OBJECTIVETo observe the effect of purgation and detoxification therapy on gastrointestinal dysfunction of critically ill patients undergoing abdominal surgery.

METHODSTotally 56 inpatients with severe gastrointestinal dysfunction after abdominal surgery at ICU of Guangdong Provincial Hospital of Traditional Chinese Medicine were assigned to the treatment group and the control group, 28 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took Modified Huanglian Jiedu Decoction (MHJD) and received electroacupuncture (EA) for 7 days. The first exhaust time, defecation time, scores for gastrointestinal dysfunction, mechanical ventilation time, ICU hospitalization time, and 28-day fatality rate were observed. Furthermore, serum levels of diamine oxidase (DAO) and D-lactic acid were detected at day 1, 3, and 7 after treatment.

RESULTSThe first exhaust time and the first defecation time in the treatment group were ahead of schedule, when compared with those of the control group (P <0. 05). Scores for gastrointestinal dysfunction, mechanical ventilation time, serum levels of DAO obviously decreased in the treatment group (P <0. 05). There was no statistical difference in serum levels of D-lactic acid, ICU stay time, the incidence of pulmonary infection, and 28-day mortality between the two groups (P >0. 05). Results of Logistic analysis showed that scores for gastrointestinal dysfunction were related with the incidence of pulmonary infection (P <0. 05).

CONCLUSIONMHJD combined EA could promote the recovery of gastrointestinal function in critically ill patients after abdominal surgery via improving intestinal barrier function, which was benefit for shortening mechanical ventilation time.

Critical Illness ; Defecation ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Electroacupuncture ; Gastrointestinal Diseases ; drug therapy ; Humans ; Lactic Acid ; Medicine, Chinese Traditional

OBJECTIVE Hypericin, a powerful naturally photosensitizer in photodynamic therapy (PDT), is suitable for treating skin diseases involving excess capillary proliferation. In the present study, we aimed to evaluate the skin penetrability of a topically applied hypericin, expecting reducing the risk of prolonged skin photosensitivity, which often occurs after systemic administration. METHODS The Franz diffusion cell assay was performed to evaluate different penetration enhancers. In vivo studies, fluorescence microscopy was performed to examine the distribution of hypericin in the skin, macroscopic and microscopic analyses were also carried out to detect pathological changes in the skin after topical hypericin-PDT treatment. Immunohistochemistry was used to determine the expression of PECAM-1 in the treated skin. RESULTS 5% menthol facilitated hypericin penetrate the skin of nude mice most. The results of in vivo assays revealed that hypericin penetrated nude mice skin, spread to the dermis, and resulted in obvious photosensitivity reaction on the dermal capillaries. Moreover, skin injured by the photosensitive reaction induced by hypericin was replaced by normal skin 7 d after hypericin-PDT treat?ment. CONCLUSION Topical hypericin could penetrate nude mouse skin well and be great potential in PDT treatment of skin diseases.

A cDNA fragment locating at the putative envelop protein 2(E2) region of GBV-C/HGV fused with Schistosoma japonicum, glutathione S-transferase(GST) was amplified with PCR from plasmid pGEX-E2. The amplified DNA fragment was inserted into plasmid pGEX-5X-1, at the downstream of the coding sequences of GST, in the same reading frame with the gene of GST. The fusion gene fragment of GST-E2 was amplified with PCR, using the recombinant plasmid pGEX-5X-1-E2 as the template. The amplified 1324 bp DNA fragment of GST-E2 was inserted into Pichia pastoris expression vector pPIC9K in reading frame with alpha-factor secreting signal peptide. The plasmid pPIC9K-GST-E2 was transformed into Pichia pastoris GS115 with electroporation. The transformants (His+ Muts) were selected and induced to express the 54kD GST-E2 fusion protein, which could be specially recognized by both the antisera directed against E2 and against GST. The GST-E2 fusion protein was purified with Sepharose 4B glutathione affinity chromatography to a purity of 95%. The expression was optimized to achieve the highest expression level of GST-E2 fusion protein which was accumulated up to 50% of total proteins in the culture supernatant. The GST-E2 protein derived from the recombinant Pichia pastoris was proved possessing antigenicity and high specificity by ELISA, probed with sera from the patients infected by GBV-C/HGV.
Animals ; Antigens, Viral ; genetics ; immunology ; isolation & purification ; GB virus C ; genetics ; immunology ; Gene Expression ; Genetic Engineering ; Glutathione Transferase ; genetics ; Hepatitis Antibodies ; blood ; immunology ; Humans ; Pichia ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification ; Schistosoma japonicum ; enzymology ; Viral Envelope Proteins ; genetics ; immunology ; isolation & purification

OBJECTIVETo study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA).

METHODRats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA.

RESULTCompared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA.

CONCLUSIONXLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

Animals ; Arthritis, Experimental ; immunology ; pathology ; prevention & control ; Cell Proliferation ; drug effects ; Colubridae ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Edema ; immunology ; pathology ; prevention & control ; Female ; Immunoglobulin G ; metabolism ; Lymphocytes ; drug effects ; pathology ; secretion ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Spleen ; pathology ; secretion ; Thymus Gland ; pathology ; Toes ; pathology

BACKGROUND: Intra-articular injection of cells targeting to change the microenvironment in lesions can act on early osteoarthritis of inflammatory chondrocytes. Implanted cells affect the progress of the disease by the cell characteristics. OBJECTIVE: To explore the synergistic effect of mesenchymal stem cells from human knee adipose (ADMSCs) and synovial tissues (SDMSCs) to inhibit the degeneration of inflammatory chondrocytes. METHODS: ADMSCs, SDMSCs and inflammatory chondrocytes were primary cultured. Under in vitro two-dimensional culture conditions, cell proliferation assay (MTS) was performed to detect the proliferation of three kinds of cells. Differences in chondrogenic markers at mRNA and protein levels between three kinds of adherent cells were detected by quantitative PCR and immunofluorescence. Under in vitro three-dimensional mixed culture conditions, three groups were set up: (1) ADMSCs+inflammatory chondrocytes (A+C group), (2) SDMSCs+inflammatory chondrocytes (S+C group), and (3) ADMSCs-SDMSCs+inflammatory chondrocytes (A+S+C group). Alcian blue staining, safranin O staining and type Ⅱ collagen immunohistochemistry staining were performed on the mixed-cultured cell mass paraffin sections followed by quantitative analysis. Chondrogenic differentiation in each group was detected by quantitative PCR. Culture supernatants were collected to detect the secretion of pro-inflammatory and anti-inflammatory factors by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: Under the two-dimensional culture, the proliferative rate of ADMSCs was significantly higher than that of inflammatory chondrocytes and SDMSCs (P < 0.05). The expression of type Ⅱ collagen mRNA and protein and proteoglycan protein in inflammatory chondrocytes was significantly higher than that in the other two kinds of cells (P < 0.01). Under the three-dimensional culture, the percentage of chondrogenic area per total area was significantly higher in the A+S+C group than the S+C and A+C groups (P < 0.05). The expression of type Ⅱ collagen and proteoglycan was significantly higher in the A+S+C group than the S+C and A+C groups (P < 0.05). Compared with the other two groups, the S+C group showed higher levels of interleukin 1, interleukin 6, and tumor necrosis factor α, but lower level of interleukin 10 (P < 0.05). To conclude, the combined use of ADMSCs and SDMSCs synergistically inhibits the degeneration of inflammatory chondrocytes.

This article summarized the clinical experience of Professor WANG Meng-yong in treating lupus nephritis from the symptoms and essence. Professor WANG Meng-yong believes that treatment for lupus nephritis should attach importance to clinical features of setting deficiency of liver, spleen and kidney as the essence and toxin and blood stasisas symptoms, in which the essence is deficient and the symptoms are excess and combination of external and internal pathogenesis. He flexibly uses methods of nourishing liver and kidney, tonifying spleen, and stomach, clearing heat and cooling blood, activating blood and promoting diuresis to alleviate water retention, which have obvious efficacy, effectively avoid adverse reactions and reduce the risk of recurrence, and improve patient prognosis.

Objective:To investigate the diagnostic and early-warning value of laboratory test indicators for sepsis-induced myocardial injury (SIMD).Methods:The clinical data of 183 patients with sepsis admitted to the Department of Emergency and Critical Care Medicine of Guangdong Provincial People's Hospital from August 2016 to October 2020 were collected. The patient's age, gender, past medical history, vital signs and pathogen culture results were extracted. Cardiac function, blood routine, liver function, renal function, inflammatory factors, coagulation function, APACHE Ⅱ and SOFA scores were recorded at enrollment and 72 h after admission. SIMD was defined as cTnT ≥300 pg/mL and NT-proBNP ≥1243 pg/mL twice in 72 h intervals between enrolled cases, and the early-warning factors of patients with SIMD were analyzed. The differences in various indicators between the two groups were compared, and Logistic regression analysis was used to explore the diagnostic efficacy of cTnT and NT-proBNP combined for SIMD, and the correlation between PCT/PLT ratio and the occurrence of SIMD.Results:Among 250 patients, 67 patients were excluded for lack of the main indicators, and 183 patients (including 62 patients with history of cardiac disease) were enrolled finally. Among 183 patients with sepsis, 105 patients (57.38%) with cTNT ≥300 pg/mL and NT-proBNP ≥1 243 pg/mL, were diagnosed as myocardial injury; after excluding 62 patients with history of cardiac disease, 59 patients (48.76%) with cTNT ≥300 pg/mL and NT-proBNP ≥1 243 pg/mL were diagnosed as myocardial injury. Logistic regression analysis showed that increased PCT/PLT ratio ( OR=1.585, 95% CI: 1.124-2.237, P=0.009) was an independent risk factor for early-warning of SIMD. The PCT/PLT ratio ( OR= 1.850, 95% CI: 1.103-3.102, P=0.020) could stably predict the occurrence of SIMD in patients without previous history of heart disease. ROC curve analysis showed that PCT/PLT ratio could effectively predict the occurrence of SIMD (AUC=0.693, 95% CI: 0.617-0.769, P<0.001), the optimal cut-off value was 0.177 (sensitivity: 65.7%, specificity: 66.7%). The PCT/PLT ratio was still effective in predicting the occurrence of SIMD after excluding patients with previous history of heart disease (AUC=0.733, 95% CI: 0.643-0.823, P<0.001), and the optimal cut-off value was 0.429 (sensitivity: 55.9%, specificity: 83.9%). Conclusions:The combination of cTnT and NT-proBNP has certain diagnostic value for SIMD, and the PCT/PLT ratio could warn the occurrence of SIMD.

Objective:To investigate whether stress hyperglycemia (SH) is an independent risk factor for the occurrence and mortality of sepsis-associated encephalopathy (SAE).Methods:From August 2016 to October 2021, sepsis patients admitted to the ICU of Guangdong Provincial People's Hospital were selected as the study subjects. According to whether they developed to SH (RBG>11.1 mmol/L) within 7 days of enrollment, the pat ients were divided into the SH group and the non-SH group for analysis. Logistic regression was used to analyze whether SH was an independent risk factor for SAE occurrence, and ROC curve was used to analyze the predictive value of SH to SAE. Kaplan-Meier curve was used to compare the 90-day survival of SAE patients with or without SH. Cox regression analysis was used to analyze the risk factors of 28-day and 90-day death in SAE patients.Results:A total of 183 sepsis patients were included, including 62 patients in the SH group and 121 in the non-SH group. Logistic regression analysis demonstrated that SH was an independent risk factor for SAE ( OR=4.452, 95% CI: 2.021-9.808, P <0.001). ROC curve demonstrated that SH could accurately predict SAE (AUC=0.831; Sensitivity=78.4%; Specificity=76.8%; and Yoden index=0.553). Kaplan-Meier curve demonstrated that the 90-day survival of SAE patients with SH significantly declined (log-rank test: P<0.01). Cox regression analysis suggested that SH was a risk factor for death at day 28 and day 90 in SAE patients (28 d, HR=2.272, 95% CI: 1.212-4.260, P=0.010; 90 d, HR=2.456, 95% CI: 1.400-4.306, P<0.01). Conclusions:SH is an independent risk factor for SAE and can predict SAE occurrence. SH significantly reduces 90-day survival and increase mortality at 28 and 90 days in SAE patients.

Objective To observe the changes of cervical sagittal parameters of digital radiography(DR)at bilateral oblique position of cervical spine in patients with neck type cervical spondylopathy(NTCS)after cervical rotation and lifting manipulations,and to evaluate the clinical significance of cervical sagittal parameters in assessing the efficacy of NTCS by combining the pain score of visual analogue scale(VAS).Methods A retrospective analysis was carried out in 36 NTCS patients admitted to the outpatient clinic of the Tuina Department of Foshan Hospital of Traditional Chinese Medicine from November 2020 to May 2021.According to the treatment method,the patients were divided into the trial group and the control group,with 18 patients in each group.The trial group was treated with cervical rotation and lifting manipulations,and the control group was treated with cervical vertebra traction.The two groups were treated once every other day for 2 continuous weeks.The changes of VAS pain score and sagittal parameters of DR such as sagittal vertical axis of cervical vertebra 2-7(C2-7 SVA),the T1 slope(T1S),neck tilt(NT),and thoracic inlet angle(TIA)in the two groups were observed before and after treatment.Spearman correlation analysis was used to explore the correlation between VAS scores of pain level and cervical sagittal parameters.Results(1)Before treatment,there was no statistically significant difference in the VAS pain scores between the two groups(P＞0.05).After treatment,the VAS pain scores in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the decrease of the scores in the trial group was significantly superior to that in the control group(P＜0.01).(2)Before treatment,there was no statistically significant difference in the values of cervical sagittal parameters of C2-7 SVA,T1S,NT,and TIA between the two groups(P＞0.05).After treatment,the C2-7 SVA,T1S,and TIA of the trial group were significantly improved compared with those before treatment(P＜0.05),but the improvement of NT was not obvious(P＞0.05);the cervical sagittal parameters of the control group were not significantly improved compared with those before treatment(P＞0.05).The intergroup comparison showed that the improvement of C2-7 SVA,T1S,and TIA in the trial group was significantly superior to that in the control group(P＜0.05 or P＜0.01),while the difference in NT between the two groups after treatment was not statistically significant(P＞0.05).(3)The results of Spearman correlation analysis showed that before treatment,VAS pain score had no correlation with C2-7 SVA,T1S,NT,and TIA(P＞0.05),C2-7 SVA was negatively correlated with NT(r=-0.502,P＜0.05),and T1S was positively correlated with NT(r=0.601,P＜0.05).After treatment,VAS pain score was negatively correlated with C2-7 SVA(r=-0.362,P＜0.05)and positively correlated with TIA(r=0.476,P＜0.05),C2-7 SVA was positively correlated with NT(r=0.928,P＜0.05),and T1S was positively correlated with TIA(r=0.623,P＜0.05).Conclusion Both cervical rotation and lifting manipulations and cervical traction can relieve the pain and adjust cervical sagittal parameters in the patients with NTCS,but cervical rotation and lifting manipulations are more effective in treating NTCS.And there may be a correlation between the VAS scores of pain level and cervical sagittal parameters in patients with NTCS.

Objective To investigate the osteogenic properties of a methacrylated gelatin(GelMA)/bone marrow mesenchymal stem cells(BMSCs)composite hydrogel applied to the skull defect area of rats and to provide an experi-mental basis for the development of bone regeneration biomaterials.Methods This study was approved by the Animal Ethics Committee of Nanjing University.A novel photocurable composite biohydrogel was developed by constructing photoinitiators[lthium phenyl(2,4,6-trimethylbenzoyl)phosphinate,LAP],GelMA,and BMSCs.The surface morphology and elemental composition of the gel were examined using scanning electron microscopy(SEM)and energy-dispersive X-ray spectroscopy(EDX).The compressive strength of the gel was evaluated using an electronic universal testing ma-chine.After in vitro culture for 1,2,and 5 days,the proliferation of the BMSCs in the hydrogels was assessed using a CCK-8 assay,and their survival and morphology were examined through confocal microscopy.A 5 mm critical bone de-ficiency model was generated in a rat skull.The group receiving composite hydrogel treatment was referred to as the Gel-MA/BMSCs group,whereas the untreated group served as the control group.At the 4th and 8th weeks,micro-CT scans were taken to measure the bone defect area and new bone index,while at the 8th week,skull samples from the defect ar-ea were subjected to H&E staining,van Gieson staining,and Goldner staining to evaluate the quality of bone regenera-tion and new bone formation.Results SEM observed that the solidified GelMA showed a 3D spongy gel network with uniform morphology,the porosity of GelMA was 73.41％and the pore size of GelMA was(28.75±7.13)μm.EDX results showed that C and O were evenly distributed in the network macroporous structure of hydrogel.The hydrogel compres-sion strength was 152 kPa.On the 5th day of GelMA/BMSCs culture,the cellular morphology transitioned from oval to spindle shaped under microscopic observation,accompanied by a significant increase in cell proliferation(159.4％,as determined by the CCK-8 assay).At 4 weeks after surgery,a 3D reconstructed micro-CT image revealed a minimal re-duction in bone defect size within the control group and abundant new bone formation in the GelMA/BMSCs group.At 8 weeks after surgery,no significant changes were observed in the control group's bone defect area,with only limited evi-dence of new bone growth;however,substantial healing of skull defects was evident in the GelMA/BMSCs group.Quan-titative analysis at both the 4-and 8-week examinations indicated significant improvements in the new bone volume(BV),new bone volume/total bone volume(BV/TV),bone surface(BS),and bone surface/total bone volume(BS/TV)in the GelMA/BMSCs group compared to those in the control group(P＜0.05).Histological staining showed continuous and dense formation of bone tissue within the defects in the GelMA/BMSCs group and only sporadic formation of new bone,primarily consisting of fibrous connective tissue,at the defect edge in the control group.Conclusion Photocur-ing hydrogel-based stem cell therapy exhibits favorable biosafety profiles and has potential for clinical application by inducing new bone formation and promoting maturation within rat skull defects.