Exploration of the Potential Mechanism of Calculus Bovis in Treatment of Primary Liver Cancer by Network Pharmacology.

AIM AND OBJECTIVE: Calculus Bovis (CB) has been employed to treat diseases for a long time. It has been identified to play significant anti-inflammatory and anti-tumor roles. However, the mechanism of treating primary liver cancer (PLC) remains to be revealed. This study aims to clarify the molecules and mechanisms of CB in treating PLC. MATERIALS AND METHODS: After oral bioavailability (OB) and drug-likeness (DL) screening, 15 small molecules were identified as the potential ingredients against PLC. Following this, related targets network constructions and pathways were applied to clarify the mechanism of CB in treating PLC. An in vitro experiment was carried out to identify the function of CB in treating PLC. RESULTS: Eleven compounds of CB were identified that play an anti-PLC role, including oleanolic acid, ergosterol, ursolic acid, etc. The potential targets which were observed include IL6, MAPK-8, VEGFA, Caspase-3, etc. Further analysis showed that the mechanism of CB in the treatment of PLC involved apoptosis-related pathways and immune-related pathways. CONCLUSION: In summary, the current study combines network pharmacology and in vitro experiments to reveal the mechanism of CB against PLC. We concluded that 11 ingredients of CB have an anti-PLC effect. Furthermore, CB plays a key role in treating PLC mainly by apoptosisrelated pathways and immune-related pathways. Our experiment verifies that CB promotes the apoptosis of SMMC-7721.

Oral Litholysis in Patients with Chronic Calcific Pancreatitis Unresponsive to or Ineligible for Extracorporeal Shock Wave Lithotripsy and Endoscopic Therapy.

BACKGROUND/AIMS: Our study aimed to evaluate the effect of oral litholysis in patients with chronic calcific pancreatitis (CCP) unresponsive to or ineligible for extracorporeal shock wave lithotripsy (ESWL) and endoscopic therapy. METHODS: Trimethadione, an antiepileptic agent, was administered orally at a dose of 0.6-0.9 g/day to 15 patients with this condition. Treatment outcome was evaluated by assessment of dissolution of the pancreatic stones on plain X-ray films and computed tomography scans of the upper abdomen. Plasma glucose, hemoglobin A1c, and body mass index (BMI) were also monitored throughout the therapy. RESULTS: Litholysis was observed in 13 out of 15 patients (86.7%) and was definite in 10 and partial in 3. Six patients had pancreatitis attacks during the therapy; 5 of whom showed definite litholysis and had only 1 mild attack. Of the 11 patients with normal or impaired glucose tolerance at baseline, none developed diabetes mellitus and all showed litholysis. BMI significantly increased in patients whose pancreatic stones dissolved. There was no vital organ impairment by trimethadione. CONCLUSION: Oral litholysis using trimethadione may represent a noninvasive and effective complementary treatment in patients with CCP unresponsive to or ineligible for ESWL and endoscopic therapy.

[Simultaneous determination of seven residual solvents in bovis calculus artifactus by headspace gas chromatography].

A headspace gas chromatography (HS-GC) method was developed for the simultaneous determination of seven residual solvents (petroleum ether (60-90 degrees C), acetone, ethyl acetate, methanol, methylene chloride, ethanol and butyl acetate) in bovis calculus artifactus. The DB-WAX capillary column and flame ionization detector (FID) were used for the separation and detection of the residual solvents, and the internal standard method was used for the quantification. The chromatographic conditions, such as equilibrium temperature and equilibrium time, were optimized. Under the optimized conditions, all of the seven residual solvents showed good linear relationships with good correlation coefficients (not less than 0.999 3) in the prescribed concentration range. At three spiked levels, the recoveries for the seven residual solvents were 94.7%-105.2% with the relative standard deviations (RSDs) less than 3.5%. The limits of detection (LODs) of the method were 0.43-5.23 mg/L, and the limits of quantification (LOQs) were 1.25-16.67 mg/L. The method is simple, rapid, sensitive and accurate, and is suitable for the simultaneous determination of the seven residual solvents in bovis calculus artifactus.

Development of fatty acid calcium stone ileus after initiation of human milk fortifier.

We report a case who was born with extremely low birth weight infant and had experienced abdominal operation for necrotizing enterocolitis, eventually developed ileus due to fatty acid calcium stones after giving human milk fortifier. He had developed necrotizing enterocolitis on day 30 of his age, such that we performed enterectomy and ileostomy. He could not tolerate enteral feeding fully, because intestinal fistula infection was repeated. Although we administered hindmilk, he grew up slowly and he suffered cholestasis as well. We performed end-to-end anastomosis to prevent fistula infections on day 87. After this operation, breast milk feeding volume was increased easily. However, we started to add HMF of half-strength on day 124, because his body weight gain remained very poor. And we confirmed to intensify the ratio of HMF full-strength on day 128. After that his abdomen had distended on day 131. As there is no effect of conservative therapy to occlusive ileus, we did emergency laparotomy on day 139. Intestinal calculi were impacted at anastomic portion. Although all stones were removed, he died on 144 days due to disseminated intravascular coagulation and renal failure. Calculi analysis revealed that all of them were fatty acid calcium stones. There is no report about like our case. We speculate that the construction of fatty acid calcium result from either high concentration of calcium/phosphorus or rapid increase in the fortification. We could have prevented this case happened by slower increment of fortification.

[Single and combining effects of Calculus Bovis and zolpidem on inhibitive neurotransmitter of rat striatum corpora].

OBJECTIVE: To investigate the correlation effects between single or combined administration of Calculus Bovis or zolpidem and changes of inhibitive neurotransmitter in rat striatum corpora. METHOD: Sampling from rat striatum corpora was carried out through microdialysis. The content of two inhibitive neurotransmitters in rat corpus striatum- glycine (Gly) and gama aminobutyric acid (GABA), was determined by HPLC, which involved pre-column derivation with orthophthaladehyde, reversed-phase gradient elution and fluorescence detection. RESULT: GABA content of rat striatum corpora in Calculus Bovis group was significantly increased compared with saline group (P < 0.01). GABA content of zolpidem group and Calculus Boris plus zolpidem group were increased largely compared with saline group as well (P < 0.05). GABA content of Calculus Bovis group was higher than combination group (P < 0.05). GABA content of zolpidem group was not significantly different from combination group. Gly content of Calculus Bovis or zolpidem group was markedly increased compared with saline group or combination group (P < 0.05). CONCLUSION: Contents of two inhibitive neurotransmitters in rat striatum corpora were all significantly increased in Calculus Bovis group, zolpidem group and combination group. The magnitude of increase was lower in combination group than in Calculus Bovis group and Zolpidem group, suggesting that Calculus Bovis promoted encephalon inhibition is more powerful than zolpidem. The increase in two inhibitive neurotransmitters did not show reinforcing effect in combination group, suggesting that Calculus Bovis and zolpidem may compete the same receptors. Therefore, combination of Calculus Bovis containing drugs and zolpidem has no clinical significance. Calculus Bovis shouldn't as an aperture-opening drugs be used for resuscitation therapy.

[A case of colonic obstruction due to post-operative stenosis of the colon and multiple enteroliths].

An 81-year-old man who had under gone two abdominal surgeries and temporary colostomy 30 years previously was admitted due to lower abdominal pain and vomiting. An abdominal X-ray film and abdominal CT scan showed intestinal distension and multiple calcareous deposits in the colon. Gastrografin enema examination revealed smooth stenosis at the sigmoid colon and many additional defects. Endoscopy could not be performed due to the stenosis. He did not agree to surgery. Seven months later, he was admitted again, due to colonic obstruction. Surgery was performed which revealed colonic obstruction as the source of post-operative stenosis of the sigmoid colon and multiple enteroliths. The stones consisted of a core and a hull and contained ammonium magnesium phosphate.

The protective effects of in vitro cultivated calculus bovis on the cerebral and myocardial cells in hypoxic mice.

The protective effects of in vitro cultivated calculus bovis (ICCB) on the cerebral and myocardial cells in hypoxic mice and the mechanism were examined. In one group, mice were intragastrically (i.g.) given ICCB for 15 days and then they were subjected to acute cerebral ischemia by decapitation, and then the panting time was recorded. In the other group, 12 min after exposure to hypoxia, mice was administered the ICCB i.g. for 5 days, and then the blood serum and tissues of brain, heart, liver were harvested and examined for SOD, GSH-px and T-AOC activity and content of MDA. The tissues of brain and heart were observed electron-microscopically for ultrastructural changes. The corpus striatum and hippocampus of brain were collected and examined for content of dopamine (DA) and norepinephrine (NE). The ultrastructural examination showed that the pathological change in brain and heart in the ICCB group was very slight, while abnormal changes in the control group were obviously more serious. ICCB significantly prolonged the panting time of the hypoxic mice (P<0.001), increased the activity of SOD, GSH-px, T-AOC in serum and tissues of brain, liver, heart and elevated the content of DA and NE. ICCB also pronouncedly reduced content of MDA in serum and tissues of brain, heart and liver. Significant differences in these parameters were noted between ICCB group and controls. It is concluded that ICCB can exert protective effect on the cells of brain and myocardium by enhancing the tolerance of the tissues to hypoxia and the body's ability to remove free radicals and regulating the neurotransmitters.

[Multiple enterolithiasis, coexisting with bladder and gallbladder lithiasis, associated with colon adenocarcinoma]. / Enterolitiasis múltiple, coexistiendo con litiasis biliar y vesical, asociada a adenocarcinoma de colon.

Enteroliths are calculi primarily formed in the intestine. Enterolithiasis is a rare condition frequently associated with intestinal stasis. Usually it causes no symptoms in most cases, but it can be an important diagnostic clue in patients presenting intestinal occlusive symptoms. We report a case of multiple enterolithiasis, very infrequent pathology, coexisting with bladder and gall bladder lithiasis in a patient with colon adenocarcinoma. Diagnosis was made by X-rays and CT images. Calculi were analysed by several methods: chemical, infrared spectroscopy, stereoscopic microscopy and atomic emission spectroscopy; they showed that caluli are made up of organic material and whilokita (calcium and magnesium ortophosphate). No risk factors for lithogenesis were found in this patient excluding the intestinal stasis caused by intestinal narrowing as a result of adenocarcinoma. Genetic factors are suggested as main contributors to hyperlithogenesis observed in this patient. The physiopathological conditions were studied in depth and literature about this subject reviewed.

In vitro CT evaluation of intrahepatic stones: correlation with chemical composition.

OBJECTIVE: To describe in vitro CT features of intrahepatic stones and to correlate CT attenuation with chemical composition. MATERIALS AND METHODS: Of the patients who underwent choledochoscopic intrahepatic stone removal between 1998 and 2001, 54 patients with stones larger than 3 mm were enrolled in this study. In each case, a chemical compositional analysis was performed to determine calcium, cholesterol, total bilirubin, and inorganic phosphorus compositions. The three largest stones obtained from each patient were imaged by CT. CT attenuation numbers were measured in the center images of each stone by drawing free-hand region of interest (ROI). The measured CT attenuation numbers were correlated with their chemical composition. Also, CT attenuation numbers of stones were compared with that of the liver on non-contrast CT (50-70 HU). RESULTS: Stone size ranged from 3.1 to 10.5 mm (mean +/- S.D.: 6.0 +/- 1.4). The CT attenuation numbers (HU) of stones ranged from 36.4 to 410.19 (mean +/- S.D.: 94.6 +/- 49.9). CT numbers of stones were below 70 HU in 11 patients (20.4%), and below 90 HU in 33 patients (59.3%). The chemical analysis data of the stones were as follows: calcium (0.5-6.5 wt.%; mean +/- S.D., 2.6 +/- 1.4), total bilirubin (0.45-24.4 wt.%; 13.1 +/- 6.2), cholesterol (5.4-73.9 wt.%; 29.3 +/- 17.4), phosphorus (0.1-1.2 wt.%; 0.6 +/- 0.3), and non-soluble residue (17.6-85.4 wt.%; 57.0 +/- 22.6). There was a weak but significant correlation between calcium composition and CT attenuation (r = 0.38, P < 0.01) and no significant correlation between other chemical compositions and the CT attenuation (cholesterol, r = 0.01, P > 0.01; total bilirubin, r = 0.05, P > 0.01; phosphorus, r = 0.01, P > 0.01). CONCLUSION: On non-contrast CT, intrahepatic stones would not be hyperattenuating with respect to liver parenchyma in about one fifth of patients. The CT attenuation of stones correlates with calcium and does not correlate with any other chemical composition.

[Prostatic calculi: diagnostic and therapeutic aspects]. / Osobennosti diagnostiki i lecheniia kamnei predstatel'noi zhelezy.

Strength properties of prostatic calculi (PC) were studied by microhardness HV. Fifteen calculi under study were divided into 3 groups by the size: 2-3 mm (group 1), 4-6 mm (group 2), > 6 mm (group 3). The surface of the thin sections of the calculi appeared as concentric, spherical layers of different size indicating formation of the calculus from the center. In the latter one could see a pigmented nucleus of organic nature. Some calculi had two nuclei under one cover--attached calculi. Correlations between the structure, morphology of prostatic calculi and their strength properties were established. These correlations allowed one to characterize destruction features of the calculi in different destructing impacts. We have designed and introduced into practice a complex of local physiotherapy in prostatolithiasis. This physiotherapy was conducted in combination with medicines for chronic prostatitis and included exposure of the prostate to ultrasound in impulse regime (4 mc, 0.4-1.0 W/cm2). The ultrasound was delivered through the rectum (insertion of the setting IUT 2.264-7-2.09u to the site of the concrements location) and alternated with laser therapy (insertion of the radiator P-2 into the rectum and radiator LO1 to the perineal area (5 W, 80 Hz, 64-128 s) and introduction of sulfide mud heated with the device PPK-01 Pra, electrostimulation for better passage of the prostatic section. The above therapy destructed the concrements which eliminated from the prostatic ducts as a result of which prostatic function recovered. The complex produced a good effect: 65% patients stated enhancement of the erectile function after 3 days of the treatment. After 5-6 procedures pain syndrome relieved in 40% patients, after 11-15 procedures in 87%. Ultrasonic investigation showed that posttreatment size of the prostate diminished from 0.8 to 0.2 cm. The absence of the calcinates's shadows were recorded in 87% patients, the symptoms of chronic prostatitis disappeared. The results of the combined physiotherapy demonstrate efficacy of the above method in true prostatoliths.

Stone fragility: its therapeutic implications in shock wave lithotripsy of upper urinary tract stones.

OBJECTIVE: To analyse the impact of stone composition on stone fragility (fragmentation) and clearance of upper urinary tract stones after shock wave lithotripsy (SWL). MATERIAL AND METHODS: Between 1st July 1998 and 31st July 2001, 300 renal and ureteric units of 290 patients (10 being bilateral) underwent SWL for upper urinary tract calculi. The degree of fragmentation was divided into four types: (I) Excellent, (II) Good, (III) Fair and (IV) No fragmentation. Stone composition was done by X-ray diffraction crystallography. A statistical comparison was made between degree of fragmentation, number of shock waves delivered, voltage setting, number of sessions required and requirements of adjuvant procedures according to the stone composition. RESULTS: Stone analysis revealed that 90% of the patients had calcium oxalate stones. Of these 80% were calcium oxalate monohydrate (COM) and 20% calcium oxalate dehydrate (COD). Struvite, apatite and uric acid stones comprised of 6%, 3% and 1% respectively. Type-I fragmentation was achieved up to 63.96%, 50% and 100% in COD, struvite and uric stones respectively as compared to 44.9% and 44.44% for COM and apatite stones. Type-III fragmentation was seen up to 8.79% and 33.3% respectively in COM and apatite as compared to 5.55% or less in other types of the stones suggesting that COM and apatite stones produce larger fragments. The mean number of shock waves, voltage and number of treatments was significantly higher for COM and apatite stones (p value < 0.005) with a stone free rate of only 65-66% and 65-68% respectively at three months (p value < 0.001). Similarly the number of adjuvant procedures required in COM alone was more, i.e. 31 as compared to 17 procedures in rest of the other kinds of stones (p value < 0.05). CONCLUSION: Stone composition in Indian subcontinent is different from the western world. Fragility of a stone varies with the composition of the stone and affects the therapeutic results.

Pulmonary alveolar microlithiasis. A clinicopathologic and chemical analysis of seven cases.

OBJECTIVE: To determine the clinical features and outcome of patients with pulmonary alveolar microlithiasis and to determine the chemical composition of the microliths. CASE MATERIAL: We studied seven cases of pulmonary alveolar microlithiasis. The patients were six women and one man, aged 19 to 70 years (mean age 44.5 years). Clinically, five patients were known to have suffered from this condition for 5 to 41 years. One patient presented with shortness of breath, and another had a gradual decrease in exercise tolerance. None of the patients had a previous history of disturbances in metabolism or any other relevant medical condition. Reports on radiographic studies were available in six cases, and chest radiographs were available for review in the seventh case. They all showed diffuse bilateral pulmonary infiltrates. Open lung biopsies were performed in two patients, and autopsy lung material was reviewed in five patients. RESULTS: Histologically, the lung showed the typical features of pulmonary alveolar microlithiasis, that is, presence of numerous microliths filling the alveolar spaces with either a normal or thickened fibrotic interstitium. Chemical analysis performed on the lung tissue of six of these patients revealed that the microliths consisted principally of calcium and phosphorus salts. Five of these patients died of respiratory failure; however, their deaths occurred from 5 to 41 years following the initial diagnosis. No follow-up information was obtained in two patients. CONCLUSIONS: The findings of this study confirm that pulmonary alveolar microlithiasis can be seen in any age group and that the microliths are composed principally of salts of calcium and phosphorus. Additionally, these cases confirm that the disease typically follows a protracted course.

Experimental dissolution of pigment gallstone material using alkaline EDTA and adjuvant bile salts/non-bile salt detergents, thiols and urea, with respect to local chemolitholysis.

In order to enhance the dissolution capacity and the kinetics of topical solvents used in local pigment chemolitholysis, a series of dissolution experiments was performed with intact brown and black pigment stones and with standardized solutes such as pigment stone powder and compressed powder (static disc method). The basic dissolution medium was a 0.1 M boric acid/sodium carbonate buffer (pH 9.5), and the basic lytic agent was EDTA-4Na, working satisfactorily at 1-3 g/dl. It could be demonstrated that the dissolution efficiency of this basic solvent was enhanced significantly in the presence of a detergent (surfacant) and of urea. Among the detergents the zwitterionic (e.g., Sulfobetain-12) and the nonionic types (e.g., a polyoxyethylene ether like Lubrol PX) proved to be most effective. The adjuvant effect of the investigated thiols was disappointing. Only dithioerythritol/dithiothreitol and N-acetylcysteine showed any moderate, if consistent, lytic activity. The highest dissolution rates in dissolving compressed powder standards (disc method) were achieved with the ternary solvent (1% w/v EDTA/80 mM Lubrol PX/1 M urea, pH 9.5). Intact black pigment stones, well known as problematic candidates for chemolitholysis, could be largely dissolved up to approximately 70% of their initial weight. This was not merely a physical disintegration, but a chemical process.

[Ultrastructure and elemental composition of pancreatic stones].

Four patients with pancreatic duct stones (PDS) and one with calcium deposits (CD) were diagnosed and surgically cured. The ultrastructure and elemental composition of PDS were studied by scanning electron microscopy and energy dispersive X-ray fluorescence. The results indicated that the outer shell of PDS, with dense texture, contained 96.29% of calcium (Ca), while the amorphous reticular inner nidus mainly contained sulphur (S) and chlorine (Cl). CD and PDS were manifestations in the process of pancreatic stones (PS) formation at different stages. As regards the formation of PS, the depressed gene expression played a key role in the reduction of PSP (Pancreatic Stone Protein), which made pancreatic acini reduce PSP biosynthesis rates. The consequence arising therefrom was that the supersaturated calcium carbonate (CaCO3) in pancreatic juice became inadequately inhibited. Thus the calcium carbonate became crystallized and deposited onto the reticular inner protein nidus (RIPN), which developed from the compilation of micro-protein-plugs that had grown from pathologically hyperconcentrated total proteins (mainly containing lactoferrin) in pancreatic juice. The RIPN gave a basis on which the calcium carbonate precipitated and formed PS.