Objective To explore the mechanism by which paeoniflorin improves trabecular meshwork(TM)cell dysfunction.Methods C57BL/6J mice were randomly divided into the following groups:Control group(no treatment),model group(glaucoma model constructed by ocular perfusion)and paeoniflorin group(glaucoma model followed by treatment with 50 mg·kg-1 paeoniflorin).Each group consisted of 12 mice.After 14 days of treatment,intraocular pressure was measured;and trabecular meshwork tissue was collected.TdT mediated dUDP nick end labeling(TUNEL)assay was performed to detect cell apoptosis.Western blot was used to measure protein expression levels.HTMC cells were randomly divided into the following groups:Normal group(cultured under standard conditions),rapamycin group(50 μmol·L-1 rapamycin as an autophagy inducer),experimental-L group(50 μmol·L-1 rapamycin+10 μmol·L-1 paeoniflorin)and experimental-H group(50 μmol·L-1 rapamycin+30μmol·L-1 paeoniflorin).Western blot was used to measure protein expression levels.Methyl thiazolyl tetrazolium(MTT)assay and flow cytometry were applied to evaluate cell proliferation and apoptosis.Results After 14 days of treatment,the intraocular pressure in the control,model and paeoniflorin groups were(12.81±0.83),(26.31±1.85)and(20.64±1.77)mmHg,respectively;the positive cell rate detected by the TUNEL assay were(4.86±0.44)％,(30.32±5.15)％and(15.08±1.92)％,respectively;the levels of microtubule associated protein 1 light chain 3(LC3)Ⅱ/LC3 Ⅰ protein were 0.51±0.06,1.33±0.13 and 0.72±0.08,respectively;Collagen Ⅰ protein levels were 0.45±0.07,1.11±0.12 and 0.72±0.05,respectively.The above indexes of the model group were statistically significant compared with the control group,and the above indexes of the paeoniflorin group were statistically significant compared with the model group(P＜0.001,P＜0.01).The LC3 Ⅱ/LC3 Ⅰ protein levels in the normal,rapamycin,experimental-L,experimental-H groups were 0.40±0.03,1.54±0.10,0.98±0.10 and 0.64±0.06,respectively;the cell viability rates were(100.00±6.25)％,(65.96±6.16)％,(75.22±5.54)％and(82.15±5.14)％,respectively;the apoptosis rates were(4.80±0.37)％,(19.64±0.97)％,(16.10±0.93)％and(13.16±0.94)％,respectively.The above indexes in the rapamycin group were significantly different from those in the normal group,and the above indexes in the experimental-L,-H groups were significantly different from those in the rapamycin group(P＜0.001,P＜0.05).Conclusion Paeoniflorin may improve trabecular meshwork cell dysfunction by regulating autophagy and reducing cell apoptosis,which may provide potential therapeutic strategies for glaucoma.

Objective To explore the mechanism by which paeoniflorin improves trabecular meshwork(TM)cell dysfunction.Methods C57BL/6J mice were randomly divided into the following groups:Control group(no treatment),model group(glaucoma model constructed by ocular perfusion)and paeoniflorin group(glaucoma model followed by treatment with 50 mg·kg-1 paeoniflorin).Each group consisted of 12 mice.After 14 days of treatment,intraocular pressure was measured;and trabecular meshwork tissue was collected.TdT mediated dUDP nick end labeling(TUNEL)assay was performed to detect cell apoptosis.Western blot was used to measure protein expression levels.HTMC cells were randomly divided into the following groups:Normal group(cultured under standard conditions),rapamycin group(50 μmol·L-1 rapamycin as an autophagy inducer),experimental-L group(50 μmol·L-1 rapamycin+10 μmol·L-1 paeoniflorin)and experimental-H group(50 μmol·L-1 rapamycin+30μmol·L-1 paeoniflorin).Western blot was used to measure protein expression levels.Methyl thiazolyl tetrazolium(MTT)assay and flow cytometry were applied to evaluate cell proliferation and apoptosis.Results After 14 days of treatment,the intraocular pressure in the control,model and paeoniflorin groups were(12.81±0.83),(26.31±1.85)and(20.64±1.77)mmHg,respectively;the positive cell rate detected by the TUNEL assay were(4.86±0.44)％,(30.32±5.15)％and(15.08±1.92)％,respectively;the levels of microtubule associated protein 1 light chain 3(LC3)Ⅱ/LC3 Ⅰ protein were 0.51±0.06,1.33±0.13 and 0.72±0.08,respectively;Collagen Ⅰ protein levels were 0.45±0.07,1.11±0.12 and 0.72±0.05,respectively.The above indexes of the model group were statistically significant compared with the control group,and the above indexes of the paeoniflorin group were statistically significant compared with the model group(P＜0.001,P＜0.01).The LC3 Ⅱ/LC3 Ⅰ protein levels in the normal,rapamycin,experimental-L,experimental-H groups were 0.40±0.03,1.54±0.10,0.98±0.10 and 0.64±0.06,respectively;the cell viability rates were(100.00±6.25)％,(65.96±6.16)％,(75.22±5.54)％and(82.15±5.14)％,respectively;the apoptosis rates were(4.80±0.37)％,(19.64±0.97)％,(16.10±0.93)％and(13.16±0.94)％,respectively.The above indexes in the rapamycin group were significantly different from those in the normal group,and the above indexes in the experimental-L,-H groups were significantly different from those in the rapamycin group(P＜0.001,P＜0.05).Conclusion Paeoniflorin may improve trabecular meshwork cell dysfunction by regulating autophagy and reducing cell apoptosis,which may provide potential therapeutic strategies for glaucoma.

Abstract： Objective　To analyze the value and influencing factors of cross-primer isothermal amplification technology(CPA) in clinical screening and diagnosis of tuberculosis (TB). Methods　We collected 543 inpatients in the Second Affiliated Hospital of Hainan Medical College from January 1, 2018 to December 31, 2021, including 179 patients with tuberculosis, 187 patients with pneumonia and 177 patients with other diseases. The patients' sputum, alveolar lavage fluid, pleural effusion and midstream urine were detected by CPA, smear microscopy, culture method and gene detection. The value of CPA detection in the diagnosis of tuberculosis and its influencing factors were evaluated. Statistical analysis was performed using SPSS 26.0. Results　The total positive rate of CPA was 14.4% (78/543), and the positive rate of sputum samples accounted for 29.1% (39/134). Among the 78 cases of CPA positive patients, the tuberculosis group accounted for 69.2% (54/78), followed by pneumonia group 21.8% (17/78), and other diseases group accounted for 9.0% (7/78). Taking CPA test as the reference method, the "sensitivity" of smear microscopy was lower than that of genetic testing and culture, while the "specificity" was higher than that of culture and gene testing, and the "missed diagnosis rate" of smear microscopy was higher than that of genetic testing and culture. CPA test positive was related to gender, ESR and pneumonia. There is a good agreement between CPA test and culture method and gene test (Kappa>0.9), and a moderate agreement between CPA test and smear microscopy (Kappa=0.616). Conclusions　Sputum specimen is the best choice for CPA detection, while the value of pleural effusion detection is relatively limited. Sputum, alveolar lavage fluid and midcourse urine can be used as clinical specimens for screening and diagnosis of "tuberculosis group and other disease group", while sputum can be used for screening and diagnosis of "tuberculosis group and pneumonia group". Gender, ESR and pneumonia are the influencing factors of CPA positive patients. Therefore, CPA testing is worthy of clinical promotion, but more clinical research data are needed.

Eleven compounds were isolated from the 95% ethanol extract of the stems of Dendrobium officinale after water extraction by various modern chromatographic techniques, such as silica gel column chromatography(CC), octadecyl-silica(ODS) CC, Sephadex LH-20 CC, preparative thin layer chromatography(PTLC) and preparative high performance liquid chromatography(PHPLC). According to spectroscopic analyses(MS, 1D-NMR, 2D-NMR) combined with optical rotation data and calculated electronic circular dichroism(ECD), their structures were identified as dendrocandin Y(1), 4,4'-dihydroxybibenzyl(2), 3-hydroxy-4',5-dimethoxybibenzyl(3), 3,3'-dihydroxy-5-methoxybibenzyl(4), 3-hydroxy-3',4',5-trimethoxybibenzyl(5), crepidatin(6), alternariol(7), 4-hydroxy-3-methoxypropiophenone(8), 3-hydroxy-4,5-dimethoxypropiophenone(9), auriculatum A(10) and hyperalcohol(11). Among them, compound 1 was a new bibenzyl derivative; compounds 2 and 7-11 have not been previously reported from Dendrobium plants; compound 6 was reported from D.officinale for the first time. Compounds 3-6 exhibited potent antioxidant activity with IC_(50) values of 3.11-9.05 μmol·L~(-1) in ABTS radical scavenging assay. Compound 4 showed significant inhibitory effect on α-glucosidase, with IC_(50) value of 17.42 μmol·L~(-1), indicating that it boasted hypoglycemic activity.
Dendrobium ; Biological Assay ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Bibenzyls

BACKGROUND:Three-dimensional (3D) printing technology is currently one of the most advanced industrial manufacturing technologies. The surgical template prepared based on the 3D printing technology is mainly made of resin, and a great improvement in its accuracy is required. However, the clinical application of the surgical template made of metal is rarely reported. OBJECTIVE:To evaluate the clinical value of 3D-printing implant template in the restoration of free-end missing teeth.METHODS:A prospective study was conducted in 64 enrolled patients with free end-tooth defects. All the patients were randomly assigned to receive traditional implant template (control group,n=32) or 3D-printing implant template (study group,n=32), and 3-6 months later, the patients were subjected to crown restoration. At 6 months after crown restoration, cone beam computed tomography was performed to compare the deviation of the implant tip and neck (including vertical, buccolingual, mesial-distal). Success rate and chewing rate were compared between the two groups at 6 months after crown restoration; patient satisfaction assessment was done and compared between the two groups at 1 year after crown restoration. RESULTS AND CONCLUSION:There were no significant differences between the two groups in the success rate and chewing rate (98.7％ vs. 95.6％; 97.4％ vs. 97.1％,P>0.05). The vertical, buccolingual, mesial-distal deviations of the implant tip were significantly lower in the study group than the control group (P<0.05), while there was no difference in the vertical and buccolingual deviations of the implant neck between the two groups (P>0.05), and the mesial-distal deviation of the implant neck was significantly lower in the study group than the control group (P<0.05). In addition, there was no difference in the patient satisfaction between the study and control groups (94％vs. 91％, P>0.05). To conclude, the 3D printing implant template can effectively reduce implant excursion based on the assurance of therapeutic efficacy and patient satisfaction, which is of great significance in the restoration of free-end tooth loss.

[Objective]To investigate the characteristics of immunophenotypes in hepatocellular carcinoma(HCC) before liver transplantation.[Methods]The immunophenotypes of T-,B- cells,monocytes,dendritic cells(DC)and NK-cells in peripheral blood from 6 HCC patient who were ready to have liver transplantation and 6 healthy volunteers were analyzed by multicolor flow cytometry.[Results]In the patients,the proportions of CD4+PD-1+T cells,Treg cell (CD4+CD25+CD39+T cells),CD19+B cells,Plasmablasts(CD27highCD38highIgD-IgM-),classical monocytes(CD14high CD16-)and mature NK-cells(CD3-CD56high)were all higher than those in the healthy controls(all P<0.05).However, marginal zone B cell(CD27+IgD+),Non-switched B cells(CD27+CD38dimIgM+),intermediate monocytes(CD14high CD16+)and immature NK-cells(CD3-CD56+)were lower than those in the healthy controls(all P<0.05). And there wasn't any obvious difference in quantity being observed among other cell types.[Conclusion]There was difference in the immunophenotypes of immune cells in peripheral blood between HCC patients before liver transplantation and healthy people.And this finding exerts important effects on monitoring the immune status of the patients after liver transplantation and guiding the administrations of immunosuppressors.

This study aimed to analyze the analgesic effect and related central mechanisms of CQ prescription on cancer invasion induced mirror image pain (CIIMIP)in model mice.In the study, male BALB/c mice were randomly divided into normal group, operation control group (injected with 0.2 mL inactivated S180 sarcoma cell sap), model group (injected with 0.2 mL S180 sarcoma cell sap on the right leg near the greater trochanter of femur) and CQ prescription low dose group (intraperitoneally injected with CQ prescription 100 mg•kg⁻¹ on the basis of model mice), CQ prescription middle dose group (intraperitoneally injected with CQ prescription 150 mg•kg⁻¹ on the basis of model mice), and CQ prescription high dose group (intraperitoneally injected with CQ prescription 200 mg•kg⁻¹ on the basis of model mice). Mechanical withdraw threshold (MWT) of the mirror image lateral hind paws were evaluated by Von Frey hairs before modeling and after surgery. The levels of glutamate (Glu), gamma aminobutyric acid (GABA), glycine (Gly), and taurine (Tau) in the L3-L5 spinal cord were measured by the high performance liquid chromatography-fluorescence detector (HPLC-FLD); AimPlex detection technology with multiple factors was used to detect the levels of regulated on activation in normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein (MCP-3) in the L3-L5 spinal cord. Then we observed the influence of GABAa receptor antagonist (Bicuculline) on analgesic effect of CQ prescription.The results indicated that CQ prescription could remarkably increase MWT of model mice(P<0.01, P<0.05), decrease the level of Glu(P<0.01, P<0.05), improve the levels of GABA, Gly, Tau(P<0.01, P<0.05), lower the ratio of Glu/GABA(P<0.01, P<0.05), and reduce the levels of RANTES, MCP-3(P<0.05) in the L3-L5 spinal cord, and GABAa receptor antagonist significantly blocked the analgesic effect of CQ prescription at two time points(P<0.05).This study showed that CQ prescription had significant analgesic effect on CIIMIP model mice, and its mechanism was associated with regulating the balance between excitability amino acid(EAA) and inhibitory amino acid (IAA) transmitters in central nervous system, partially activating GABAa receptor, and reducing the release of RANTES and MCP-3 in the spinal cord.

The present study was designed to explore the influence of water extracts of Atractylodes lancea rhizomes on the toxicity and anti-inflammatory effects of triptolide (TP). A water extract was prepared from A. lancea rhizomes and co-administered with TP in C57BL/6 mice. The toxicity was assayed by determining serum biochemical parameters and visceral indexes and by liver histopathological analysis. The hepatic CYP3A expression levels were detected using Western blotting and RT-PCR methods. The data showed that the water extract of A. lancea rhizomes reduced triptolide-induced toxicity, probably by inducing the hepatic expression of CYP3A. The anti-inflammatory effects of TP were evaluated in mice using a xylene-induced ear edema test. By comparing ear edema inhibition rates, we found that the water extract could also increase the anti-inflammatory effects of TP. In conclusion, our results suggested that the water extract of A. lancea rhizomes, used in combination with TP, has a potential in reducing TP-induced toxicity and enhancing its anti-inflammatory effects.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Atractylodes ; chemistry ; Cytochrome P-450 Enzyme System ; genetics ; Diterpenes ; toxicity ; Edema ; chemically induced ; pathology ; Enzyme Induction ; drug effects ; Epoxy Compounds ; toxicity ; Gene Expression Regulation ; drug effects ; Herb-Drug Interactions ; Liver ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phenanthrenes ; toxicity ; Plant Extracts ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Water ; chemistry