Understanding the research methods for drug protein targets is crucial for the development of new drugs, clinical applications of drugs, drug mechanisms, and the pathogenesis of diseases. Cellular thermal shift assay (CETSA), a target research method without modification, has been widely used since its development. Now, there are various CETSA-based technology combinations, such as mass spectrometry-based cellular thermal shift assay (MS-CETSA), isothermal dose response-cellular thermal shift assay (ITDR-CETSA), amplified luminescent proximity homogeneous assay-cellular thermal shift assay (Alpha-CETSA), etc., which combine their respective advantages and further expand the application scope of CETSA. These technologies are suitable for the entire drug development chain, from drug screening to monitoring the target binding and off-target toxicity of drugs in patients. Based on the author's research experience, this paper reviews the principles of CETSA and related binding technologies, their application in target discovery, and the progress of data processing and analysis in recent years, aiming to provide reference and reference for the further application of CETSA.

Objective: To explore the short-term efficacy of fenestrated atrial septal defect (ASD) occulders in the treatment of pulmonary arterial hypertension (PAH). Methods: Thirty-six healthy dogs were divided into the balloon atrial septostomy (BAS)+fenestrated ASD occulders group (n=12), BAS group (n=12) and non-septostomy group (n=12). PAH was induced by intra-atrial injection of dehydrogenized monocrotaline (1.5 mg/kg) in all dogs. Animals in the BAS+fenestrated ASD occulders group underwent atrial septal puncture and fenestrated ASD occulders implantation. Animals in the BAS group underwent balloon atrial septostomy. The non-septostomy group received no surgical intervention. The hemodynamic indexes and blood N-terminal pro-B-type natriuretic peptide (NT-proBNP) of dogs were measured before modeling, 2 months after modeling, 1, 3, and 6 months after surgery, respectively. Echocardiography was performed to observe the patency of the shunt and atrial septostomy of the dogs in the BAS+fenestrated ASD occulders group and BAS group at 1, 3, and 6 months after surgery. Three dogs were sacrificed in each group at 1, 3, and 6 months after surgery, respectively. Atrial septal tissue and fenestrated ASD occulders were removed to observe the patency and endothelialization of the device. Lung tissues were obtained for hematoxylin-eosin (HE) staining to observe the inflammatory cells infiltration and the thickening and narrowing of the pulmonary arterials. Results: Among 36 dogs, 2 dogs died within 24 hours after modeling, and 34 dogs were assigned to BAS+fenestrated ASD occulders group (n=12), BAS group (n=11), and non-septostomy group (n=11). Compared with BAS group, the average right atrial pressure (mRAP) and NT-proBNP of dogs in the BAS+fenestrated ASD occulders group were significantly reduced at 3 months after surgery (P<0.05), and the cardiac output (CO) was significantly increased at 6 months after surgery, arterial oxygen saturation (SaO₂) was also significantly reduced (P<0.05). Compared with non-septostomy group, dogs in the BAS+fenestrated ASD occulders group had significantly lower mRAP and NT-proBNP at 1, 3, and 6 months after surgery (P<0.05), and higher CO and lower SaO₂ at 6 months after surgery (P<0.05). Compared with the non-septostomy group, the dogs in the BAS group had significantly lower mRAP and NT-proBNP at 1 month after surgery (P<0.05), and there was no significant difference on mRAP and NT-proBNP at 3 and 6 months after surgery (P>0.05). Echocardiography showed that there was a minimal right-to-left shunt in the atrial septum in the BAS group at 1 month after the surgery, and the ostomy was closed in all the dogs in the BAS group at 3 months after the surgery. There was still a clear right-to-left shunt in the dogs of BAS+fenestrated ASD occulders group. The shunt was well formed and satisfactory endothelialization was observed at 1, 3 and 6 months after surgery. The results of HE staining showed that the pulmonary arterials were significantly thickened, stenosis and collapse occurred in the non-septostomy group. Pulmonary microvascular stenosis and inflammatory cell infiltration in the pulmonary arterials were observed in the non-septostomy group. Pulmonary arterial histological results were comparable between BAS+fenestrated ASD occulders group and non-septostomy group at 6 months after surgery . Conclusions: The fenestrated ASD occulder has the advantage of maintaining the open fistula hole for a longer time compared with simple balloon dilation. The fenestrated ASD occulder can improve cardiac function, and it is safe and feasible to treat PAH in this animal model.
Animals ; Atrial Septum/surgery* ; Cardiac Catheterization/methods* ; Dogs ; Familial Primary Pulmonary Hypertension ; Heart Septal Defects, Atrial/surgery* ; Hypertension, Pulmonary ; Pulmonary Arterial Hypertension

The teaching effect of "process management and evaluation" was assessed in resident standardization training plan in acupuncture-moxibustion department of hospital for postgraduates of non-acupuncture-moxibustion speciality. A total of 120 postgraduates of non-acupuncture-moxibustion speciality participating in resident standardization training were randomized into an observation group (60 cases) and a control group (60 cases, 1 case dropped off). In the control group, the conventional training mode was used. In the observation group, the "process management and evaluation" was adopted, in which, the syllabus was refined, various teaching modes were cooperated and the summary was conducted once a week. The training results were evaluated at the end of 1-month shift test and questionnaire was issued in all of the postgraduates of the two groups. In the observation group, the score for theory and the score of each of the items for technical ability, named differentiation and treatment, technical manipulation and physician-patient communication, as well as the total score were all higher than the control group successively (
Acupuncture ; Acupuncture Therapy ; Hospitals ; Humans ; Moxibustion ; Reference Standards

BACKGROUND: Neck pain caused by cervical spondylosis (CS) is a chronic pain condition, with an increasingly high incidence in the general population. Electroacupuncture is a common analgesic modality that has been widely applied in neck pain treatment. However, current electroacupuncture instruments used in the clinic have low intelligence levels and obscure parameter standards. We here designed this study for assessing the effect and safety of a new, intelligent electroacupuncture instrument, the CX-DZ-II, in treating neck pain. METHODS: The present study is a prospective, two-center, randomized, controlled, open-label, non-inferiority trial for CX-DZ-II on treating neck pain caused by CS. Totally 160 eligible patients will be included in this trial and randomly assigned to an experimental group and a control group in a 1:1 ratio. A semi-standard acupoint selection strategy will be employed. In the experimental group, selected acupoints will be stimulated by CX-DZ-II. Electroacupuncture treatment will be accomplished by a pre-existing electroacupuncture instrument in the control group. The duration of treatment will be 2 weeks. The primary outcome is the change of Visual Analog Scale (VAS) score after one course of treatment. The secondary outcomes include the VAS scores after each treatment, the responder rate, drug-usage rate of non-steroidal antipyretic analgesics, the rate of adverse events occurrence, and the performance of instrument. DISCUSSION This study will evaluate the effect and safety of the CX-DZ-II intelligent electroacupuncture therapeutic instrument in comparison with a pre-existing non-intelligent instrument in the treatment of neck pain caused by CS. The results will hopefully demonstrate a more optimal electroacupuncture instrument for the treatment of neck pain. (Trial registration No. gov NCT03005301).
Adolescent ; Adult ; Aged ; Electroacupuncture ; instrumentation ; methods ; Equivalence Trials as Topic ; Female ; Humans ; Male ; Middle Aged ; Multicenter Studies as Topic ; Neck Pain ; therapy ; Pain Measurement ; Prospective Studies ; Spondylosis ; therapy ; Young Adult

Objective:To construct oxaliplatin (L-OHP) drug-resistant cell line HCT-116/L-OHP in human colon cancer, in order to observe the reversal effect of curcumin (cur) on its drug resistance, and preliminarily explore the possible drug resistance mechanism. Method:The concentration gradient increasing method was used to gradually increase the L-OHP concentration of HCT-116 in parental colon cancer cells, and the cell line HCT-116/L-OHP resistant to L-OHP was established. The cytotoxicity of L-OHP and curcumin to HCT-116 and HCT-116/L-OHP cells was detected by cell counting kit-8(CCK-8) method to observe whether curative resistance could be reversed. Western blot was used to detect the expressions of drug-resistance-related proteins. Real-time PCR was used to detect changes in related genes. Result:Human colon cancer cell line resistant to L-OHP were successfully established and named as HCT-116/L-OHP, with a drug resistance index of 12.6.Compared with HCT-116 cell lines, the expression levels of resected and repaired cross complementation gene 1 (ERCC1) protein and gene in HCT-116/L-OHP cell lines were significantly increased (P<0.01). The expressions of B-cell lymphoma-2 (Bcl-2), glutathione-s-transferase-consciousness (GST-consciousness), multidrug resistance-associated protein (MRP), P-glycoprotein (P-gp) and apoptotic inhibitory gene (Survivin) also increased significantly (P<0.01). After the treatment with different concentrations of curcumin (5,10,20,30,40 μmol·L^-1), the expression of ERCC1 decreased (P<0.01), and the expressions of Bcl-2,GST-π,MRP,P-gp,Survivin also decreased to different degrees (P<0.05). Conclusion:HCT-116/L-OHP cell lines have a stable drug resistance, and its drug resistance mechanism may be up-regulated with the expression of ERCC1, which leads to the up-regulation of Bcl-2,GST-π,MRP,P-gp,Survivin and other related proteins, and enables tumor cells to acquire drug resistance. Curcumin can reverse the drug resistance of HCT-116/L-OHP, and its mechanism may be to reduce the expression of ERCC1, thereby down-regulating the expressions of Bcl-2,GST-π,MRP,P-gp,Survivin and other drug-resistant related genes and proteins, and increase the sensitivity of tumor to L-OHP, so as to reverse the drug resistance of tumor cells.

Objective: To investigate the effect of Rhei Radix et Rhizoma on gastrointestinal function of normal rats and explore its therapeutic mechanism,and compare the difference of this herb before and after simmering. Method: Healthy SD rats were randomly divided into 7 groups,including the blank group,the high,medium and low dose groups of raw Rhei Radix et Rhizoma(1.5,3.0,6.0 g·kg^-1·d^-1) and the high,medium and low dose groups of simmered Rhei Radix et Rhizoma(1.5,3.0,6.0 g·kg^-1·d^-1),eight rats in each group were continuous intragastric administration for 10 days,the effect of Rhei Radix et Rhizoma on gastrointestinal function of rats were evaluated by body weight,food intake,gastric remnant rate,small intestine propelling rate,stomach weight coefficient and gastrointestinal tissue morphology;enzyme-linked immunosorbent assay(ELISA) was used to detect contents of motilin(MTL) in rat plasma and gastrin(GAS),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10) in rat serum,the mechanism of effect of Rhei Radix et Rhizoma on gastrointestinal function was explored and the effect of Rhei Radix et Rhizoma on gastrointestinal function of normal rats before and after simmering was compared. Result: In each administration group,weight growth of rats was slowed down,gastric emptying and intestinal propulsion were inhibited,inflammatory reaction was triggered,and gastric mucosal injury was increased.Compared with the blank group, except for the low-dose and medium-dose groups of simmered Rhei Radix et Rhizoma,stomach weight coefficient,gastric remnant rate and content of TNF-α of rats were significantly increased(P<0.05,P<0.01), and the content of IL-10 was significantly reduced, in addition to the low-dose group of simmered Rhei Radix et Rhizoma,the weight,small intestine propelling rate,contents of GAS and MTL of rats were significantly decreased(P<0.05,P<0.01).Compared with the isodose raw Rhei Radix et Rhizoma group,the weight,small intestine propelling rate,contents of GAS,IL-10 and MTL were all increased with different degrees, the gastric weight coefficient,gastric remnant rate and content of TNF-α of rats were all decreased to different degrees. Conclusion: Gastrointestinal dysfunction caused by Rhei Radix et Rhizoma is related to inhibiting the expression of gastrointestinal hormones and promoting the expression of inflammatory cytokines, in addition,the effect of "stomach injured by bitterness and cold property" of Rhei Radix et Rhizoma is weakened after simmering,which provides scientific basis for clinical rational use of Rhei Radix et Rhizoma.

Objective To investigate the effect of CYP46A1 on the pathogenesis of Alzheimer's disease.Methods Recombinant lentiviral vectors which including anthropogenic CYP46A1 were injected into bilateral hippocampus of 3-monthold male 5XFAD transgenic mice,while empty vectors were injected into the corresponding position of the control group.After two months,the ability of learning and memory were tested by Morris water maze and T maze experiments,and amyloid plaque and inflammatory infiltration in the brain were detected by immunohistochemical staining and ELISA respectively.Results Compared with the control group,CYP46A1 virus injection significantly increased the CYP46A1 mRNA and protein expression in hippocampus.In addition,CYP46A1 overexpression significantly decreased the latency to find the platform in Morris water maze test and increased the correct rate to choose in T maze test.Aβ immunohistochemical staining and plaques area statistics demonstrated that the amyloid plaque area of hippocampus in CYP46A1 overexpression mice was significantly reduced,and there was a significantly decrease of hippocampal astrocytes expression by means of GFAP staining.Furthermore,hippocampal CYP46A1 overexpression significantly decreased the expression level of Aβ40,Aβ42,IL-1β and TNF-α,while compare with the control group.Conclusion CYP46A1 overexpression in hippocampus can promote the cognitive impairment,as well as ameliorate the brain inflammatory infiltration in 5XFAD transgenic mice,suggesting that CYP46A1 has anti-Alzheimer's disease like effects.

We summarized our experience in transurethral seminal vesiculoscopy (TSV) for recurrent hemospermia by introducing surgical techniques, intraoperative findings, and treatment outcomes. TSV was performed in 419 patients with an initial diagnosis of persistent hemospermia at Shanghai Changhai Hospital (Shanghai, China) from May 2007 to November 2015. TSV was successfully performed in 381 cases (90.9%). Hemospermia was alleviated or disappeared in 324 (85.0%) patients by 3 months after surgery. Common intraoperative manifestations were bleeding, obstruction or stenosis, mucosal lesions, and calculus. Endoscopic presentation of the ejaculatory duct orifice and the verumontanum was categorized into four types, including 8 (1.9%), 32 (7.6%), 341 (81.4%), and 38 (9.1%) cases in Types A, B, C, and D, respectively. TSV is an effective and safe procedure in the management of seminal tract disorders. This study may help other surgeons to become familiar with and improve this procedure. However, further multicentric clinical trials are warranted to validate these findings.
Adult ; Ejaculatory Ducts/surgery* ; Endoscopy/methods* ; Hemospermia/surgery* ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Seminal Vesicles/surgery* ; Tomography, X-Ray Computed ; Treatment Outcome ; Urethra/surgery*

Objective To develop a HPLC-MS/MS method for the determination of Pregabalin in epileptic patients.Methods Analysis was performed on a Diamonsil C18column (4.6 mm × 150 mm,5 μm)eluted with methanol and water (0.05％ formic acid) in a gradient program.The flow rate was 1 mL · min-1,the injection volume was 20 μL,and the column temperature was 30 ℃.The multiple-reaction monitoring scanning (MRM) was employed for the quantification with switching electrospray ion source polarity in negative mode.The ion spray voltage was set at 5500 V and the turbo spray temperature was maintained at 650 ℃.The specificity,calibration curve,limit of detection,precision,recovery and stability of the method were investigated.20 patients with epilepsy were selected,and the serum concentration of pregabalin was determined by HPLC-MS/MS after oral administration of 75 mg pregabalin for one hour.Results Pregabalin showed a good linearity in the ranges of 13.76-5270.00 ng · mL-1,the lower limit of quantification was 0.65 ng · mL-1.The average recoveries varied from 94.7％-106.2％.The inter and intra-day RSD were less than 15％.Pregabalin was proved to be stable under study station.The concentration of pregabalin in epileptic patients ranged from 1.76 to 2.78 μg · mL-1.Conclusion The method is simple,rapid,and sensitive,which could be used for the determination of pregabalin in epileptic patients.

Objective To evaluate the effect of tranexamic acid on the blood loss and coagulation function after liver resection in patients with hepatocellular carcinoma (HCC).Methods Seventy patients with hepatectomy who underwent liver resection were randomly divided into control group and treatment group,each group 35 cases.Control group was given 0.9％ NaCl injection 10 mg · kg-1.Treatment group was given tranexamic acid 10 mg · kg-1.Two groups were used at half an hour before operation.The surgical bleeding volume and postoperative coagulation function between two groups were compared.Results The operation time,time of hilar occlusion in treatment group were (157.98 ±22.36),(30.14 ±5.64) min,had no significant difference with those in control group,which were (154.98 ± 23.65),(28.79 ± 5.79) min (P ＞ 0.05).The surgical blood transfusion,bleeding volume,infusion of erythrocyte suspension in treatment group were (879.65 ± 59.36) mL,(768.97 ± 45.65) mL,(1.36 ± 0.59) U,had significant differences with those in control group,which were (1054.91 ±61.32) mL,(869.64 ±46.69) mL,(2.01 ±0.65) U (all P ＜0.05).The percentage of activated thromboplastin (APTT),prothrombin time (PT),platelet (PLT) in treatment group at 24 h after operation were (41.56 ±5.46)s,(13.48 ±3.16)s,(167.89 ±35.87) × 109/L,had no significant difference with those in control group,which were (43.19 ± 5.69) s,(14.59 ± 2.49) s,(162.35 ± 41.59) × 109/L (all P ＞0.05).The fibrinogen (FIB) in treatment group and control group were (3.27 ±0.56),(3.01 ±0.59)g · L-1,with significant difference (P ＜ 0.05).There was no adverse drug reactions in two groups during the treatment.Conclusion Giving methotrexate injection to hepatocarcinoma patients before liver resection can reduce the amount of surgical bleeding,and improve the patients' postoperative coagulation function.