Aim To explore the feasibility of the micro- dynamic chromogenic method for quantitative detection of bacterial endotoxin in recombinant novel coronavirus vaccine ( CHO cell).Methods The micro-dynamic color method of Limulus reagent was used to establish a bacterial endotoxin standard curve.The dilution factor was determined through interference pre -experiment, the recoverv rate of the endotoxin added to the test so- J lution was determined, and the interference test to complete the quantitative detection test of the bacterial endotoxin content in the test product was performed, and the results were compared with those of the gel-clot method.Results Hie linear range of the concentration of the standard curve was 0.02 to 2.0 EU • mL 1 , and the regression equation of the standard curve was lgT =-0.302 7 lgC +2.858 7( r = 0.998 9).When recombinant novel coronavirus vaccine ( CHO cell) was cliluted 40 times or below, the micro -dynamic chromogenic reagent did not interfere with the bacterial endotoxin agglutination reaction, and the recovery rate was 50% to 200%.The test results were consistent with the gel- clot method.Conclusions The micro-dynamic chromogenic method can be used for the quantitative detection of bacterial endotoxins in recombinant novel coronavirus vaccine ( CHO cell) with accurate results, high sensitivity, and process monitoring.

Objective : To observe the dynamic expression of recombinant lysyl oxidase like protein 1 ( LOXL1) in the lysine oxidase family in the liver of C57BL/6 mice infected with Schistosomajaponicum and explore its role in hepatic fibrosis． Methods: Mice were infected subcutaneously with cercariae of S．japonicum，and sacrificed with euthanasia in 6，9 and 12 weeks after infection．The sera and liver tissues were collected．The levels of liver fibrosis in mice was dynamically evaluated by HE and Sirius red staining，and the serum transaminases were detected．The dynamic expression levels of collagen type Ⅰ ( Col1) ，LOXL1 and α-smooth muscle actin( α-SMA) in liver tissues were determined respectively by Western blot and qPCR. Finally，the dynamic levels of soluble and insoluble collagens were detected. Results: The result of HE and Sirius red staining showed that hepatic fibrosis levels increased at 6 weeks，peaked at 9 week，and decreased at 12 week in response to S．japonicum infection．Western blot and q-PCR showed that the expression levels of LOXL1，Col1 α1，Col3 α1 and α-SMA was significantly up regulated and reached maximum at the 9th week in response to S．japonicum infection．Soluble collagen protein levels reached maximum at the 9th week．and decreased at 12 week，however insoluble collagen protein levels continued to increase． Conclusion There may be a correlation between LOXL1 and fiber cross-linking in the process of hepatic fibrosis in S．japonicum，and it plays a role in promoting hepatic fibrosis.

ObjectiveTo investigate the efficacy of intensity-modulated radiotherapy (IMRT) in treating large primary hepatocellular carcinoma (LHCC) which is unsuitable for surgery or has poor response to radiofrequency ablation, interventional therapy, and other local treatments, and to identify the prognostic factors for survival. MethodsWe retrospectively analyzed the clinical data of 29 LHCC patients who received IMRT from April 2008 to August 2011. There were five fractions per week and the dose for each fraction was 2 to 6 Gy; the total dose was 50 to 70 Gy. The short-term efficacy and prognosis were observed and analyzed. The Kaplan-Meier method was used to calculate survival rates and the log-rank test was used for survival difference analysis. Multivariate analysis was performed using the Cox regression model. ResultsThe complete remission, partial remission, stable disease, and disease progression rates were 3.57%, 32.14%, 53.57%, and 10.72%, respectively. The overall median progression-free survival (PFS) time was 6.43 months, and the median overall survival (OS) time was 11.43 months. The 1- and 2-year survival rates were 46.79% and 25.23%, respectively. Univariate analysis showed tumor response rate was an independent prognostic factor for PFS. The Cox proportional hazard model suggested the tumor response rate and prescribed dose were the independent prognostic factors for PFS. In addition, the independent prognostic factors for OS included tumor response rate, tumor diameter, and tumor volume. The common acute radiotherapy toxicities included gastrointestinal discomfort, radiation-induced liver damage, and myelosuppression. ConclusionIMRT is a safe and effective option for the LHCC patients who are unsuitable for surgery or in the cases that other local therapies fail.

OBJECTIVE: To explore effects of acupotomy on pain, function, gait and serum inflammatory factors in patients with knee osteoarthritis(KOA). METHODS: From December 2017 to June 2019, 110 patients with KOA were collected and divided into acupotomy group(56 cases) and western medicine group(54 cases) by using random number table method. In acupotomy group, there were 16 males and 40 females, aged from 46 to 74 years old with an average of (62.98±6.68) years old, the course of disease ranged from 1 to 240 months with an average of 24.5(15.25, 33.00) months;were treated with acupotomy on the pain points around knee joint once a week for 3 weeks. In western medicine group, there were 18 males and 36 females, aged from 47 to 73 years old with an average of (64.19±5.98 ) years old;the course of disease ranged from 1 to 220 months with an average of 25.00(13.75, 33.00) months;were took celecoxib capsule orally, 200 mg once a day for 3 weeks. Oxford Knee Score(OKS) was performed before treatment, 3 weeks and 3 months after treatment. Gait kinematics analysis and serum levels of tumor necrosis factor-α(TNF-α) and interleukin-1β (IL-1β) were measured before and after treatment for 3 weeks. RESULTS: All patients were followed up from 6 to 24 months with an average of(15.03±4.55) months. OKS between two groups decreased significantly at 3 weeks and 3 months after treatment(P<0.001). Functional scores and overall scores in acupotomology group were significantly decreased at 3 months compared with 3 weeks after treatment(P<0.001). OKS of acupotomy group were significantly lower than those of western medicine group at 3 weeks and 3 months after treatment(P<0.05). Gait speed, frequency and length between two groups were significantly improved at 3 weeks after treatment(P<0.05). At 3 weeks after treatment, gait freguency of acupotomy group was significantly improved compared with western medicine group(P<0.05). TNF-α and IL-1β were significantly lower in both groups at 3 weeks after treatment than before treatment(P<0.05). At 3 weeks after treatment, level of IL-1 β was lower in western medicine group than in acupotomy group(P<0.05), and difference in TNF-α level was not statistically significant(P>0.05). CONCLUSION Acupotomology of pain points could significantly improve pain, function, gait, and decreased serum inflammatory factors at early to mid stage of KOA patients, in particular, it is superior to non-steroidal anti-inflammatory drugs in terms of knee function recovery and cadence improvement.
Acupuncture Therapy ; Aged ; Anti-Inflammatory Agents, Non-Steroidal/therapeutic use* ; Celecoxib/therapeutic use* ; Female ; Gait ; Humans ; Interleukin-1beta ; Male ; Middle Aged ; Osteoarthritis, Knee/drug therapy* ; Pain/drug therapy* ; Tumor Necrosis Factor-alpha

This paper collects professor ZHOU Mei-sheng's academic thought, "three-phases moxibustion sensation" and expounds its clinical value. Proposed by professor ZHOU, in accordance with the occurrence and development characteristics of the moxibustion propagated sensation, three time phases of moxibustion propagated sensation are divided, i.e. directional conduction phase (the first time phase), effect onset phase (the second time phase), and descending suspension and along-meridian re-transmission phase (the third time phase). In terms of the different characteristics among these three time phases, the clinical therapeutic regimens are designed accordingly. It provides a novel approach to the clinical application of moxibustion.
Acupuncture Therapy/history* ; Meridians ; Moxibustion ; Sensation

OBJECTIVETo study the effects of Polygona-polysaccharose (PSP) on blood glucose level and the mechanism of protection on diabetic rats induced by streptozotocin (STZ).

METHODThe animal model of diabetes was established by injecting STZ (60 mg x kg(-1)) into its abdominal cavity. The amount of water drinking, food intake, urinary volume and body weight were measured at the fourth week of the treatment. The blood samples were drawn to determine the indexes of blood glucose (FBG) and Glycosylated serum protein (GSP) and blood serum insulin (INS). Pancreatic pathology was studied with morphological method and immunohistochemical method. The distribution of apoptotic cells and the expression of Caspase-3 were observed by TUNEL and immunohistochemistry.

RESULTThe levels of FBG, GSP and the amount of water drinking, food intake, urinary volume in the PSP treated groups were obviously lower than those in the model group while INS increased. PSP decreased the rate of apoptotic cells and the level of Caspase-3.

CONCLUSIONPSP can effectivly decrease blood glucose and increase INS. The mechanism may be related with inhibiting islet cell apoptosis and lowering Caspase-3.

Animals ; Apoptosis ; drug effects ; Blood Glucose ; metabolism ; Blood Proteins ; Caspase 3 ; metabolism ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; metabolism ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; chemistry ; Gene Expression Regulation, Enzymologic ; drug effects ; Glycoproteins ; blood ; Hypoglycemic Agents ; pharmacology ; therapeutic use ; Insulin ; blood ; Male ; Polysaccharides ; pharmacology ; therapeutic use ; Rats

OBJECTIVETo evaluate the feasibility and results of closed reduction and fixation for the treatment of Gartland type I and III supracondylar fracture of humerus in children.

METHODSFrom January 2004 to December 2011,110 children with supracondylar fracture of humerus were treated. Among them, 76 patients were boy and 34 patients were girl, ranging in age from 2 to 13 years old, averaged 7 years old. Ninety-eight patients were extension type and 12 patients were flexion type. There were 32 patients with fracture of Gartland type II and 78 patients with fractures of Gartland type III. Thirty-seven patients had distal rotational displacement. Under anesthesia, the upper arm and forearm of the affected limb were treated with contraction for several minutes. The patients were treated with closed manipulative reduction and plaster external fixation while the satisfactory reduction was confirmed by C-arm X-ray. After the reduction, 1 to 2 Kirschner pins were used through the radial side of the skin and plaster external fixation was carried out. Radiographic examination was conducted within 3 to 4 weeks after surgery,and the plaster and Kirschner pins were then removed, accompanied by rehabilitation training of the patients.

RESULTSAll the patients were followed up,and the duration ranged from 3 to 18 months,with a mean of 12 months. All fractures healed within 6 to 8 weeks and the average healing time was 6.9 weeks. No complications such as cubitus varus deformity, neurovascular injury or Volkmann's contracture occurred. According to Flynn evaluation criteria,74 patients got an excellent result, 26 good and 10 fair.

CONCLUSIONThrough carefully reading X-ray films,Gartland type II and III supracondylar fracture of the humerus in children can be treated with closed reduction, plaster external fixation combined with percutaneous pinning for unstable fracture, which is a simple, less-invasive and satisfactory treatment method.

Adolescent ; Casts, Surgical ; Child ; Child, Preschool ; Female ; Fracture Fixation ; methods ; Humans ; Humeral Fractures ; therapy ; Male ; Manipulation, Orthopedic

OBJECTIVETo study the protective effects and mechanisms of astragaloside (AST) and astragalus saponin I (ASI) on the memory impairment in senescent rats treated by glucocorticoid (GC).

METHODY maze test was performed to determine the effects of AST and ASI on memory impairment of hydrocortisone(HC)-induced senescent rats. Using Ca2+ sensitive fluorescent indicator (Furo-2), free intracellular calcium concentration ([Ca2+]i) was measured by double wavelength fluorescence sepectrophotometer in thymocytes and hippocampal neurons induced dexamethasone (DEX). And apoptosis was detected by DNA gel electrophoresis and flow cytometry.

RESULTCompared with HC control, AST and ASI can improve the memory of the senescent rats treated by HC, lower [Ca2+]i and suppress apoptosis of thymocytes and hippocampal neurons induced by DEX.

CONCLUSIONAST and ASI can delay the aging in rats treated by HC, and its mechanism may includ lowering[Ca2+]i and suppressing the apoptosis of thymocytes and hippocampal neurons.

Aging ; drug effects ; metabolism ; pathology ; Animals ; Apoptosis ; drug effects ; Body Weight ; drug effects ; Calcium ; metabolism ; Dexamethasone ; adverse effects ; Female ; Glucocorticoids ; adverse effects ; Hippocampus ; pathology ; Intracellular Space ; drug effects ; metabolism ; Male ; Memory Disorders ; chemically induced ; metabolism ; pathology ; prevention & control ; Neurons ; drug effects ; pathology ; Rats ; Saponins ; pharmacology

ObjectiveTo observe the effect of gastrodin on the steroid regulatory element-binding protein 1c （SREBP1c） signaling pathway in high-fat high-cholesterol diet （HFHC）-induced mice and explore the mechanism of gastrodin in the treatment of non-alcoholic fatty liver disease （NAFLD）. MethodEight-week-old male C57BL/6J mice were used in vivo and divided into the following four groups， with six mice in each group： normal group， gastrodin group （50 mg·kg^-1）， model group， and model + gastrodin group （50 mg·kg^-1）. NAFLD model was established by feeding mice with HFHC for four weeks， and the mice were euthanized and the liver tissues were collected after four weeks. In vitro experiments were performed using Huh7 cells which were divided into five groups， and induced with free fatty acids （FFA， 200 μmol·L^-1， oleic acid-palmitic acid 2∶1） to establish an NAFLD cell model. After 24 h， different concentrations of gastrodin （0， 5， 10， 20， and 40 μmol·L^-1） were added to each group and cultured for another 24 h. Oil red O staining was used to detect lipid accumulation in mouse liver and Huh7 cells. Hematoxylin-eosin （HE） staining was used to observe pathological changes in liver tissue. Levels of serum total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， and triglycerides （TG） were measured using an automatic biochemical analyzer. Relevant assay kits were used to detect liver TC， TG， and FFA levels. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the expression of lipid synthesis-related proteins fatty acid synthase （FASN）， acetyl-CoA carboxylase 1 （ACC1）， and stearoyl-CoA desaturase 1 （SCD1）. ResultCompared with the normal group， the model group showed significantly increased serum TC， LDL-C， and TG levels （P<0.01）， liver TC， TG， and FFA levels （P<0.01）， increased lipid accumulation in Huh7 cells （P<0.01）， and significantly increased expression levels of lipid synthesis-related genes SREBP1c， FASN， ACC1， and SCD1 in mice and Huh7 cells （P<0.01）. Compared with the model group， after gastrodin treatment， the serum TC， LDL-C， and TG levels in mice significantly decreased （P<0.05， P<0.01）， the severity of fatty liver disease improved significantly， liver TC， TG， and FFA levels decreased significantly （P<0.05， P<0.01）， lipid accumulation in Huh7 cells decreased significantly （P<0.05， P<0.01）， the expression levels of lipid synthesis-related genes SREBP1c， FASN， ACC1， and SCD1 in mice and Huh7 cells decreased significantly （P<0.05， P<0.01）. ConclusionGastrodin can reduce hepatic lipid accumulation and blood lipid levels， improve HFHC-induced NAFLD， and its mechanism of action may be related to the regulation of the SREBP1c lipid synthesis-related signaling pathway.

OBJECTIVE: To explore the mechanism of acupuncture plus medication on treatment of Alzheimer's disease (AD). METHODS: Sixty adult SD rats were randomly divided into a normal group, a sham operation group, a model group, an electroacupuncture (EA) group, a gastrodin group and an EA+gastrodin group, 10 rats in each one. The rat model of AD was established by intraperitoneal injection of D-galactose and bilateral hippocampal injection of Aβ1-40. Two weeks after modeling, the rats in the EA group and EA+gastrodin group were treated with EA at "Baihui" (GV 20) "Dazhui" (GV 14) and bilateral "Zusanli" (ST 36), 30 min per treatment, once a day for consecutive 4 weeks. The rats in the gastrodin group and EA+gastrodin group were treated with intraperitoneal injection of gastrodin, once a day for consecutive 4 weeks. The rats in the normal group, model group and sham operation group were not treated. The morphology of hippocampal neurons was observed by using HE staining. The expression of Bcl-2 and Bax in the hippocampal CA1 area was detected by using immunohistochemical method. The expression of Bcl-2 and Bax protein in hippocampus was detected by using Western blot. RESULTS: The HE staining results showed the arrangement of neurons in the hippocampal CA1 area was regular in the normal group and the sham operation group, and the cytoplasm and nucleus were clearly visible. The neurons in the model group were severely damaged; the cell arrangement was not close, and the cell morphology was incomplete. Compared with the model group, the cell morphology of each intervention group was significantly improved. The immunohistochemistry results showed that, compared with the normal group and the sham operation group, the expression of Bcl-2 in the hippocampal CA1 region in the model group was decreased (<0.05), but the expression of Bax was enhanced (<0.05); compared with the model group, the expression of Bcl-2 was increased (all <0.05) and the expression of Bax was decreased (all <0.05) in all intervention group; compared with the EA group or the gastrodin group, the expression of Bcl-2 was enhanced (<0.05) and the expression of Bax was decreased (<0.05) in the EA+gastrodin group. The result of Western blot method was consistent with that of immunohistochemistry method. CONCLUSION EA and gastrodin could significantly inhibit the expression of Bax and up-regulate the expression of Bcl-2, and the combination of EA and gastrodin has the most significant effect. This indicates that EA combined with gastrodin has synergistic effect on inhibiting the apoptosis of neurons in hippocampus in AD rats, which may be one of the mechanisms of EA plus medication on AD lesions.
Alzheimer Disease ; Animals ; Electroacupuncture ; Hippocampus ; Proto-Oncogene Proteins c-bcl-2 ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein