Objective: To establish two methods (HPLC UV and HPLC ELSD)for the determination of 3 kinds of cholic acids in Qingkailing Injection, and compare them with UV method. Methods: Two HPLC methods detected by UV and ELSD, respectively have been developed: Phenomenex Luna C 18 column(250mm?4.6mm,5?)with linear gradient elution using methanol acetotrile(4∶1) and 0.1% aqueous formic acid. Results: Three kinds of cholic acids in Qingkailing could be determined simultaneously by HPLC UV and HPLC ELSD. The recovery of three method were 100%,97%,95%～96% for HPLC/ELSD,HPLC/UV,UV, respectively. Conclusion: The HPLC ELSD method was accurate and specific to determine the content of cholic acids in Qingkailing Injection.

Objective To analyze the relationship between cytotoxic T cells Perforin level, IFN-γand IL-10 in patients with chronic hepatitis B. Methods After a short-term cultivation of peripheral blood collected from 50 patients with chronic hepatitis B, a flow cytometry was employed to detect the levels of Per-forin, IFN-γ and IL-10 in CD8~+ cells to compare with those of the health donors the HBV DNA copies in their blood were also measured by RT-PCR to analyze the relationship with Perforin, IFN-γ, and IL-10 in CD8~+ cells. Results In patients with chronic hepatitis B, the levels of Perforin, IFN-γ and IL-10 in CD8~+ ceils were (5.30 ± 2.62)%, (4.05 ± 2.25) % and (0.77 ± 0.50) %, respectively, which were statistical-ly lower than health donors(the t values were 4.50, 4.56 and 4.20 respectively; P < 0.01) ; Of 26 cases of chronic hepatitis B patients with HBeAg-positive Pefforin and IFN-γ in CD8~+ T cells were (4.54 ± 1.93) % and (3.32 ± 1.59)%, respectively, significantly lower than those of the 24 chronic hepatitis B patients with HBeAg-negative (the t values were 2.22 and 2.54, respectively; P <0.05) ; And the levels of Perforin, IFN-γ had a negative relation with the HBV DNA copies (the coefficient correlations-0. 539 and-0. 340; P < 0.01 and P < 0.05). Conclusion In patients with chronic hepatitis B, the reduced levels of Pefforin, IFN-γ and IL-10 may be related with the long-term existence of HBV, and the protracted course of disease.

AIM: To study the determination of 5 kinds of cholic acid content. METHODS: Using single standard substitue (SSS), we assayed 5 kinds of cholic acid, such as cholic acid, deoxycholic acid, hyodexycholic acid, chenodeoxychoilc acid and ursodeoxycholic acid by HPLC/ELSD. and studied the influence of the additive substitutes (such as rabbit Serum, urine and Qingkailing extract) on the measure values. RESULTS: By using any one of the 5 kinds of cholic acid standard, the content in all the 5 kinds of cholic acid could be determined. CONCLUSION: Simply as well as credibly, the SSS method could be used in quality analysis, especially in complex matrix. Also, it proved a feasible way in quality control and research of fingerprint chromatography of TCM.

Objective To explore the effects of prostaglandin E1 (PGE1) on the expression of Apaf-1 and TLR4 in rats with cerebral ischemia-reperfusion(CIR) injury.Methods 32 healthy adult male Wistar rats were randomly divided into four groups,which were sham operated group (n=8),CIR model group (n=8) and PGE1 pretreated groups (low dose,12 μg · kg-1;high dose,24 μg · kg-1,n =8).Rat model of cerebral ischemia/reperfusion was established by bilateral common carotid artery ligation.The expression of Apaf-1 and TLR4 was detected with immunohistochemical staining method in hippocampus and epencephalon.Results After 20 min of ischemia and reperfusion for 24 h,compared with sham operated group (Apaf-1:hippocampus (0.87±0.78),epencephalon (0.67 ±0.43);TLR4:hippocampus (2.43 ± 1.17),epencephalon (1.97± 1.033)),the number of positive cells of Apaf-1 (hippocampus (11.83± 2.26);epencephalon(5.80±1.30) and TLR4 (hippocampus(16.90±2.86);epencephalon(12.90±2.66)) was increased in CIR model group (P＜0.05).Compared with CIR model group,the positive cell numbers of Apaf-1 (hippocampus:low dose(9.83±2.12),high dose(5.50± 1.17);epencephalon:low dose(4.87± 1.38),high dose(2.73±1.172)) and TLR4 (hippocampus:low dose (11.53± 2.40),high dose (9.13 ± 2.54);epencephalon:low dose (9.07 ± 2.07),high dose (4.47 ± 1.68)) were reduced dose-dependently in PGE 1 pretreatment all group (all P ＜0.05).Conclusion PGE1 can inhibit the expression of Apaf-1 and TLR4 in hippocampus and epencephalon of rat with cerebral ischemia-reperfusion injury.

Objective To observe and compare in vivo segmental lumbar motion between symptomatic L4 isthmic spondy?lolisthesis (IS) patients and L4 degenerative spondylolisthesis (DS) patients during functional weight?bearing activities. Methods Fifteen symptomatic L4 IS patients and fifteen symptomatic L4 DS patients were recruited. Fifteen asymptomatic volunteers were en?rolled as the control group. The L4,5 vertebral segment motion of each subject was reconstructed using three?dimensional computed tomography and a solid modeling software. In vivo, lumbar vertebral motion during functional postures (flexion?extension, left?right twisting and left?right bending) was observed using a dual fluoroscopic imaging technique. The spinal function unit was divided in?to anterior and posterior segments by the isthmic cleft. Local coordinate systems were established at the vertebral body, to obtain the 6 degree?of?freedom (DOF) intervertebral range of motion (ROM) at L4,5 and the ROM between the anterior and the posterior segments of L4 IS. Results The motion of IS at L4,5 was found to be increased. The migrations along both sagittal and vertical axis were significantly larger than control group (P<0.05). During left?right twisting, the migration along sagittal axis was significantly larger than control group (P<0.05);the rotation along vertical axis was significantly larger than DS and control group (P<0.05). During left?right bending, the migration along frontal axis was significantly larger than DS and control group ( P<0.05). The inter?vertebral ROM at L4,5 showed no significant difference between DS and control group. The migration between anterior and posteri?or segments of IS L4 was larger in standing than supine (P<0.05). The relative migration along sagittal axis between anterior and posterior segments of IS L4 was significantly larger in flexion than standing (P<0.05). Conclusion A spondylolytic defect does lead to detectable instability or hypermobility in the lumbar spine compared with degenerative spondylolisthesis and normal group. From supine to standing posture, migration at isthmic cleft along sagittal and coronal axis are identified in IS spondylolithesis.

Objective The objective of this study was to investigate the expression of P28 and P43 in papillary thyroid carcinoma(PTC)and its relationship with clinicopathological features after TRa1 gene transcription.Methods Real-time fluorescence quantitative PCR(RT-PCR)and gel electrophoresis were used to determine the target fragments and to isolate the bands of different fragments.The optical density of each band was scanned by UV transmittance analyzer to detect 31 cases of PTC tissue and expression levels of P28 and P43 in para-cancerous tissues.Results The average gray value of P28 in thyroid carcinoma group was 0.77±0.34,which was significantly higher than that in para-cancer group(0.31±0.18).The average gray value of P43(0.85+0.21)in thyroid carcinoma group was significantly higher than that in para-cancer group(0.34±0.15)(P<0.05).The expression levels of P28 were not correlated with gender,age,tumor size,lymph node metastasis and clinical pathologic stage(P>0.05),The expression levels of P43 were not correlated with gender,age,tumor size and lymph node metastasis.(P>0.05)but they were related to clinical pathologic stage(P<0.05).There was no correlation between expression levels of P28 and P43(r=0.266,P=0.071).Conclusion The increased expression of P28 and P43 may have a high degree of malignancy and a certain clinical value in predicting the adverse prognosis of PTC.Both factors are helpful for the prevention and treatment of PTC.

Objective:This study aimed to achieve the early diagnosis and active treatment of adult hemophagocytic syndrome (HPS) and investigate the clinical characteristics and prognostic factors of this syndrome. Methods:A single-center retrospective analysis was performed to analyze clinical characteristics, laboratory findings, and survival data. Results:In 58 patients, the most common clinical manifestations were fever (100%) and splenomegaly (89.7%). The most common laboratory parameters were serum ferritin 500 g/L (100%) and peripheral cytopenia in two or more lineages (96.6%). platelet count, fibrinogen, and lactate dehydrogenase in the death group were significantly lower than in the survival group (P=0.000, 0.001, and 0.000). Survival analysis results showed that infections in the rheu-matological group exhibited good prognosis [the overall survival (OS) time was not reached in 190 d]. Patients with unexplained causes had moderate prognosis (OS time was 60 d);tumor-associated HPS patients had poor prognosis (the OS time was only 30 d). Univariate analysis results showed that patients with Fbg<1.5 g/L, PLT<40×109/L, and LDH≥2000 U/L also exhibited poor prognosis (P=0.000). Multivariate analysis results showed that PLT<40 × 109/L was an independent adverse factor (HR=6.472, 95%CI:1.526-26.065, P=0.011). Conclusion:HPS exhibits complex clinical manifestations and varied etiology. Patients with infection and rheumatism-related HPS had good prognosiss compared with those manifesting tumor-associated HPS. Fbg<1.5 g/L, PLT<40×109/L, and LDH≥2 000 U/L were the univariate factors that affected the survival time of patients. PLT<40×109/L is an independent adverse factor. These patients need systemic treatments as early as possible.

Objective To clarify the expression of leptin and leptin receptor in normal brain tissues and gliomas and investigate the effect of exogenous leptin on the proliferation,migration and invasion of human glioma U251 cell line.Methods Immunohistochemical staining was used to detect the expression of leptin and leptin receptor in 50 cases of different grades of glioma tissues and 20 cases of normal brain tissues.The effects of exogenous leptin on proliferation,migration and invasion of U251 cells were detected by MTT assay,cell scratch assay and Transwell invasion assay.Results (1) The positive expression rates of leptin and leptin receptors in glioma tissues were 50.0％ and 92.0％,respectively.(2)Proliferation activity:leptin concentrations of 0 ng/ml,10 ng/ml,and 50 ng/ml had no significant difference in the proliferation of U251 cells (absorbance:0.263±0.015,0.273±0.017 and 0.277±0.006,respectively),and the leptin concentration of 100 ng/ml had a significant effect on the proliferation of U251 cells (absorbance:0.315±0.005,P＜0.05).(3)Migration ability:the migration rate of U251 cells treated with different concentrations of leptin increased significantly with the passage of time,and the migration rate was most significant at the concentration of 100 ng/ml ((93.313±3.080) ％),and the difference was statistically significant (P＜0.05).(4)Invasive ability:with the increase of leptin concentration and the prolongation of the action time,the invasive ability of U251 cells was enhanced.When leptin was used at a concentration of 100 ng/ml,the number of penetrating cells were the biggest(135±2).Conclusion Leptin and leptin receptors are involved in the occurrence of gliomas;and exogenous leptin promotes the proliferation of U251 cells and has time and dose dependability on the migration and invasion of U251 cells.

Objective To evaluate the diagnostic value of acetic acid spray combined with narrow-band imaging ( NBI) for early gastric cancer and precancerous lesion. Methods A total of 136 gastric cancers and precancerous lesions from 132 patients detected by screening endoscopy from November 2015 to November 2017 in Guangming Chinese Medicine Hospital of Pudong New Area, Shanghai were enrolled in this study, and were divided into NBI group, acetic acid spray group, and acetic acid spray combined with NBI group ( combination group) . The image clarity, microsurface pattern, microvascular pattern, demarcation line, and mucosal whitening time were observed, and value of the 3 methods in diagnosis of early gastric cancer and precancerous lesions was analyzed. Results Image clarity of micro glandular in combination group was significantly better than that in NBI group and acetic acid spray group (χ2=8. 766, P=0. 003;χ2=5. 273, P=0. 022) , and image clarity of microvascular in combination group was significantly better than that in NBI group (χ2=7. 457, P=0. 006) . The overall diagnostic coincidence rate with pathology of combination group, NBI group, and acetic acid spray group was 91. 9％(125/136), 85. 3％(116/136), and 89. 7％(122/136), respectively. The diagnostic sensitivity of irregular or missing microsurface, irregular or missing microvascular, obvious demarcation line, mucosal whitening time <30 s for carcinoma ( including high grade intraepithelial neoplasia, early carcinoma and infiltrating carcinoma ) were 92. 3％, 91. 3％, 92. 3％ and 90. 4％, respectively, in the combination group, and the diagnostic specificity of above indicators were 93. 8％, 93. 8％, 96. 9％ and 90. 6％, respectively, accuracy were 92. 6％, 91. 9％, 93. 4％ and 90. 4％, respectively. For observation of microsurface, the combination group was superior to the NBI group (χ2 =7. 378, P=0. 007) , but there was no significant difference compared with the acetic acid spray group (χ2=0. 427, P=0. 513);the acetic acid spray group was superior to the NBI group (χ2=4. 405, P=0. 036) . For observation of microvascular, the combination group was not significantly better than the NBI group (χ2=2. 398, P=0. 122). For observation of demarcation line, the combination group was not significantly better than the NBI group (χ2=2. 722,P=0. 099) and the acetic acid spray group (χ2=0. 216, P=0. 642). There was no difference between the acetic acid spray group and the NBI group (χ2=1. 433, P=0. 231) in observation of demarcation line. Conclusion Acetic acid spray combined with NBI has a good consistency with pathological results with high diagnostic sensitivity and specificity, and can clearly show microsurface of early gastric cancers and precancerous lesions compared to single NBI.

ObjectiveTo observe the clinical efficacy of tonifying kidney and replenishing essence on asthenozoospermia patients with the syndrome of kidney essence deficiency and the effects of this method on the adenosine 5′-monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin complex 1 （mTORC1） signaling pathway. MethodsSeventy-two eligible asthenozoospermia patients with the syndrome of kidney essence deficiency treated in the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from February 2023 to January 2024 were selected and randomly assigned into an observation group and a control group， with 36 patients in each group. The observation group received oral administration of Guilu Tianjing capsules， while the control group received oral administration of L-carnitine oral solution. The treatment course lasted for 4 weeks in both groups. The observed indicators included sperm progressive motility rate （PR）， total sperm motility （PR+NP）， percentage of normal mitochondrial membrane potential （MMP）， and traditional Chinese medicine （TCM） symptom scores before and after treatment in both groups. A three-month follow-up was instituted to record the conception status of the patients’ spouses. Additionally， eight patients were randomly selected from the eligible patients in the observation group， and four healthy males with normal semen routine examination results were included as the control group for the determination of protein expression. Western blotting was conducted to assess the expression of AMPK， phosphorylated （p）-AMPK， regulatory-associated protein of mTOR （RAPTOR） and p-RAPTOR， and PTEN-induced putative kinase 1 （PINK1） in sperms from the observation group before and after treatment， as well as in the sperms of the control group. ResultsThe pregnancy rate of spouses in the observation group was 9.09% （3/33）， which was higher than that （3.33%， 1/30） in the control group. The total response rate was 84.8% （28/33） in the observation group and 66.7% （20/30） in the control group， with no statistically significant difference. After treatment， both groups were improved considering PR， PR+NP， MMP， and TCM symptom scores （P<0.01）. Moreover， the observation group exhibited more pronounced decreases in TCM symptom scores than the control group （P<0.05）， while the changes in PR， PR+NP， and MMP showed no statistical significance between groups. Compared with the control group， the asthenozoospermia group exhibited upregulations in phosphorylation levels of AMPK and RAPTOR and protein level of PINK （P<0.01）. The administration of Guilu Tianjing Capsules led to downregulations in the phosphorylation levels of AMPK and RAPTOR and protein level of PINK1 （P<0.01）. However， the protein levels of AMPK and RAPTOR demonstrated no significant difference between before and after treatment. During the study period， neither group of patients exhibited any notable adverse reactions. ConclusionGuilu Tianjing capsules can enhance the sperm motility and percentage of normal mitochondrial membrane potential in asthenozoospermia patients with the syndrome of kidney essence deficiency by downregulating the AMPK/mTORC1 signaling pathway， lowering the protein level of PINK1， and inhibiting excessive activation of mitophagy.