Objective To determine the effect of willed movement on the expression of Nogo-A and Rho-associated kinase (ROCK) in adult rats with cerebral ischemia.Methods Cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion (MCAO) for 2 h,followed by a 24 h reperfusion in 54 adult rats and the degree of their neurological deficit was evaluated using Longa scale.They were then divided randomly into 3 groups,namely the MCAO group,the environmental modification (EM) group,and the willed movement (WM) group.The rats of MCAO group were raised in a regular breeding box,where they could get food and water freely.Meanwhile,those of the other two groups were raised in a homemade box.For the WM group,the water bottle and food were located on the roof of the homemade box.In each group,six rats were killed on day 3,7 and 15 after reperfusion and their neurological deficits were evaluated respectively.Immunohistochemistry assay was employed to examine the expression of Nogo-A and ROCK in the brain tissue around the ischemic foci.Results The rats of the WM group showed lessened neurological deficits on day 15 compared with the model and EM group.Their expression of Nogo-A decreases from(28.92 ± 2.17)/hpf on day 7 to (24.38 ± 2.29)/hpf on day 15 and that of ROCK did from (40.03 ± 2.14)/hpf to (38.08 ± 2.07) / hpf,lower than those of the model and EM group.However,no significant differences were found in the expression of Nogo-A and ROCK between the model group and EM group at any time points.Conclusion Willed movement could promote the functional recovery of neurological deficits in rats with ischemia after reperfusion,which is probably in relation to restrained expression of Nogo-A and Rho-associated in the tissue around the brain ischemic foci.

Thymus has an important significance to the pathogenesis and treatment for myasthenia gravis (MG). Thymoma accounts for 10 to 15% in the MG patients, which is short of localized damage feature so as to make misdiagnosis usually. Using citric acid extracted sacrolemma protein antigen from human skeletal muscle, CAE-Abs in the sera of 103 MG patients were detected by indirect hemoagglutination assay (IHA) and the positive rate was 78.9% (cases of 15/19); and then the CAE-Abs in the sera of 123 MG patients were detected by ELISA and the positive rate'was 82.6% (cases of 19/23). But the positive rate of CAE-Ab in the sera of MG without thymoma was only 0-4.0% (cases of 3/77). This results indicate that CAE-Ab has significant correlativity for MG with thymoma.

Seventy-four patients aged 26-63 yr who had suffered cervicogenic headache for 3 months-21 yr were treated with puked radiofrequency applied to C2 dorsal root ganglion, which is located in the middle of the posterior side of lateral atlantoaxial joint. A trochar was introduced percutaneously under the guidance of X-ray aiming at the target point. As it was inserted through the deep fascia, the stylet was withdrawn and a 10 cm long 22 gauge curved blunt electrode was inserted into the trochar and advanced until the patients felt radiating pain from the point of puncture to occiput. Lateral radiograph was obtained to verify the placement of electrode. The tip of the electrode was usually located in front of spinal canal at the atlantoaxial joint level. Sensory stimulation was performed with 50 Hz and 0.1-0.5 V and the patients could feel radiating pain at occiput. Motor stimulation was performed with 2 Hz and 0.4-1.0 V and regular pulsation of the patient's muscle of occiput could occur. Pulsed radiofrequency was applied at 42 ℃7 for 240 s and was performed twice on each side. VAS scores and disturbances of daily activity, mood and sleep were recorded before operation and at 1 week and 1, 3, 6, 12 and 18 months after pulsed radiofrequency treatment. Complications and recurrence within 12 and 18 months were recorded. Follow-up was lost in 22 patients. VAS scores and disturbances of daily activity, mood and sleep significantly decreased after procedure. All of the patients responded without complications like infection, spinal cord and vertebral artery injury. Some patients had transient occipital neuralgia which was usually relieved within 24 h. The recurrence rate in 12 and 18 months after operation was 19% and 31% respectively.

From November 2003 to May 2010, intrathecal drug delivery system (IDDS) was implanted in 18 patients with chronic intractable pain. Analgesia was provided with morphine. Thirteen patients suffered from late stage cancer and 5 from diseases other than cancer. VAS score was used to measure intensity of pain in all 18patients. QLQ-C30 score was used to evaluate quality of life in cancer patients. The patients were followed up for 3-62 months in 5 non-cancer patients. All 13 cancer patients died at 57 days-10 months after operation. VAS scores were significantly decreased and QLQ-C30 scores increased by intrathecal administration of morphine. Side effects developed in all patients to some extent including nausea, vomiting, constipation, urinary retention, pruritus and over-sedation and vanished in a week. Intrathecal catheter was cut while being pulled out of the needle in 1 patient. Two patients developed low intracranial pressure after operation. Cerebrospinal fluid leakage occurred in 1 patient. One patient developed neuropathic pain in the posterolateral side of right leg.

In this study, the HPLC-UV-MS method for the simultaneous determination of eight active ingredients of Shengxuebao Mixture were developed based on the concept of quality by design（QbD）with a stepwise optimization approach. After the analytical target profile（ATP）had been defined, albiflorin, paeoniflorin, 2, 3, 5, 4'-tetra-hydroxy-stilbene-2-O-β-D-glucopyranoside, specnuezhenide, ecliptasaponin D, emodin, calycosin-7-glucoside, and astragaloside Ⅳ were identified as the indicator components. The resolution and the signal-to-noise ratio of indicator components were then selected as critical method attributes (CMA) for the first step optimization. According to the results collected from fractional factorial design, critical method parameters (CMP) were determined with a multiple linear regression method, which included the amount of acid addition in the mobile phase, temperature, gradient, and wavelength. After that, the amount of acid addition and the wavelength were optimized to improve the resolution and the signal-to-noise ratio of the indicator components. The peak symmetry factors of specnuezhenide and emodin were then set as CMA for the second step optimization. The Box-Behnken designed experiments were conducted. The temperature and gradient were optimized after modelling. The design space were calculated and verified. The optimized analytical method was validated, and the results showed a good precision, accuracy and stability, which means that it can be used for the quantification of the indicator components in Shengxuebao Mixture.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Mass Spectrometry ; Phytochemicals ; analysis ; Reproducibility of Results

To study the preventive effects of double guidewire technique combined with pancreatic duct stenting in preventing post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP). Patients receiving ERCP were divided into the treatment group and the control group by random number table. In the treatment group, double guidewire technique combined with pancreatic duct stenting was applied. In the control group, selective biliary intubation was applied in the conventional way. The intubation time, PEP, hyperamylasemia and bleeding incidence were analyzed between the two groups. A total of 80 patients were enrolled in this study from January 2016 to December 2018. There were 40 cases in the treatment group and 39 cases in the control group. In the treatment group, the mean intubation time was 384±102 seconds. No PEP or bleeding during and after the operation occurred, but hyperamylasemia occurred in 2 cases. In the control group, the mean intubation time was 427±115 seconds. Hyperamylasemia occurred in 6 cases, PEP occurred in 3 cases, and 1 case of intraoperative bleeding happened in the control group. The incidence of PEP [0 VS 7.7%(3/39)]and hyperamylasemia [5.0% (2/40)VS 15.4%(6/39)] were lower in the treatment group (both P<0.05). Double guidewire technique combined with pancreatic duct stenting can successfully perform selective bile duct intubation and effectively prevent PEP.

Objective:To study the safety and efficacy of endoscopic papillary balloon dilation for choledocholithiasis.Methods:A total of 60 patients with choledocholithiasis in Suqian People′s Hospital of Nanjing Drum Tower Hospital Group were included from January 2017 to December 2018 according to the inclusion and exclusion criteria. According to the random number table, the patients were divided into two groups: simple endoscopic papillary balloon dilation group (EPBD group, n=30) and endoscopic papillary sphincterotomy combined with balloon dilation group (ESBD group, n=30). Lithotripsy time, X-ray exposure time, one-time lithotripsy rate, lithotripsy rate, incidence of postoperative acute pancreatitis, intraoperative and postoperative bleeding rates were compared.Results:The time of stone extraction (8.5±2.4 min) in EPBD group was comparable with that of group ESBD (7.8±2.1 min) ( P=0.14). The time of X-ray exposure was 21.8±5.2 min in EPBD group and 19.7±6.3 min in ESBD group ( P=0.11). Stones were extracted at one time in all 60 patients, and no lithotripsy was conducted. The incidences of acute pancreatitis after endoscopic retrograde cholangiopancreatography(ERCP) in the two groups were both 6.67% (2/30). The intraoperative bleeding rates were 3.33% (1/30) and 10.00% (3/30) in EPBD group and ESBD group ( P=0.042), respectively. The rate of postoperative bleeding was 3.33% (1/30) in ESBD group ( P=0.035). No other recent complications occurred in the two groups. Conclusion:Endoscopic papillary balloon dilation alone is safe and effective in the treatment of choledocholithiasis.

Objective:To investigate the inhibitory effects and mechanism of Reyanning mixture （RYN） combined with linezolid （LNZ） against methicillin-resistant Staphylococcus aureus （MRSA） and its biofilm. Method:The minimum inhibitory concentrations （MICs） of RYN and LNZ against MRSA were determined by microdilution assay. The microplate method was used to detect the changes in viable count before and after MRSA administration at four time points （0， 6， 12， 24 h） in the process of biofilm growth. The morphological changes of MRSA after 24 h were observed by scanning electron microscope. Metabonomic technique was applied to analyze the changes in terminal metabolites of endogenous small molecules from MRSA treated by the two drugs at four time points. Result:The MICs of RYN and LNZ were 1/2 of the stock solution concentration and 4 mg·L^-1， respectively. The inhibitory effect of LNZ （2 mg·L^-1） against viable bacteria at 0 h was better than that of 1/16 RYN. At 6， 12， 24 h， 1/16 RYN was superior to LNZ in inhibiting MRSA. The inhibitory effects of RYN combined with LNZ were better than those of RYN or LNZ alone at the four time points. RYN combined with LNZ caused more severe damages to the morphological structure of MRSA biofilm at 24 h than RYN or LNZ alone. Cyclic adenosine monophosphate （cAMP）， adenosine diphosphate （ADP）-D-ribose and 2-methylbutanoyl-coenzyme A （2M-CoA）， as the metabolites related to biofilm formation， were immune to LNZ， but 2M-CoA and ADP-D-ribose were influenced by RYN at 12 h and 24 h. The combined use of RYN and LNZ interfered with the three metabolites at 24 h. L-tryptophan， phenylpyruvic acid， cytidine and sebacic acid were the pharmacometabolic markers of LNZ， and the related biological pathways were phenylalanine， tyrosine and tryptophan biosynthesis and phenylalanine metabolism. Four metabolites such as L-histidine， uric acid， and L-lysine were the pharmacometabolic markers of RYN， with phenylalanine metabolism and aminoacyl-transfer ribonucleic acid （tRNA） biosynthesis confirmed as the related biological pathways. Nine metabolites such as L-tryptophan， L-lysine， and sphingosine-1-phosphate were responsible for the efficacy of RYN combined with LNZ. The related biological pathways involved aminoacyl-tRNA biosynthesis， phenylalanine， tyrosine and tryptophan biosynthesis， novobiocin biosynthesis， and tyrosine metabolism. Conclusion:RYN combined with LNZ better exerts the inhibitory effects against MRSA at each time point of its biofilm formation， which is attributed to cAMP metabolism. The synergistic effect resulted from aminoacyl-tRNA biosynthesis and phenylalanine， tyrosine and tryptophan biosynthesis. RYN combined with LNZ can serve as a potentially effective solution to MRSA infection.

Objective To investigate the effect of Yangxue Rougan pills on a rat model of liver fibrosis induced by multiple factors and the mechanism of action of Yangxue Rougan pills in the treatment of liver fibrosis. Methods A total of 50 male rats were randomly divided into blank control group, multi-factor model group, Fuzheng Huayu capsule group, and high-, middle-, and low-dose Yangxue Rougan pill groups. The rats in the blank control group were given normal water and feed, and those in the other groups were given modified high-fat low-protein diet and 5% alcohol, as well as subcutaneous injection of olive oil solution containing 40% carbon tetrachloride and intraperitoneal injection of pig serum 0.5 mL per rat, twice a week for 12 consecutive weeks. Since week 7, the rats in the high-, middle-, and low-dose Yangxue Rougan pill groups were given Yangxue Rougan pills at a dose of 9.5, 4.75, and 2.38 g/kg, respectively, those in the Fuzheng Huayu capsule group were given Fuzheng Huayu capsules at a dose of 0.75 g/kg, and those in the blank control group and the multi-factor model group were given an equal volume of distilled water by gavage every day for 6 consecutive weeks. The rats were treated at week 12. HE staining and Masson staining were used to observe the degree of liver fibrosis in rats, and PCR and Western blot were used to measure the expression of TGF-β1, Smad3, and Smad7 in the liver. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Dunnett's t -test was used for further comparison between two groups. Results Compared with the blank control group, the multi-factor model group had a severely damaged lobular structure and a significantly higher degree of liver fibrosis, with the formation of pseudolobules with different sizes; compared with the multi-factor model group, the Yangxue Rougan pill groups had a significant improvement in the degree of liver fibrosis, with the most significant therapeutic effect in the high- and middle-dose Yangxue Rougan pill groups. Compared with the blank control group, the multi-factor model group had significant increases in the expression of TGF-β1 and Smad3 and a significant reduction in the expression of Smad7 in liver tissue (all P < 0.05); compared with the multi-factor model group, the Yangxue Rougan pill groups had a significant reduction in the expression of TGF-β1 and a significant increase in the expression of Smad7 (all P < 0.05); compared with the multi-factor model group, the high- and middle-dose Yangxue Rougan pill groups had a significant reduction in the expression of Smad3 (both P < 0.05). Conclusion Yangxue Rougan pills can significantly inhibit liver fibrosis in rats by downregulating the expression of TGF-β1 and Smad3 and upregulating the expression of Smad7, and therefore, the TGF-β1/Smad signaling pathway is one of the mechanisms of action of Yangxue Rougan pills in improving liver fibrosis.

The pharmaceutical manufacturing model is gradually changing from intermittent manufacturing to continuous manufacturing and intelligent manufacturing. This paper briefly reviewed the supervision and research progress in continuous pharmaceutical manufacturing in China and abroad and described the definition and advantages of continuous pharmaceutical manufacturing. The continuous manufacturing of traditional Chinese medicine(TCM) at the current stage was summarized in the following three terms: the enhancement of the continuity of intermittent manufacturing operations, the integration of continuous equipment to improve physical continuity between units, and the application of advanced process control strategies to improve process continuity. To achieve continuous manufacturing of TCM, the corresponding key technologies, such as material property characterization, process modeling and simulation, process analysis technology, and system integration, were analyzed from the process and equipment, respectively. It was proposed that the continuous manufacturing equipment system should have the characteristics of high speed, high response, and high reliability, "three high(H~3)" for short. Considering the characteristics and current situation of TCM manufacturing, based on the two dimensions of product quality control and production efficiency, a maturity assessment model for continuous manufacturing of TCM, consisting of operation continuity, equipment continuity, process continuity, and quality control continuity, was proposed to provide references for the application of continuous manufacturing technology for TCM. The implementation of continuous manufacturing or the application of key continuous manufacturing technologies in TCM can help to systematically integrate advanced pharmaceutical technology elements and promote the uniformity of TCM quality and the improvement of production efficiency.
Medicine, Chinese Traditional ; Reproducibility of Results ; China ; Quality Control ; Pharmaceutical Preparations