In order to provide a reference basis for the development of relevant compound preparations， this article takes a comprehensive analysis of the usage and dosage of famous classical formulas in Han dynasty from various perspectives， and gives corresponding countermeasures on this basis. Through the comprehensive analysis of the classification and statistics of Zhongjing's medication characteristics， decoction methods， administration and dosage， and combining conversion methods of weights and measures by ancient medical practitioners， along with the dosage and administration of the listed Han dynasty famous classical formulas， it was found that the "Jiangxi method" served as a general guideline for administration according to Zhongjing's original text. This method allowed for flexible dosing based on the conversion of the ancient measurements to modern equivalents［13.8 g per Liang（两）］， ensuring the safe and effective medication of these formulas. After combing， it is found that although the dosage of single medicine is large in famous classical formulas from Han dynasty， the administration is flexible. The crude drug amount per administration serves as the foundational dose， with the frequency of administration adjusted flexibly according to the condition. This dosing approach becomes the key for the rational development of compound formulations of famous classical formulas. Based on the conclusions of the study， it is recommended that when developing compound formulations of famous classical formulas in Han dynasty， the original administration method and dosage should be respected. The original crude drug amount per administration should be considered as the daily foundational dose， with the frequency of administration described within a range（1 to N times per day， where N is the maximum number of administrations as per the original text）. The specific frequency of administration can be adjusted flexibly by clinical practitioners based on the individual condition. This approach should also be adopted in toxicological studies， where the dosage per administration serves as the basis for toxicity research， and the toxicity profile at the maximum administration frequency should be observed， providing guidance on the clinical safety range. Corresponding drug labels should provide information within a range to indicate toxicological risk intervals.

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.

Moutan Cortex and Paeonia Radix are also traditional Chinese herbal medicines. The market is in great demand. However, the market sales of Moutan Cortex and Paeonia Radix have problems such as pesticide residues, excessive heavy metals and degraded quality, which affect their safety and effectiveness. The establishment of a pollution-free and planting system can effectively reduce the pesticide residue and heavy metal content and improve the quality of the medicinal materials while ensuring the production of Moutan Cortex and Paeonia Radix. The article uses field data which based on P.suffruticosa and P.lactiflora cultivation, combined with growth habits and distribution of origin, this research has developed a pollution-free and planting technology system for P.suffruticosa and P.lactiflora. The system includes planting base regionalization, field management, fertilization, pest control and so on, which provides reference for P.suffruticosa and P.lactiflora.

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 observe the effect of different temperatures on endoplasmic reticulum stress, calcium overload, mitochondria and cell damage in pulmonary microvascular endothelial cells (PMVEC) induced by heat stress, and clarify the mechanism of endothelial cell injury in the process of heat stress to provide experimental basis for clinical prevention and treatment of heat stree. Methods Heat stress model of PMVEC cell was set up. Control group cells were incubated at 37℃, 5%CO2, while heat stress group cells were incubated at 39℃, 41℃, 43℃ for 2h, respectively, then further incubated at 37℃, 5%CO2 for 6h. Pretreatment of cells with 20μmol/L BAPTA-AM or 50μmol/L CsA before heat stress at 43℃. The protein levels of p-PERK, PERK p-eIF2a, eIF2a, ATF4 and GRP78 were analyzed by Western blotting. Intracellular Ca2+, mitochondrial membrane potential and the changes in mitochondrial permeability transition pore were investigated by flow cytometry. The change of caspase-3 was detected by Caspase Assay Kit. Millicell-ERS Volt-Ohm Meter and Accessories was used for determining the changes of transepithelium electrical resistance (TER). Results Compared with the control group, with the increase of heat stress temperature (41-43℃), the phosphorylation of p-PERK and p-eIF2a protein and the expressions of ATF4 and GRP78 proteins were gradually activated, intracellular Ca2+ increased, MPTP pore was opened, mitochondrial membrane potential decreased, cell permeability increased and apoptosis occurred, and it was the most obvious in the 43℃ heat stress group, and the difference was statistically significant (P<0.05). Pretreatment with Ca2+ inhibitors promoted the recovery of the MPTP hole, mitochondrial membrane potential and cell permeability, and reduced the occurrence of apoptosis. While pretreatment with the mitochondrial protective agent did not reduce the release of Ca2+, but it could promote the recovery of cell permeability and reduce the occurrence of apoptosis. Conclusion Heat stress activates endoplasmic reticulum stress response, induces intracellular Ca2+ overload mediated cell and mitochondrial damages in PMVEC cells, which may be one of the important mechanisms of endothelial cell injury induced by heat stress.

Objective To investigate the protective effect of heat shock protein (HSP) 70 on the acute lung injury (ALI) of rats with heat stroke.Methods Sixty four rats were randomly (by employing a random number table) assigned into a sham-heated group (Sham group),heat stress group (HS group),and HS plus gluttamine treatment group (HS+GLN group) and HS plus quercet in treatment group (HS+QU group),16 each.All rats were housed in a artificial climate chamber,with the rats in the sham groups exposed to a temperature of 23 ℃ and humidity of 55％ ± 5％,while the rats of HS,HS+GLN and HS+QU groups to an ambient temperature of 39 ℃ and humidity of 65％.During heat stress or sham heating,rectal temperature (Tr),systolic blood pressure (SBP) and pulse rate (PR) were monitored to observe the difference in heat stress response among the groups.The time point at which the SBP started to drop from the peak level was taken as the point of HS onset.At the onset of HS,heat exposure was terminated,then the rats were immediately removed from the chamber,and returned to room temperature.The rats were scarified 0h and 6h after HS onset respectively.After bronchoalveolar lavage fluid (BALF) was collected,the lungs of all animals were harvested for pathological examination of lung injury.The concentrations of IL-1β,TNF-α and IL-6 in BALF and HSP70 in lung homogenate were measured by using an enzyme linked immunosorbent assay kit.Results Compared with HS and HS+QU groups,the rats in HS+GLN group required significantly greater heat load to induce HS (P＜0.001),and had longer survival time span after HS onset.Compared with Sham group,the concentration of HSP70 in lung homogenate in HS group increased in a time-dependent manner (P＜0.001).In comparison with HS group,the concentration of HSP70 in lung homogenate from HS+GLN group was significantly elevated at each time point (P＜0.001),while the treatment with QU significantly inhibited the expression of HSP70 (P＜0.001).The concentration of IL-1β,TNF-α and IL-6 in BALF significantly decreased in HS+GLN group compared with those in HS group and HS+QU group (P＜0.001).The pathological results showed that the lung injury was milder in HS+GLN group,while the opposite in HS+QU group.Conclusion HSP70 could protect HS rats against ALI by enhancing their thermo-tolerance and inhibiting inflammatory response.

Objective To investigate the role of oxidative stress in acute liver injury in a heat stroke model of conscious rats,and to explore its underlying mechanism.Methods Thirty-two rats were randomly (by using a random number table) assigned into a sham-heated control group (Sham group,n=8),a sham-heated group treated with NAC (Sham-NAC group,n=8),a heat stroke group (HS group,n=8) and a heat stoke group treated with NAC (HS-NAC,n=8).Rats were prepared with pre-warm chamber to initiate heat stoke.The change of rectum temperature (Tr),heart rate (HR) and systolic blood pressure (SBP) were monitored,and the time point of HS onset was recorded.Rats were sacrificed 12h after HS onset.ALT,serum TBIL,IL-6,IL-1β,TNF-α,MDA,T-SOD and GSH in the liver homogenates were measured.Liver tissues were harvested for determining the concentration of reactive oxygen species (ROS),neutrophil infiltration and the histological changes.Results During HS onset,no significant differences were observed in Tr,HR,SBP and heat exposure time between HS group and HS-NAC group (P＞0.05).However,the survival time was significantly longer in HS-NAC group than in HS group (P=0.039).12 hours after HS onset,the concentrations of ROS and MDA in the liver homogenates were significantly higher in HS group than in the other groups (P=0.000),while the concentrations of T-SOD and GSH were much lower than in the other groups (P=0.000).The serum concentrations of ALT and TBIL were significantly higher in HS group than in the other groups (P=0.000).Compare with HS group,the pathological injury was alleviated in HS-NAC group (P=0.000).The neutrophil infiltration level and the concentrations of IL-6,IL-1 β and TNF-α in liver tissue were significantly higher in HS group than in HS-NAC group (P=0.000).Conclusion Oxidative stress may play an important role in the pathogenesis of HS liver injury through its cytotoxic effect and by inducing inflammatory responses.

ObjectiveTo investigate the temporal features of renal injury in rats with severe heat stroke (SHS) and their relationship with inflammatory response.Methods Fifty-six male Wistar rats were randomly divided into normal control group and SHS for 0, 2, 6, 24, 48, 72 hours group (SHS-0, 2, 6, 24, 48, 72 h groups), with 8 rats in each group. Rats were placed in an artificial climate chamber [temperature (39.5±0.2)℃, humidity (60±5)%] to induce SHS model, and the criterion for successful model reproduction was the onset of lowering peak systolic blood pressure (SBP). Then the rats were transferred to room temperature (23.0±0.2)℃ after successful reproduction of the model. The rats of normal control group were kept in room temperature of (23.0±0.2)℃. Heart blood and renal tissue samples were harvested, and the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) were determined by automatic biochemistry analyzer. The levels of myeloperoxidase (MPO), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in renal tissue specimens were determined by enzyme linked immunosorbent assay (ELISA). The changes in histopathology in kidney were observed with light microscopy, and Paller scores were used to assess the degree of renal injury.Results Compared with normal control group, the levels of SCr and BUN in serum, and MPO, TNF-α and IL-6 in the renal tissue homogenate were significantly increased in SHS-6 h group [SCr (μmol/L): 174.0±27.0 vs.68.0±11.3, BUN (mmol/L): 12.6±2.3 vs. 4.3±1.2, MPO: (203.0±38.0)% vs. (100.0±1.4)%, TNF-α: (121.0±16.0)% vs. (100.0±1.4)%, IL-6: (118.0±19.0)% vs. (100.0±1.3)%, allP< 0.05], and they peaked at 24 hours [SCr (μmol/L): 489.0±96.0 vs. 68.0±11.3, BUN (mmol/L): 19.3±5.7 vs. 4.3±1.2, MPO: (511.0±41.0)% vs. (100.0± 1.4)%, TNF-α: (399.0±47.0)% vs. (100.0±1.4)%, IL-6: (473.0±56.0)% vs. (100.0±1.3)%, allP< 0.01], then declined to the normal levels at 72 hours. Under light microscopy, tissue edema and necrosis of renal tubules were found, and leukocyte infiltration was found to be most profuse at 24 hours, then they returned to normal levels at 72 hours. Paller scores in SHS-6 h group were significantly higher than those of the normal control group (75.45±9.70 vs. 14.23±3.26,P< 0.01), and it peaked at 24 hours (186.00±14.25 vs. 14.23±3.26,P< 0.01), followed by a gradual lowering, back to normal level at 72 hours.ConclusionThe results suggest that progressive renal damage occurred in the rats with SHS within 24 hours, and it was accompanied with elevated levels of MPO, TNF-α and IL-6 in the kidney homogenate, suggesting that inhibition of neutrophil activation and the release of IL-6, TNF-α may protect the SHS associated renal injury.

Objective To evaluate diagnostic value of Sonoclot in the diagnosis of heat stroke complicated with disseminated intravascular coagulation (DIC). Methods 43 patients with heat stroke and coagulation dysfunction were retrospectively included and divided into 2 groups according to scores by International Society of Thrombosis and Hemostasis (ISTH): DIC dominant group (22 case) and DIC non-dominant group (21case). Regular coagulation tests , routine blood test , D-dimer and Sonoclot tests were performed at admission and their clinical data were compared. Logistic regression analysis was applied to evaluate the relationships between DIC occurrence and Sonoclot parameters. ROC curves were used to evaluate diagnostic value of Sonoclot for the patients with DIC and heat stroke. Results There were no differences in age, sex, central temperature and total hospital stay between the 2 groups except ICU stay, DIC symptoms, outcome, ISTH scores and APACHEⅡscores (P <0.05). ACT and CR correlated with the occurrence of DIC (P < 0.05). The AUC of ACT and ACT combined with CR were 0.854 and 0.877 respectively. The specificity of ACT in predicting DIC was 69.2%with the sensitivity of 90.3%. When combined with CR, both the specificity and the sensitivity were increased to 80.2% and 93.5%, respectively. Conclusions Sonoclot can predict DIC quickly and is effective in the diagnosis of heat stroke patients with DIC.

Objective To investigate the effect of different intervention times on the efficacy of catheter balloon dilatation therapy for cricopharyngea] achalasia after stroke.Methods Forty-eight cases diagnosed as cricopharyngeal achalasia were divided into 4 groups,groups A,B,C,and D according to the time of intervention from the onset (i.e.0.5,0.5-1,1-3,and 3-6 months),with 12 cases in each group.A 14# urethral catheter was inserted into the esophagus and water was injected into the balloon to make it turgid.The catheter was then pulled upwards through the stricture in the esophagus to dilatate the cricopharygeal muscle.The patients also received low frequency electrical stimulation combined with dysphagia rehabilitation behavioral therapy once daily.The treatment end point was either when the patient resumed an oral diet or after 6 weeks of treatment.The effects of the treatment were evaluated in terms of the number of times the dilatation therapy was administered,the pharyngeal transit duration,swallowing function and fluoroscopic examination before and after treatment.Results Groups A,B and C required significantly fewer treatment sessions than group D.Groups A and B required significantly fewer sessions than group C,but the difference between groups A and B was not statistically significant.After treatment,there were 11 patients can eat water,liquid,paste and solid food both in groups A and B.But the number of patients who can eat the4 traits of food was 9,9,10,9 in group C and 6,6,7,5 in group D respectively.Swallowing function in groups A,B and C was adjudged,on average,significantly better than in group D.Compared with group C,swallowing function in groups A and B was also significantly improved,but the difference between groups A and B was not statistically significant.The pharyngeal transit duration in all 4 groups was shorter than before treatment,but the duration in groups A,B and C was significantly shorter than in group D.Compared with group C,the pharyngeal transit duration in groups A and B was significantly shorter,but the difference between groups A and B was again not statistically significant.Fluoroscopy showed the efficiency in groups A and B to be over 91 ％.In group C it was 83％ and in group D 58％.The difference between group D and groups A,B and C was significant.The efficiency in groups A and B was significantly better than in group C,but the difference between groups A and B again was not statistically significant.Conclusions Early intervention using catheter balloon dilatation therapy can facilitate the improvement of swallowing function in patients with cricopharyngeal achalasia after stroke and improve their quality of life.