Effects of Zuogui Pill (左归丸) on Wnt Singal Transduction in Rats with Glucocorticoid-induced Osteoporosis

Objective: To reveal the mechanism of Zuogui Pill (左归丸) in treatment of glucocorticoid-induced osteoporosis from the angle of the Wnt signal transduction pathway and to provide further experimental evidence for expounding the scientific connotation of the kidney dominating the bones in TCM. Methods: Forty-two male Wistar rats were selected and randomly divided into three groups, control group (n=12), model group (n=15) and Zuogui Pill group (n=15). Form the beginning, The rats were injected dexamethasone for eight weeks to make the model of osteoporosis, and the Zuogui Pill were administered intragastrically to the rats of Zuogui Pill group for eight weeks. The relative morphological parameters were measured in the undecalcified tibial slices. And the protein expression levels of Wnt1, LRP-5 and β-catenin in rat tibial osteoblasts (OB) and bone marrow stromal cells (BMC) were detected by immunohistochemistry. Results: Compared with the control group, TBV% and TFS% decreased significantly, while TRS% increased significantly, and the protein expression of Wnt1, LRP-5 and β-catenin in OB and BMC decreased significantly in the model group. And compared with the model group, TBV% and TFS% increased significantly, and expression levels of Wnt1, LRP-5 and β-catenin proteins increased significantly in the Zuogui pill group. Conclusion: Zuogui Pill can prevent and treat glucocorticoid-induced osteoporosis in rats by up-regulating the expression of the key signal molecules Wnt1, LRP-5 and β-catenin in Wnt signal transduction pathway.     

Research on the Mechanism of Zuogui Pill(左归丸)and Yougui Pill(右归丸)in Promoting Axonal Regeneration in Model Rats of Autoimmune Encephalomyelitis

<正>Objective:To study the molecular mechanism of Zuogui Pill(左归丸,ZGP) and Yougui Pill(右归丸,YGP) on axonal regeneration in rats with experimental autoimmune encephalomyelitis(EAE).Methods: EAE rat model was established by bilateral rear pedes subcutaneous injection of antigen made by mixing myelin basic protein(MBP) and complete Freud's adjuvant(CFA) in the volume ratio of 1:1.The pathological changes of axonal injury and regeneration in the brain and the spinal cord were observed on the 14th(the acute stage) and the 28th day(the remission stage) after modeling,with hematoxylin-eosin(HE) staining,silver stain, and immunohistochemical staining.The rats treated with prednisone acetate were taken as controls.Results: Observation under the light microscope with HE staining showed a sleeve-like change in rats' cerebrospinal parenchyma with inflammatory cell infiltration around the small vessels and neuronic denaturation,while silver staining showed excessive tumefaction and abscission of axon,and immunohistochemical analysis showed decreasing of nerve growth factor(NGF) expression at the acute stage of EAE,which was even more remarkable at the remission stage,showing significant difference as compared with the normal control(P0.05). And the expressions of Nogo A,an axon growth inhibitor,and its receptor(Nogo-66 receptor,Ng R) were significantly higher than those in the normal control at the acute stage(P0.01).However,after the intervention of ZGP and YGP,the pathological changes and axon damage in rats' brain and spinal cord were much more alleviated,and the NGF expression was significantly higher than that in the model group at the acute stage (P0.05).The expression of NGF was even stronger during the remission stage,and a better effect was shown by YGP.As for Nogo A and Ng R expressions,they were significantly lower than those in the model group at the acute stage(P0.05),but a better effect was shown by ZGP.Conclusions:ZGP and YGP can prevent axonal injury and promote the axonal regeneration in rats of EAE,and the possible mechanism is to increase the expression of NGF and reduce the expression of Nogo A and its receptor.However,some differences are observed between the two Chinese preparations in their acting times and points,which provides a certain basis for revealing the modern connotation of the Chinese medicine theory on tonifying Shen(肾)-yin and Shen-yang.     

Effects of Acupuncture on Monoamine Neurotransmitters in Raphe Nuclei in Obese Rats

Effects of acupuncture on the levels of neurotransmitters in the raphe nuclei were investigated in obeserats.It was found that the levels of tryptophan (Trp) and 5-hydroxyindoleacetic acid (5-HIAA) wereincreased,and 5-hydroxytryptamine (5-HT) level and 5-HT/5-HIAA ratio decreased in the raphe nucleiof the obese group as compared with the normal group;and that acupuncture could produce weightreduction,increase the 5-HT level and 5-HT/5-HIAA ratio,and decrease the contents of Trp and5-HIAA,but did not change the levels of dopamine (DA) and noradrenaline (NA).It is indicated thatbenign regulative action of acupuncture on 5-HT and its metabolism in the raphe nuclei is possibly oneof the factors for reducing weight by acupuncture.     

Effect of Stimulation of Shenshu Point on the Aging Process of Genital System in Aged Female Rats and the Role of Monoamine Neurotransmitters

In this experiment, among some aged female rats aged over 18 months, and young female rats aged 3 months whose central noradrenergic nerve endings were injured by ventricular injection of 6-hydroxy-dopamine (6-OHDA), it was observed that catgut embedding at bilateral Shenshu (UB 23) points could obviously shorten sexual cycles, increase the frequency of sexual cycle, and slow down the aging process of the genital system in both the aged rats and in the rats with injured noradrenergic endings. After electroacupuncture (EA) at Shenshu (UB 23) points in the aged rats, the frequency of neuronal discharges in locus coeruleus (LC) was elevated and the activating rate of LC to neurons in the medial preoptic area (MPOA) of the hypothalamus was increased, while obvious effect on nucleus raphes magnus (NRM) and the effect of NRM on MPOA were not marked. It is suggested that stimulation of Shenshu (UB 23) point can strengthen the excitability of noradrenergic neurons, activate the ascending pathway of the brain stem--hypothalamus, raise the catecholamine (CA)/5-hydroxytryptamine (5-HT) ratio in the hypothalamus of the aged rats, so as to delay the aging process of the genital system.     

Effects of Zuogui Pill(左归丸) and Yougui Pill(右归丸) on the Expression of Brain-Derived Neurotrophic Factor and Cyclic Adenosine Monophosphate/Protein Kinase A Signaling Transduction Pathways of Axonal Regeneration in Model Rats with Experimental Autoimm

Objective:To study the effects of Zuogui Pill(左归丸,ZGP)and Yougui Pill(右归丸,YGP)on the expressions of brain-derived neurotrophic factor(BDNF)and cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis(EAE),in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods:The rats were randomly divided into normal control(NC),model(MO),prednisone acetate(PA),ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein(MBP)_(68-86).The brain and spinal cord were harvested on the 14th and 28th day postimmunization(PI),the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction(PCR),and the cAMP levels were detected by using enzyme-immunoassay method.Results:(1)On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group(P0.05 or P0.01).(2)On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group(P0.05 or P0.01),and the cAMP level in YGP group was higher than that in ZGP group(P0.05).The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively(P0.01).(3)On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively(P0.05).(4)On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions:The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.     

Effect of Lirukang Oral Liquid(利乳康口服液) on Neurotransmitters and Estrogen in Rats with Hyperplasia of Mammary Gland

Objective: To investigate the effects of Lirukang oral liquid (LRK, 利乳康口服液) on release of neurotransmitter in rats with hyperpiasia of mammary glands (HMG) and to explore its mechanism. Methods: Sixty rats were divided into six groups, the normal control group, the model control group, the large dosage (3.6g/kg) and the small dosage (1.8g/kg) LRK groups, the Ruzengning (乳增宁, RZN, 2.5g/kg) group and the tamoxifen (TAM, 5mg/kg) group, 10 in each group. Except those in the normal control group, all the animals were made into rat model of HMG by intraperitoneal injection of estradiol benzoate. Levels of dopamine (DA) and 5-hydroxytryptamine (5-HT) in hypothalamus and mammary gland in rats were detected by fluorescence luminosity assay, and level of prolactin (PRL) in serum was detected by radioimmunoassay.Results: In the model group, the level of DA reduced significantly (P＜0.01), and 5-HT and PRL increased obviously ( P＜0.01). Compared with the model group, the LRK groups of both dosages and the TAM group had their level of DA significantly increased (P ＜ 0. 01 ), and level of 5-HT significantly decreased ( P＜0.01). The serum PRL in both LRK groups was significantly decreased ( P＜0.01). No obvious changes in DA, 5-HT and PRL were found in the RZN group. Conclusion: LRK and TAM have similar effects in regulating the release of neurotransmitter in hypothalamus and mammary gland and serum content of estrogen in the animal models of HMG.     

Effect of Jiaotai Pill (交泰丸) on Intestinal Damage in Partially Sleep Deprived Rats

Objective: To explore the effect and mechanism of Jiaotai Pill(交泰丸, JTW) on intestinal mucosal damage in rats with chronic partial sleep deprivation(PSD). Methods: Obesity resistant(OR) rats were selected, and underwent 4 h PSD by being exposed to environmental noise for 4 weeks. During the whole PSD period, JTW and estazolam were orally given to the rats respectively in the treating groups. Plasma concentration of lipopolysaccharide(LPS) which is the marker of gut-origin endotoxemia was examined. Intestinal morphology changes were observed by optical microscopy. The protein expression of occludin(Ocln) in the intestine was measured by immunofluorescence technique and Western blot. The expressions of circadian proteins cryptochromes(Cry1 and Cry2) in the intestine were also determined. Results: The treatment of JTW significantly decreased LPS level in OR rats with PSD(P0.05). JTW also attenuated insomnia-induced intestinal injury like shorter, sparse and incomplete villus, wide gap between the villus, mucosal swelling and congesting(P0.05). These changes were associated with the effect of JTW on up-regulating the expressions of Cry1 protein, Cry2 protein and Ocln protein in the intestine. Conclusions: JTW has the beneficial effect on improving intestinal mucosal damage caused by PSD. The mechanism appears to be related to the modulation of the expressions of circadian proteins and Ocln protein in the intestine, thereby attenuating inflammation and improving insulin resistance in insomnia rats.     

Jiaotai Pill (交泰丸) Alleviates Insomnia through Regulating Monoamine and Organic Cation Transporters in Rats

Objective To reveal the effect and mechanism of Jiaotai Pill(交泰丸,JTP)on insomniac rats.Methods The insomniac model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).In behavioral experiments,rats were divided into control,insomniac model,JTP[3.3 g/(kg?d)],and diazepam[4 mg/(kg?d)]groups.The treatment effect of JTP was evaluated by weight measurement(increasement of body weight),open field test(number of crossings)and forced swimming test(immobility time).A high performance liquid chromatography-electrochemical detection(HPLC-ECD)method was built to determine the concentration of monoamine transmitters in hypothalamus and peripheral organs from normal,model,JTP,citalopram[30 mg/(kg?d)],maprotiline[40 mg/(kg?d)]and bupropion[40 mg/(kg?d)]groups.Expressions of serotonin transporter(SERT),dopamine transporter(DAT),and norepinephrine transporter(NET)were analyzed by quantitative polymerase chain reaction(qPCR)and Western blot in normal,model and JTP groups.A high performance liquid chromatography-electrospray ionization mass spectrometry(HPLC-ESI-MS/MS)method was established to determine the pharmacokinetics,urine cumulative excretion of metformin in vivo,and tissue slice uptake in vitro,which were applied to assess the activity of organic cation transporters(OCTs)in hypothalamus and peripheral organs.Results Compared with the insomniac model group,the body weight and spontaneous locomotor were increased,and the immobility time was decreased after treatment with JTP(P<0.01).Both serotonin and dopamine contents in hypothalamus and peripheral organs were increased(P<0.01).The norepinephrine content was increased in peripheral organs and decreased in hypothalamus(P<0.05 or P<0.01).At the same time,SERT,DAT,OCT1,OCT2,and OCT3 were down-regulated in hypothalamus and peripheral organs(P<0.05).NET was down-regulated in peripheral organs and up-regulated in hypothalamus(P<0.05 or P<0.01).Moreover,the activity of OCTs in hypothalamus and peripheral organs was inhibited(P<0.05).Conclusion JTP alleviates insomnia through regulation of monoaminergic system and OCTs in hypothalamus and peripheral organs.     

Effects of Suxiao Jiuxin Pill (速效救心丸) on Oxidative Stress and Inflammatory Response in Rats with Experimental Atherosclerosis

Objective: To observe the preventive role of Suxiao Jiuxin Pill (SX速效救心丸) on atherosclerosis (AS) and to probe into the mechanism in the atherosclerosis rat model. Methods: The AS rat model was established by a high fat diet and a large dose of calcium (vitamin D3, 0.6 million U/kg, i.p, once). Sixty healthy male adult Sprague-Dawlay (SD) rats were randomly divided into 6 groups, a normal control group (N), a model group (M), a SX low dose group (SXL), a SX middle dose group (SXM), a SX high dose group (SXH...     

Effect of Compound Recipe Gengniankang (更年康) on Senile Sexual Hormone and Expression of Estrogen Receptor in Bone of Climacteric Female Rats

Estrogenis a vital hormone to women's wholelife.It is aninevitable process that thelevel of ser-umestrogen decreases when women enter the agedphase,and that the ovary function is exhausted.Accompanying this process,theincidence of osteo-porosis is increased and affects the quality of lifeseverely.Using hormone replacement treat ment toprevent and treat osteoporosis is very effective.Itis proved that osteoporosis is related to the lack ofestrogen,which plays a veryi mportant role duringthe proc…     

Effect of Erzhi Pill (二至丸) on Improving Cerebral Nerve Cell Apoptosis in Aging Rats

Objective: To investigate the effects of Erzhi Pill (二至丸,,EZP) on nerve cell apoptosis iinn senescence model rats.Methods: The rats model of senescence was established by peritoneal D-galactose injection combined with thymusectomy.Forty SD rats were randomized into four groups,the normal control group,the senescence model group,the EZP treated group,and the vitamins treated group,10 in each group.The rats were made into senescence model except those in the normal group.In the same time of D-galactose injection,the rats were treated respectively with distilled water,EZP 4.32 g/kg,and vitamins E and C 0.06 g/kg daily for 6 weeks via intragastric infusion.The index of main viscera (as brain,testis,etc.),serum levels of superoxide dismutase (SOD) activity,and total anti-oxidation capacity (T-AOC) were measured after a 6-week treatment.Meanwhile,the cerebral cortex neuronal apoptosis proportion and mitochondrial membrane potential (MMP) were detected by flow cytometry.Results: Both EZP and vitamins E and C treatments showed effects on increasing testis index and serum level of T-AOC,reducing the percentage of neuronal apoptosis in the cerebral cortex,and elevating MMP in the aging rats model.Conclusions: EZP could inhibit the cerebral cortex neuron apoptosis and maintain the mitochondrial function in the senescent process of rats induced by peritoneal D-galactose injection combined with thymusectomy.It also shows antioxidation effect to some extents.     

Effect of Acute Pancreatitis on the Pharmacokinetics of Chinese Herbal Micron Liuhe Pill Ointment (微米六合丹) in Rats

Objective: To explore the effect of acute pancreatitis (AP) on the pharmacokinetics of herbal ointment micron Liuhe Pill (微米六合丹, MLHP) components in anesthetized rats. Methods: Rats were randomly divided into a AP model group (n=6) and a normal group as a control (n=6). The rat model of AP was induced by intraperitoneal injection of L-arginine in rats (15 mg/kg, twice, interval 1 h). Chinese herbal ointment MLHP was used externally on the belly after the 2nd injection for 48 h in both groups. Emodin, rhein, aloe emodin, physcion, chrysophanol from MLHP were detected and quantified in rat serum and pancreas (at 48 h) by high performance liquid chromatography-tandem mass spectrometry. Results: Among the five components, only emodin, aloe emodin and physcion from MLHP were detected in all rat serum and most of the rats'' pancreas. Rhein and chrysophanol were not detected in both serum and pancreas. T1/2α of emodin and physcion in MLHP were obviously shorter in the AP model group than those in the normal group (P<0.05), while there was no difference for T1/2α of aloe emodin. The peak concentration and area under curve of all three components were much higher in the AP group than those in the normal group with MLHP in external application for 48 h (P<0.05). Furthermore, the mean residence time (MRT) and maximum plasma concentration (Tmax) of emodin and aloe emodin were obviously longer in the AP model group than those in the normal control group (P<0.05). There was no significant difference for Ka of all components between the two groups. Emodin could be detected in all rats'' pancreas at 48 h in both groups, while its mean pancreatic concentration was higher in the AP model group than in the normal group (0.61±0.54 ng/mL, 0.42±0.37 ng/mL, respectively, P<0.05). Aloe emodin could be detected in all rats'' pancreas at 48 h in both groups and their mean pancreatic concentration were similar (0.31±0.24 ng/mL, 0.33±0.17 ng/mL, respectively, P>0.05). Physcion could be detected in pancreas of most rats in the AP model while only two rats in the normal group. Conclusion: AP could significantly affect the pharmacokinetics of absorbed components of Chinese herbal MLHP ointment in rats.     

Brain Monoamine Contents and Antiulcerogenic Effects of Furazolidone in Rats

Our previous work showed that, in the rats withreserpine-induced gastric ulcer, brain monamines (NE, DAand 5-HT) were exhausted and furazolidone completely pre-vented such a gastric ulcer. The present study was carriedout to observe brain monoamine contents andantiuloerogenic effects of furazolidone in other gastric ulcermodels of rats. It was found that in the rats with gastric ul-oer induced by restraint in water (21℃ for 18 h), brainNE content was significantly decreased. Furazolidone partial-ly prevented this gastric ulcer and increased brain     

Effect of the Recipe of Zhishi Xiaopi Pill and its Ingredients on Gastric Emptying Rate and Plasma Motilin Level in Rats

To investigate the influence of the recipe of Zhishi Xiaopi Pill （ZSXPP） and its ingredients on gastric emptying and plasma motilin （MOT） level in rats. After the rats were douched by different herbs, cisapride or normal saline in different groups for 4 weeks, The gastric emptying rate and plasma MOT levels were measured respectively. The gastric emptying rate and plasma motilin levels were both increased significantly by using the total recipe of ZSXPP and some parts of the recipe. ZSXPP can increase the gastric emptying rate and plasma motilin levels in rats. The herbs can remove food and qi stagnancy and relieve flatulence; Zhishi, Houpu and Maiya were the main effective ingredients in this recipe.     

Determination and Comparison of Monoamine Neurotransmitters in Rat Striatum by High Performance Liquid Chromatography with Fluorescence Detection

A method for separation and quantitative determination of,monoamine transmitters （norepinephrine, dopamine, dihydroxyphenylalanine, 5-hydroxytryptamine） in rat striatum by high performance liquid chromatography with fluorescence detection analysis was described. Under optimized chromatographic conditions, four monoamine transmitters were successfully separated and detected. This method showed high sensitivity and resolution. In addition we established the model of Parldnson＇s Disease rat and impaired its ventral pallium observe changes of the rotational behavior and the content of monoamine transmitters in striatum. In conclusion, this method is highly suitable for studying changes in monoamine transmitter＇s levels in neuropsychiatric disorders. The detection limits （signal-to-noise ratio=3） for monoamine transmitters were 1.0 pmoL-3.0 pmoL. The relative standard deviation of the peak-heights ratio are about 1.9%, 2,2%, 0.7%, and 2.6% for norepinephrine, dopamine, dihydroxyphenylalanine, and 5-hydroxytryptamine, respectively （n=5）. It exhibits the expected linear behavior over the quantity range from 2 pmoL to 300 pmoL and the correlation coefficients are all greater than 0.999. The rotational behavior of Parkinson＇s Disease rats was significantly improved by ventral pallidum lesion and the content of monoamine transmitters also changed remarkably.     

Effect of Modified Wuzi Yanzong Pill(五子衍宗丸) on Tip60-Mediated Apoptosis in Testis of Male Rats after Microwave Radiation

Objective: To investigate the effect of a modi?ed Wuzi Yanzong Pill(五子衍宗丸, WZYZP) on the male rats' testis after microwave radiation, as well as its potential mechanism. Methods: Forty-?ve male rats were randomly assigned to three groups: the control group, the radiation group, and the WZYZP group. The rats in the radiation group and WZYZP group were exposed to microwave radiation for 15 min once, while the rats in the control group were not exposed to any radiation. The rats in the WZYZP group were given a modi?ed of WZYZP by gavage daily for 7 days. Apoptosis in the testis was evaluated using terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) assay. Histopathological alterations of the testis were observed by haematoxylin-eosin(HE) staining. Tat-interactive protein, 60 kD(Tip60) and p53 expressions were determined by Western blotting. Results: The apoptosis index(AI) in the radiation group was higher than that of the WZYZP group and control group on day 1(D1), day 7(D7) day 14(D14) after radiation(P0.05). The seminiferous tubules were of normal morphology in the control group. In the radiation group, the partial seminiferous tubules were collapsed, basement membranes of the seminiferous epithelia became detached. WZYZP could restore the morphological changes. There was no expression of Tip60 among the three groups on D7 and D14. The expression of p53 was higher in the radiation group than in the control group(P0.05). WZYZP could down-regulate the rising p53 induced by radiation on D7 and D14(P0.05). Conclusion: A modified WZYZP may affect germ cells, and its protective effects may partly result from its ability to intervene in Tip60 mediated apoptosis.     

Effect of Moniliformin on Myocardial Contractility in Rats

The present study was carried out on rat heart to in situ determine the myocardial toxicity of moniliformin, a synthesized compound, originally isolated from mouldy corn and soil samples in the Keshan disease prevalent area in China. Perfusion of moniliformin 10-7 mol/liter in isolated heart decreased myocardial contractile force by 52±17%. Intravenous injection of moniliformin at 1/6 and 1/4 LD_(50) markedly inhibited cardiac hemodynamic variables relative to myocardial contractile function. Particularly, it decreased 4±LV dP/dt max by 52±6%, and induced ventricular arrhythmia. These data indicate that moniliformin is toxic to mammalian heart and might be an important factor relative to Keshan disease.     

Effect of Formaldehyde on Spermatogenesis and Testicular Morphology in Adult Rats

Objective To study the effects of formaldehyde （FA） on spermatogenesis and testicular morphology in adult rats. Methods The rats were divided at random into two experimental groups and one control group, each comprising 10 individuals. Rats in two experimental groups were respectively exposed to FA at concentrations of 0.5mg.m^-3 and 10mg.m^-3 （12h per day） for 2 weeks. After the treatment, the testicular weight and morphology, as well as the quantity and quality of sperm were evaluated. Results compared with the control, the testicular weight, sperm count, the percentage of motile sperm significantly decreased （p〈0.05）, while the percent of abnormal sperm significantly increased （p〈0.05） in rats of 10mg.m^-3 FA exposure group. Microscopy with HE staining showed that seminiferous tubules atrophy, spermatogenic cells decreased, seminiferous epithelial cells disintegrated and shed in rats of 10mg.m^-3 FA exposure group. Conclusion FA has a harmful effect on the spermatogenesis; it induces tubular atrophy and sperm abnormality at a dose-dependent manner.     

Effect of Ginseng Injection in Treating Congestive Heart Failure and Its Effect on Thyroid Hormones

ONective：ToobservetheeffectofGinsenginjectiononcongestiveheartfailure（CHF）anditseffectonthyroidhormones．Methods：Patients（n＝54）ofCHFweredividedintotwogroups．Onthebasisofconventionaltreatmentofwesternmedicine，theGinsenginjectionwasgiventothetreatedgroupadditionallyfortwoweeks．Bloodleveloftriiodothyronine（T3），thyroxine（T4），reversetriiodohyronine（rT3）andthyroidstimulatinghormone（TSH）ofallpatientsweredeterminedbeforeandaftertreatmentbyradioimmunoassay．Results：Therewas…