0.12, 1.00 ± 0.10, 1.03 ± 0.06 vs. 1.17 ± 0.06; P<0.01 orP<0.05).ConclusionShenfu injection has a obvious antioxidant effect on PC12 cells in vitro.