The famous classical formulae are epitomes of the clinical practice experience created by doctors in history， as an important symbol of traditional Chinese medicine （TCM） theory and clinical treatment approaches， it has been given a new concept connotation and become one of the breakthroughs for the development of TCM in the new era. Due to the limitations of historical literature and different cognitive perspectives of literature research， there are still common problems in determination of the dosage， original materials， processing and decocting method for the famous classical formulae， which hinder its development and registration process. In the history of more than 2 000 years， famous classical formulae have been developed in the continuous clinical application of practice. This paper systematically reviewed the evolution of the concept and history of the famous classical formulae， and introduced the evolution of the famous classical formulae in terms of name， composition， medicine， dosage， decocting method and clinical functions， including the stability of basic prescription composition， differentiation of drug bases， the progress of processing methods， the characteristics of dosages in different historical periods and the expansion of functions. In view of the research of the key information for famous classical formulae， the paper puts forward the suggestion of paying more attention to the evolution of prescriptions in the past dynasties. In textual research， we should combine the changes of medicinal resources， habits of drug use and production technology of the past dynasties， so as to bridge the ancient and modern. As to the dosage selection， we should be based on the weights and measures of all dynasties and current clinical practice to select rational dosage and decocting method. On the basis of inheriting the essence， follows the ancient but not be bounded by it， and solves the common problems in the textual research of key information from the perspective of history and development.

Objective:To explore the distribution of different subsets of monocyte in peripheral blood of patients with thyroid-associated ophthalmopathy (TAO).Methods:A cross-sectional study was performed.Fifty-nine TAO patients and 30 healthy subjects were recruited continuously in Zhongshan Ophthalmic Center from January 2017 to December 2019.Clinical data of subjects were recorded, and the severity and activity of TAO were graded based on the criteria of NOSPECS and CAS.TAO patients were grouped according to clinical activity of TAO, and the patients were treated by triamcinolone acetonide (TA) injection or methylprednisolone pulse therapy (MPT) accordingly.Peripheral blood of the subjects was collected and monocytes were isolated.The proportion of different monocyte subsets was assayed by a flow cytometry.The differences in distribution of monocyte subsets between TAO group and normal control group, stable TAO group and active TAO group, TA injected group and MPT treated group were compared and analyzed.The study protocol was approved by the Ethics Committee of Zhongshan Ophthalmic Center, Sun Yat-Sen University (No.2014MEKY005), and the written informed consent was obtained from each subject before any medical intervention.Results:The proportion of classical monocyte (CMo) subset in TAO group was (81.77%±5.53)%, which was significantly lower than (84.35±5.83)% in the normal control group ( P=0.034); the proportion of intermediate monocyte (IMo) subset in the TAO group was (10.17±4.19)%, which was significantly higher than (7.69±4.09)% in the normal control group ( P=0.006); no significant difference was found in the proportion of non-classical monocyte (NMo) subset between the two groups ( P=0.892). The proportion of CMo subset in the active TAO group was (77.29±5.80)%, which was significantly lower than (82.64±5.03)% in the stable TAO group ( P<0.01), and the proportion of IMo subset in the active TAO group was (13.79±4.82)%, which was significantly higher than (9.20±3.56)% in the stable TAO group ( P<0.01); no significant change was found in the proportion of NMo subset between the two groups ( P=0.283). There was no difference in the proportion of different TAO subsets before and after TA injection ( P>0.05). In MPT treated group, the proportion of CMo subset in TAO patients was significantly increased and the proportion of IMo subset was significantly decreased (both at P<0.05); there was no significant difference in proportion of NMo subset before and after MPT treatment ( P=0.187). Conclusions:IMo subset is enriched in patients with TAO, and the IMo subset content varies over the disease activity.MPT may inhibit the shift of CMo subset towards IMo subset.

Objective To investigate the effect of gradient heat stress on phagocytosis of hepatic Kupffer cells (KCs) in vitro in rats. Methods Rat Kupffer cells were isolated in vitro and the temperature for gradient heat stress was set at 37, 39, 41 and 43℃. After thermal stimulation, cell injury was detected by PI and Hochest33342 staining. CCK-8 assay was used to investigate difference in cellular proliferation rate over 24h between the groups. Flow cytometry was used to investigate the influence of heat stress on the phagocytosis of KCs. Results Compared to the normal control group, cells in each heat stress group exhibited varying degrees of damage, especially cells in 43℃ group. The ratio of damage cells increased with the increase of heat stress severity (P<0.05). Proliferation assay indicated that the proliferation rate of cells in each heat stress group was significantly decreased in comparison with normal control group 6h after heat stress (P<0.05). After 12h recovery, decrease in proliferation rate was observed only in 43℃ group (P<0.001), and no difference in the rate of proliferation could be observed between the heat stress groups and normal control group after 24h recovery. Flow cytometry showed, that the phagocytosis of KCs decreased in heat stress groups compared with control group, especially in 43℃ group (P<0.05). This phenomenon disappeared after 24h recovery. Conclusion Heat stress can inhibit the phagocytosis of rat liver KCs through its cytotoxic effect on KCs, and subsequently inhibits its proliferative ability. Further investigation of the effect of heat stress on KCs may help understand the pathogenesis of heat stress.

Objective To study the change of the contractile response of human umbilical artery smooth muscle cells (HUASMCs) during the heat stress, and explore the effect of the antioxidant on the changes. Methods HUASMCs were randomly divided into control group, heat stress group, antioxidant preprocessing group. Cells were stimulated by norepinephrine (NE) at a low concentration (0.05mg/L) and at a normal concentration (1.0mg/L) and cultured in the thermostatic water bath (41℃) for 0.5, 1, 1.5 or 2h, respectively. After stimulated by NE, proportion of the cell surface area contraction was measured to reflect the contractile response of each group. Results Compared with control group, regardless of the NE concentration: in heat stress group, contractile response at 1h increased significantly (P<0.05), while at 2h, it was reduced significantly (P<0.05 or 0.01). In the antioxidant preprocessing group, the contractile response was reduced significantly from heat stress to 2h after heat stress (P<0.05 or 0.01). There was no statistically significant difference in contractile response between different NE concentrations in the control group and heat stress group (P>0.05), but in the antioxidant preprocessing group, the contractile response was more significant to the normal NE concentration than to the low NE concentration (P<0.05 or 0.01). Regardless of the NE concentration, the contractile response was lower in the antioxidant preprocessing group than in the heat stress group. Conclusions In the course of heat stress, the contractile response of HUASMCs presents as time-related change. The usage of antioxidant may correct the over-response of HUASMCs to NE in the early heat stress stage, but cannot correct the reduction of the contractile response in the middle and advanced stage.

Objective To investigate the effect of mitogen-activated protein kinases (MAPKs) activation on the heat stressinduced apoptosis of pulmonary microvascular endothelial cells (PMVECs).Methods A mouse model of severe heat stroke was made and TUNEL and immunohistochemistry were employed to detect lung tissue damage.MACS separation was used for isolation of neonatal PMVECs,and TUNEL was utilized to detect the apoptosis of PMVECs.Western blotting was used for determining the MAPKs activation during heat stress recovery (0,2,6h).The monolayer permeability of endothelial cells was detected in terms of transmembrane resistance (TEER) and horseradish peroxidase (HRP).Cells were pretreated with MAPKs activation inhibitors to examine the effect of heat stress on the monolayer cell permeability and apoptosis.Results In mice with severe heat stroke,extensive apoptosis of PMVECs was found in their pulmonary tissues.TUNEL revealed that the number of apoptotic cells increased over time during heat stress recovery period and heat stress could activate MAPKs in PMVECs.Compared with heat stress group,in the cells pretreated with p38 or ERK activation inhibitor PD98059 and SB203580,the monolayer permeability and apoptosis increased while in cells pretreated withJNK inhibitor SP600125,the cellular permeability and apoptosis decreased.Conclusion In mice with severe heat stoke,PMVECs might experience apoptosis and p38 and ERK could inhibit apoptosis while JNK could promote apoptosis.

Objective To study the effects of deoxycytidine (5-aza-2 deoxycytidine, DAC) on DNA Methylation state and expression of mRNA and protein of pl6 gene in human squamous lingual carcinoma SCC-9 cells in vitro. Methods The SCC-9 cells were divided into four groups, group 0, 1, 2 and 3 which processed using three gradients concentration of DAC. The group 0 without DAC was as the control group. Q-MSP was used to detect the state of methylation of the p16 in SCC-9 cells treated by DAC after 48 hours. Real-time fluorescence quantitative PCR was used to detect mRNA expression level changing of the p16 in SCC-9 cells treated by DAC. Immunohistochemical method was used to detect the expression of p16 protein. Results The hypermethylation and non-methylated p16 gene in SCC-9 was mixed with the results of Q-MSP. The results of Real-time PCR showed that mRNA expression of p16 in SCC-9 cells which treated by the different concentration of DAC for 48 hours was higher the control group. And difference was statistically significant (P < 0.05). The high expression of p16 protein was found in the experimental group with immunohistochemical method. Conclusion The p16 gene methylation states of SCC-9 may be suppresses and the recovery of mRNA and protein expression of p16 gene must be prompted by DAC.

Background and purpose:Currently, subjective questionaire is the most frequently used methods to evaluate swallowing dysfunctions after radiotherapy in nasopharyngeal carcinoma patients, while lacking of effective objective examinations. This study aimed to explore effective methods to evaluate swallowing dysfunctions after radiotherapy in nasopharyngeal carcinoma patients, and gain knowledge of the incidence and severity of swallowing dysfunctions. Methods: From Oct. 2013 to Dec. 2013, 128 consecutive outpatients with previously treated nasopharyngeal carcinoma received esophageal barium lfuoroscopy examination at there regularly follow-ups to evaluate swallowing function. Among these patients, 89 were primary treated with intensity modulated radiation therapy (IMRT) and 39 with conventional radiotherapy (CRT). In this study, each patient received esophageal barium lfuoroscopy examination for 3 times with thin, thick and pasty barium and were dynamically observed using X-ray fluoroscopy from front and lateral direction. Swallowing dysfunctions were defined as follows:①The bolus could not be swallowed and blocked in the mouth;②The dilute barium diverted to the glottis or trachea;③Residual barium delayed in the pyriform sinus and vallecula;④The movement of the hyoid bone or epiglottis were restricted;⑤Bolus prolong through the pharynx;⑥Barium slowed down when went though the esophageal entrance. Results:Of the 128 patients, incidence of dysphagia was 60.2%for the entire cohort, 52.8%for IMRT group and 76.9%for CRT group. Incidence of dysphagia for IMRT group was signiifcantly lower than CRT group (P=0.018). Dysphagia incidence within 1 year, 1 to 2 years and more than 2 years after RT were 63.1%, 33.3%and 69.0%, respectively (P=0.019). Conclusion:There was a high incidence of swallowing dysfunction for the nasopharyngeal carcinoma patients treated with radiotherapy and dysphagia incidence decreased when treated with IMRT. Esophageal barium lfuoroscopy examination is objective method to evaluate the incidence and severity of the swallowing dysfunction.

Objective To prepare mouse model with heat stress and determine its pathological changes of the lung and brain during heat stress. Methods BALB/c mouse were randomly (random number) divided into two groups, control group and heat stress group. The animals in the control group were sham- heated at a temperature of ( 25 ± 0.5) ℃ and humidity of (35 ± 5 ) %. The animals of heat stress group were placed in a prewarmed incubator maintained at (35.5 ± 0.5) ℃ and relative humidity of (60 ± 5) %. Rectal temperature (Tc) was monitored, and when Tc respectively reached 39 ℃, 40 ℃ , 41 ℃ and 42 ℃, those study animals were killed. The other animals were removed from the incubator and allowed to cool at an ambient temperature of (25 ±0. 5)℃ and humidity of (35 ±5)% , respectirvely for 12 and 24 hrs when Tc reached 41 ℃ , and for 6 hrs when Tc reached 42 ℃. The lung and brain of all the animals were isolated. Hematoxylin and eosin stain and light microscope were used to detect their pathological changes. Results All the animals displayed uniform response to the heat stress. Low degree of heat stress could induced obviously pathological changes of the lung, progressively greater damage to lung with further congestion of lung matrix, asystematic hemorrhage of alveolar space, abscission of alveolar epithelial cell and disappear of pulmonary alveolus tissue structure were detected with the rise of Tc to 42 ℃. However, absorption of congestion and hemorrhage and recovery of pulmonary alveolus tissue structure could also be seen with cooling at ambient temperature. With low degree of heat stress, the brain only showed moderate edema. Neuronal denaturation and necrosis were detected when Tc reached to 42 ℃. Interestingly, the lesions of brain further aggravated even through cooling treatment after Tc reached to 42 ℃ , but recovery could been observed after cooling treatment followed with Tc of 41 ℃. Conclusions The pathological changes of the lung and brain showed distinctive lesions to heat stress and cooling treatment, and these changes were correlated with the timing and time of cooling treatment, which provide the experimental basis to further study the mechanisms between the heatstroke and multiple organ dysfunction syndrome (MODS).

Objective To evaluate the effectiveness and safety of minimal incision laparotomy assisted endoscopic resection for polyps in small intestine in patients with Peutz-Jeghers syndrome. Methods The clinical data of patients with Peutz-Jeghers syndrome, who underwent minimal incision laparotomy assisted endoscopic resection for polyps in small intestine, were retrospectively studied. The size and number of the endoscopically rosected polyps, and the procedure related complications were documented. Results A total of 812 polyps in small-bowel were resected in 8 patients, in which 384 were smaller than 10mm, 356 with diameter ranging from 11 mm to 30mm, and 72 were larger than 30mm in diameter. Procedure related complications included intestinal dysfunction in 1 patient and abdominal pain in another. No bleeding with a fall in Hb or other severe complications were observed. Conclusion Minimal incision laparotomy assisted endoscopic resectio is a safe and reliable procedure for the treatment of mid-small bowel polyps in patients with Peutz-Jeghers syndrome. It revolutionizes the therapeutic options for polyps in the region of the mid-small bowel and limits the indications for primary surgical management.

OBJECTIVE To observe the effect of the quickread bioindicator. METHODS According to the GB15981-1995 The Measures and the Standards of the Assessment of Disinfection and Sterilization,to control conventionally the(7 high-pressure) steam sterilization cabinets by the quickread-bioindicators. RESULTS Using the effective control method shortened the waiting time by 45 hours,and in-time provided the qualified disinfected supply. CONCLUSIONS The effect of using the quickread-bioindicator resolves the problem of delay of the control result.This is an ideal,scientific,and practical method for effective control of the high pressure steam sterilization.